Publisher(s): China Academic Journals (CD Edition) Electronic Publishing House Co., Ltd.
ISBN: ISBN 978-7-499-00978-3 pdf
First Published: 2020.11.23
Discipline(s): Medicine & Public Health
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Inflammation, as part of China’s Medicine Progress Series, has 76 excellent articles on inflammation research. These articles included were recommended by the editorial boards of the journals in which they were originally published, covering theoretical, drug, mechanism, clinical, and nursing research and fully demonstrating the cutting-edge advances in inflammation treatment in China, with methodological support and objective conclusions. The original articles are published in Chinese, and this book is a compilation of English versions of selected articles.
LIU Changxiao is the honorary president, tenured principal scientist and academic committee director of Tianjin Institute of Pharmaceutical Research, and the director of the State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, the Center
1. Experiment of THP-1 macrophage foam-cell formation through mTOR signal pathway activation induced by inflammatory stress
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 32,No. 01
Aim To investigate if LPS increases the sterol regulatory element binding proteins (SREBPs) cleavage-activating protein (SCAP)-SREBP2 expression by activation of mTOR signal pathway in THP-1 macrophages, upgrading LDLr level, causing foam-cell formation. Methods THP-1 macrophages were incubated in serum-free medium with 5 mg·L – 1 LDL alone, or 5 mg·L – 1 LDL plus 200 μg·L – 1 LPS, or 5 mg·L – 1 LDL plus 200 μg·L – 1 LPS plus 10 μg·L – 1 rapamycin. Morphological examination of macrophages was performed with Oil Red O staining. Expression changes of LDLr, SREBP2, SCAP, S6K1 and mTOR mRNA were detected by real-time quantitative polymerase chain reaction (PCR). Western blot was used to analyze protein expression changes of LDLr, S6K1 and mTOR. Translocation of SCAP-SREBP2 complex from the endoplasmic reticulum (ER) to the Golgi was determined by confocal microscopy. Results LPS enhanced transformation of THP-1 macrophages into foam cells by increased uptake of lipid as evidenced by Oil Red O assay. LPS increased mRNA levels of LDLr, SREBP2, SCAP, S6K1 and mTOR ( P < 0.05). Rapamycin reduced the mRNA levels of LDLr, SREBP2, SCAP, S6K1 and mTOR induced by LPS ( P < 0.05). Western blot demonstrated that LPS also caused over-expression of protein of LDLr, S6K1 and mTOR ( P < 0.05). Rapamycin reduced the expression of protein of LDLr, S6K1 and mTOR induced by LPS ( P < 0.05). Confocal microscopy demonstrated LPS caused an escape of SCAP-SREBP2 complex from the ER to the Golgi. Rapamycin inhibited the translocation of SCAP-SREBP2 complex from the ER to the Golgi. Conclusions Inflammatory stress increases SCAP/SREBP2 expression by activation of mTOR signal pathway, resulting in an escape of SCAP-SREBP2 complex from the ER to the Golgi, furthermore elevating LDLr expression and causing foam-cell formation. Rapamycin reverses the activation of mTOR signal pathway and decreases lipid deposition in THP-1 macrophages induced by LPS.
2. Inflammatory mechanism of acute lung injury in mice induced by activation of complement alternative pathway
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 32,No. 02
Aim To study the development of acute lung inflammation in mice induced by activation of the complement alternative pathway and the changes of the related indicators, and to provide an ideal pathologicalmodel of acute lung inflammation in mice for drug screening and intervention. Methods Cobra venom factor (CVF) was used to activate complement alternative pathway of SPF Kunming mice by intravenous injection. According to different sampling time, the mice were divided into 15 min, 30 min, 1 h, 2 h, 6 h groups, and the parallel PBS control groups were set at the same time. Lung coefficient, lung water content, myeloperoxidase (MPO) activity, BALF cell number and protein content were tested. The pathological changes of lung tissue were observed by HE staining. The concentration of IL-6, TNF-α, P-selectin and ICAM-1 in bronchoalveolar lavage fluid (BALF) and serum were determined by ELISA. Results CVF caused pulmonary inflammatory cell infiltration in mice obviously. Compared with PBS groups, MPO activity of lung tissue, BALF cell and the protein concentration were significantly increased. The contents of IL-6, TNF-α, P-selectin in BALF and serum were increased, and the content of ICAM-1 in serum was also increased. The content of P-selectin in BALF reached the first peak at 30 min point, the contents of IL-6 and TNF-α in BALF reached the first peak at 1 h point, but the indicators had no further changes at 2 h point, and all the indicators rose again at 6 h point. The levels of IL-6 and TNF-α in serum reached peak at 1 h point, then the content showed lower levels at the subsequent time points. The levels of P-selectin and ICAM-1 in serum increased along the time. Lung coefficient, lung water content and ICAM-1 of the BALF showed no significant alteration. Conclusion The activation of the complement alternative pathway could lead to acute lung inflammation in mice and the inflammatory response was the most obvious at 30 min to 1 h. The study could provide an ideal pathological model of acute lung inflammation in mice for drug screening and intervention.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 32,No. 03
Aim To investigate whether nicorandil (Nic) protects H9c2 cardiac cells against high glucose ( HG)-induced injury and inflammation by inhibiting the nuclear factor-κB (NF-κB) / cyclooxygenase-2 ( COX-2) pathway. Methods The cell viability was measured by cell counter kit-8 (CCK-8) assay. The expression levels of NF-κB, COX-2 and cleaved caspase-3 were determined by Western blot. The activity of lactate dehydrogenase (LDH) in the culture medium was measured with commercial kits. The intracellular level of reactive oxygen species (ROS) was detected by 2',7'-dichlor-fluorescein-diacetate (DCFH-DA) staining followed by photofluorography. The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography. The mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorography. The secretion levels of interleukin ?1β (IL ?1β) and tumor necrosis factor-α (TNF-α) were detected by ELISA. Results After the H9c2 cardiac cells were treated with 35 mmol·L ?1 glucose (HG) for 24 h, the cell viability was significantly decreased. Pre-treatment of the cells with 20–100 μmol·L ?1 Nic for 60 min or 50 μmol·L ?1 Nic for 30–120 min before exposure to HG significantly attenuated the decrease in viability induced by HG. On the other hand, HG increased the expression levels of phosphorated (p)-NF-κB p65 and cyclooxygenase-2 (COX-2) in H9c2 cardiac cells. Pre-treatment of the cells with 50 μmol·L ?1 Nic for 60 min attenuated the up-regulation of p-NF-κB p65 and COX-2 expression levels induced by HG. Furthermore, HG induced considerable injuries and inflammatory response, leading to increases in the LDH activity, ROS generation, MMP loss, the number of apoptotic cells, the expression of cleaved caspase-3 as well as the secretion levels of IL ?1β and TNF-α. Pre-treatment of the cells with 50 μmol·L ?1 Nic for 60 min before HG exposure, or co-treatment of the cells with 100 μmol·L ?1 PDTC (an inhibitor of NF-κB) or 10 μmol·L ?1 NS-398 (an inhibitor of COX-2) and HG for 24 h obviously reduced the above injuries and inflammatory response induced by HG. Conclusion Nic protects H9c2 cardiac cells against HG-induced injury and inflammation by inhibiting the NF-κB / COX-2 pathway.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 33,No. 04
Aim To investigate whether necroptosis mediates chemical hypoxia-induced HT22 mouse hippocampal cell injury and inflammation. Methods HT22 hippocampal cells were exposed to cobalt chloride (CoCl 2) to establish a model of the chemical hypoxia-induced injury and inflammation. The expression level of RIP3 (an index of necroptosis) was determined by Western blot. Cell counter kit-8 (CCK-8) assay was used to test the cell viability. Lactate dehydrogenase (LDH) activity in the culture medium was measured with commercial kits. Mitochondrial membrane potential (MMP) was examined by rhodamine123 staining followed by photofluorography. The intracellular level of reactive oxygen species (ROS) was detected by 2’,7’-dichlorfluorescein-diacetate (DCFH-DA) staining followed by photofluorography. The secretion levels of interleukin-1β (IL-1β) and tumor necrosis factor-a (TNF-α) were measured by ELISA. Results Treatment of HT22 hippocampa cells with 600 μmol·L ?1 Co Cl 2 for 36 h markedly induced cytotoxicity, leading to a decrease in cell viability to (52.0 ± 2.65) %, indicating that chemical hypoxia-induced cellular injury model was successfully set up. Besides, CoCl 2 induced considerable injuries and inflammation, evidenced by increases in LDH activity, ROS production, MMP loss, as well as the secretion levels of IL-1β and TNF-α. Co-treatment of the cells with 40–100 μmol·L ?1 Nec-1 (a specific inhibitor of necroptosis) and CoCl 2 markedly attenuated the decrease in viability induced by CoCl 2, reaching the best anti-cytotoxicity inhibitory effect at 80 μmol·L ?1. Meanwhile, the co-treatment with 80 μmol·L –1 Nec-1 blocked the above injuries and inflammatory response induced by CoCl 2. In addition, treatment of HT22 hippocampal cells for 6–48 h up-regulated the expression of RIP3, and Nec-1 alleviated the up-regulation of RIP3 expression level induced by CoCl 2. Conclusion Necroptosis mediates chemical hypoxia-induced HT22 hippocampal cell injury and inflammation.
Chinese Pharmaceutical Journal,Part 3: Mechanism Study,Vol 52,No. 05
OBJECTIVE To establish dose-related lung inflammatory injury in rats model with intratracheal atomization of lipopolysaccharides (LPS). METHODS Four groups of rats were subjected to solvent or a single dose of LPS by intratracheal route using a IA-1B-2 inches-microsprayer. The male rats received 200 μL solvent (control), LPS solutions (15, 5, 0.5 mg·kg ?1). All rats were sacrificed 24 h after dose administration, biochemical analysis and cell counts on bronchoalveolar lavage fluid (BALF) were performed on each rat. Lung, trachea and kidney were examined histologically. Serum chemistry profiles of creatinine, ALB, Na +, K +, Cl ? were detected. RESULTS Cell counts in BALF showed LPS groups had different degrees of inflammatory reaction. The alkaline phosphatase and total protein concentration were higher in LPS high dose group compared with other groups. In addition, the concentration of TNF-α increased consistently with LPS dose and has statistical significance compared with the control group. Histopathology findings demonstrated that LPS produced an accumulation of foamy macrophages in the lungs and high degree of inflammation. CONCLUSION The results recommend intratracheally atomizing doses of LPS in rats model produced ranks of lung inflammatory injury.
6. Inhibition of the Inflammatory Response and Its Mechanism by Hydrochloric Acid Chlorobenzene Sulperzon (HACS) in Astrocytes
Chinese Pharmaceutical Journal,Part 3: Mechanism Study,Vol 52,No. 06
OBJECTIVE To evaluate the inhibition effects and mechanism of hydrochloric acid chlorobenzene sulperzon (HACS) on neuronal inflammation were studied in order to evaluate possible application of it in AD therapy. METHODS The release of NO (nitric oxide) by astrocyte was detected by Griess methods and the chemotaxis of mouse macrophage was detected by Boyden chemotaxis chamber. The cell viability was detected by MTT assay. The contents of IL-6 and RANTES (T cell expressed and presumably secreted) were determined by ELISA. The expression of mRNA of IL-6 and RANTES was detected by RT-PCR. The intracellular Ca 2+ was detected by confocal microscope. RESULTS HACS efficiently decreased the release of NO from astrocyte stimulated by LPS (1 μg·mL ?1), chemotaxis of mouse macrophage stimulated by PAF (5 × 10 ?8 mol·L ?1), expression of IL-6 and regulated upon activation of RANTES in U251 cells induced by IL-1β (50 ng·mL ?1). In addition, HACS significantly inhibited the increase of intracellular Ca 2 + in U251 cells induced by Aβ1-42 (50 μg·mL ?1) / sodium glutamate (100 μmol·L ?1) or IL-1β (50 ng·mL ?1). CONCLUSION HACS efficiently inhibits the activation of astrocyte by regulation of intracellular Ca 2 + inhibition of chemotaxis and decrease of inflammatory cytokines. Especially the inhibition of RANTES and intracellular Ca 2+ induced by inflammatory mediators by HACS was firstly reported in this study.
7. Effect of extracellular cyclophilin A on inflammatory response and anti-inflammatory activity of antibody against cyclophilin A
Chinese Journal of Biotechnology,Part 3: Mechanism Study,Vol 34,No. 07
Cyclophilin A (Cyp A) is a member of peptidyl prolylisomerases (PPIase) family. Cyp A is best known as a ubiquitously distributed intracellular protein. It has also been shown to be secreted by cells in response to inflammatory stimuli and oxidative stress. Extracellular Cyp A (eCypA) interacts with CD147 to initiate inflammatory responses via recruiting leucocytes into inflamed tissue. Recombinant Cyp A was expressed in Escherichia coli and then purified using Superdex 75 TM16/60. The results of Real-time PCR and ELISA showed that the expression levels of proinflammatory cytokines, such as IL-1β, secreted by eCypA stimulated BMDM were significantly up-regulated, indicating that eCypA played an important role in promoting inflammatory responses. In addition, anti-Cyp A antibody was prepared using purified Cyp A protein for therapeutic evaluation in a mouse model of LPS-induced acute lung inflammation. Antibody-treated mice showed reduced lung injury and the expression levels of IL-1βin the lung tissue and blood were decreased significantly, indicating that anti-Cyp A antibody exerted a potent anti-inflammatory activity. Our findings provide a potential therapeutic antibody for inflammation-mediated diseases.
8. Role of adenovirus-mediated cardiac-specific RIP140 overexpression in cardiac function and inflammation pathway
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 33,No. 08
Aim To explore the role of cardiac-specific overexpression of RIP140 in cardiac function and inflammation signaling pathway. Methods Direct intra-myocardial injection of adenovirus vector expressing RIP140 drove transgene expression in heart tissue. RIP140 overexpression was confirmed using immunofluorescence technique in heart. Cardiac function was assessed by echocardiographic and hemodynamic assessment. TNF-α, IL-2 and IL-1β inflammatory cytokines were detected by ELISA, and the protein levels of p65 and IκB-α were measured by Western blot. Results Adenovirus-mediated foreign gene of RIP140 was successfully transferred in cardiac tissue. The RIP140 overexpression in heart induced ventricular dilation, decreased left ventricular ejection fraction and cardiac malfunction, as well as increased releases of TNF-α, IL-2 and IL-1β inflammatory cytokines, p65 protein translocation into the nucleus and IκB-α protein degradation in cytoplasm. Conclusion The Adenovirus-mediated RIP140 overexpression in cardiac tissue impairs cardiac function, activates NF-κB/p65 inflammatory signaling pathway and induces the release of inflammatory cytokines.
9. Effect of FABP4 on placental trophoblastic immune inflammatory response in patients of hypertensive disorder complicating pregnancy
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 33,No. 09
Aim To investigate the role of fatty acid binding protein 4 (FABP4) in placental trophoblastic immune response in patients with hypertensive disorder complicating pregnancy. Methods The human placental trophoblast cell line (HTR-8) was cultured in vitro and incubated with L-NAME for 0, 10, 100, 500 and 1 000 μmol·L ?1for 48 h. The levels of IL-6 and TNF-α were analyzed by ELISA. The mRNA and protein expression levels of FABP4 were detected by Western blot and qRT-PCR. The cells transfected FABP4adenovirus expression vector were intervented with 100 μmol·L ?1L-NAME. Then, the levels of IL-6 and TNF-α were determined by ELISA. Results Compared with the control group, the expression levels of IL-6, TNF-α and FABP4 were significantly increased in the intervention group ( P < 0.01). Meanwhile, the expression levels of IL-6 and TNF-αwere significantly increased in the overexpressed FABP4 group, compared with the control group ( P < 0.01). Conclusion FABP4 is involved in regulating the trophoblastic immune-inflammatory response in patients of hypertensive disorder complicating pregnancy.
10. Effect of enalapril folic acid tablet on peripheral blood inflammatory factors in type H hypertensive rats
The Chinese Journal of Clinical Pharmacology,Part 3: Mechanism Study,Vol 33,No. 10
Objective To study the effect of enalapril folic acid on peripheral blood inflammatory factors in rats with type H hypertension. Methods Sixty Wistar Spontaneous hypertension male rats were randomly divided into control group, model group, enalapril group and enalapril folic acid group, with 15 rats in each group. Type H hypertensive rat model were established by administration with diet containing 3% methionine in the model group, enalapril group and enalapril folic acid group. And rats in the enalapril folic acid group were given 10 mL·kg ?1 enalapril maleate folic acid solution, those in the enalapril group were given 10 mL·kg ?1 enalapril solution, and those in the model and control groups were given 10 mL·kg ?1 double distilled water. The treatment was performed once per day, for eight successive weeks in total. The serum levels of homocysteine (Hcy), serum tumor necrosis factor alpha (TNF-α), hypersensitivity C reactive protein (hs-CRP) and soluble OX40 ligand (OX40L) were measured by enzyme-linked immunosorbent assay. The nitric oxide (NO) level were measured by End-point assay. The structure of carotid artery was observed under microscope. Results Compared with Hcy (64.22 ± 6.82) μmol·L ?1 and NO (41.45 ± 5.25) μmol·L ?1 in the enalapril group, levels of Hcy (19.75 ± 2.14) μmol·L ?1 were lower, and NO (57.68 ± 6.43) μmol·L ?1 were higher in the enalapril folic acid group, with significant differences ( P < 0.05). Compared with TNF-α (72.72 ± 6.89) pg·mL ?1, hs-CRP (1.15 ± 0.12) pg·mL ?1, and OX40L (5.24 ± 0.47) ng·mL ?1 in the enalapril group, levels of TNF-α (58.18 ± 5.76) pg·mL ?1, hs-CRP (0.86 ± 0.11) pg·mL ?1, and OX40L (3.03 ± 0.39) ng·mL ?1 were lower in the enalapril folic acid group, with significant differences ( P < 0.05). Compared with the IMT (61.14 ± 7.73) μm, and IMCSA levels (0.12 ± 0.02) mm 2 in the carotid artery of rats in the enalapril group, the IMT (52.74 ± 6.29) μm, and IMCSA levels (0.10 ± 0.01) mm 2 in the enalapril folic acid group were lower, with significant differences ( P < 0.05). Compared with the level of LD (951.38 ± 6.08) μm in the enalapril group, the levels of LD (969.79 ± 6.17) μm in the enalapril folic acid group was significantly higher ( P < 0.05). Conclusion Enalapril folic acid can effectively reduce levels of peripheral blood Hcy, TNF-α, hs-CRP and OX40 L in type H hypertensive rats, improve levels of NO, reduce inflammation, and improve the structure of carotid artery.
11. Formyl peptide receptor-2 enhances inflammatory response induced by lipopolysaccharide in BV-2 cells
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 34,No. 11
Aim To investigate the expression of formyl peptide receptor-2 (FPR2) in lipopolysaccharide (LPS)-induced-BV-2 cells, and detect FPR2’s influence on inflammatory response induced by LPS. Methods After 1 mg·L ?1 LPS acting on BV-2 cells at 12 h, the extrinsic inflammatory model was established. We used the Western blot assay to test the levels of FPR2 protein. And the expression levels of phosphorylated NF-κB, TNF-α and IL-1β were investigated when the LPS-induced-BV-2 was incubated with FPR2’s agonist MMK-1 and antagonist Boc-2. Transwell assay was also used to detect the LPS-induced BV-2 migration induced by MMK-1 and Boc-2. Results LPS up-regulated the expression of FPR2, and when its agonist acted on LPS-induced-BV-2, the levels of phosphorylated NF-κB, TNF-α and IL-1β were significantly higher than those of LPS group. In addition, the chemotaxis of LPS-induced-BV-2 was also increased by MMK-1. These effects were abolished by Boc-2. Conclusions LPS can increase the expression of FPR2 on BV-2 cells, and FPR2 enhances the inflammatory response induced by LPS.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 34,No. 12
Aim To investigate the effects of essential oil of Fructus Alpiniae zerumbet (EOFAZ) on angiogenesis and inflammatory injury induced by tumor necrosis factor-α (TNF-α) and the underlying mechanisms. Methods Human umbilical vein endothelial cells (HUVECs) angiogenesis model in vitro and mice thoracic aorta inflammatory injury model in vivo were established by TNF-α, respectively. Angiogenesis assay was used to investigate the effects of EOFAZ on the tube formation of HUVECs angiogenesis. Western blot and qRT-PCR were used to detect the protein and mRNA expressions of VEGFR-2 in HUVECs, respectively. The secretion level of VEGFR-2 in serum of mice was detected by ELISA, and HE staining assay was performed on inflammatory injury in thoracic aorta of mice. Results EOFAZ and ERK inhibitor U0126 significantly inhibited the tube formation of HUVECs angiogenesis ( P < 0.01), EOFAZ down-regulated the secretion level of VEGFR-2 in TNF-α-induced mice ( P < 0.01), EOFAZ and U0126 suppressed the protein and mRNA expression of VEGFR-2 in TNF-α-induced HUVECs ( P < 0.01), and EOFAZ inhibited inflammatory cells infiltration in thoracic aorta of TNF-α-induced mice. Conclusions EOFAZ significantly inhibits angiogenesis and inflammatory injury induced by TNF-α, the mechanism of the inhibitory effect of EOFAZ may be related to down-regulating the expression of VEGFR-2 via ERK signaling pathway.
13. Alpha lipoic acid provides protection on renal tissue fibrosis of diabetic rats by inhibiting activation of TLR4 and NLRP3 inflammatory signal
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 35,No. 13
Aim To investigate the effects of alpha lipoic acid (ALA) on Notch2, TLR4, NLRP3 and inflammatory factor expression in renal tissues of diabetes mellitus (DM) rats, and to explore its possible mechanism of anti-inflammatory and anti-fibrosis by Alpha lipoic acid. Methods The diabetic rat model was established by streptozotocin. The rats were divided into two groups: the DM group and ALA group. Meanwhile, and another eight rats were used as normal control (NC group). After eight weeks, the rats were sacrificed to detect the relevant biochemical parameters and oxidative stress indexes. In addition, immunohistochemical staining was employed to detect the protein expression localization sites of Notch2, TLR4 and NLRP3. Western blot analysis was used to detect the expression of Notch2, TLR4, NLRP3 and collagen Ⅳ proteins in renal tissues. ELISA was utilized to detect the inflammatory factor expression of IL-6 and TNF-α. Results Compared with NC group, the levels of blood glucose, 24 h urine protein, total cholesterol and triglyceride all increased in DM group, and the activity of T-AOC was reduced whereas the MDA content was up-regulated in DM group, all items but blood glucose were significantly reduced in ALA group, and the activity of T-AOC remarkably increased, while the MDA content was reduced in ALA group. Compared with NC group, the protein levels of Notch2, TLR4, NLRP3 and collagen Ⅳ in kidneys were raised, and the inflammatory factor expression of IL-6 and TNF-α significantly increased in DM group. Conclusions ALA may down-regulate the inflammatory signal of TLR4 and NLRP3 in kidney of diabetic rats, and reduce the expression of inflammatory factor and the accumulation of extracellular matrix via inhibiting the expression of Notch2 at protein level.
14. COX-2 pathway changes in hippocampus and cortex of rat offsprings following maternal inflammation during pregnancy
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 35,No. 14
Aim To observe the COX-2 pathway changes in hippocampus and cortex of rat offsprings following maternal inflammation during pregnancy. Methods Female SD rats were randomly divided into control group and lipopolysaccharide (LPS) group. The rats in LPS group were intraperitoneally injected with LPS 300 μg·kg ?1 on 11th, 14th and 18th day of pregnancy, while those in control group were given normal saline. The body weights of offspring rats on 3th, 10th, 20th and 30th day were recorded; on 30th day, the development of nervous system of the offspring rats was tested using the water maze test, open field test and spontaneous activity test and so on; the histopathological changes of the hippocampus and cortex were detected by hematoxylineosin staining; the levels of inflammatory factors were measured by ELISA; the protein expression of COX-2 was measured by Western blot. Results There were no significant differences in the body weight of offspring rats between NS group and LPS group ( P > 0.05). In LPS group, it was found that the learning and memory function of rats was impaired, and the horizontal movement and spontaneous activities were significantly reduced ( P < 0.05); the hippocampus and cortex neurons showed a significant nuclear pyknosis ( P < 0.05, P < 0.01); the levels of PGD2, PGE2, PGI2, TNF-α, IL-6 and IL-1β in hippocampus and cortex significantly increased ( P < 0.05, P < 0.01); the protein expression of COX-2 in hippocampus and cortex significantly increased ( P < 0.01). Conclusions COX-2 and the downstream pathway are involved in the mechanism of brain injury in offsprings of pregnancy inflammation rats.
15. Effect of a PI3K Inhibitor on NO Production and Inflammatory Response in the Lung Tissues of Mice with Viral Pneumonia Induced by Influenza A Virus FM1 Infection
Chinese Journal of Virology,Part 3: Mechanism Study,Vol 35,No. 15
Influenza A virus (IAV) infection can trigger cytokine storms and increase the risk of acute respiratory distress syndrome (ARDS). Cascade amplification of inflammatory reactions in lung tissue plays a key role in this process. Phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signalling pathway participates in the regulation of cellular inflammatory response, stimulates the expression of endothelial nitric oxide synthase (eNOS), and increases the production of nitric oxide (NO). Through the activity of NO, the PI3K/AKT signalling pathway can increase vascular permeability, promote the infiltration of inflammatory cells, stimulate the activation of inflammatory cells, and secrete a large number of inflammatory factors. However, the role of PI3K/AKT signalling pathway in the virulent pneumonia caused by IAV infection has not been clarified. To study the effects of PI3K inhibitors on NO production and the inflammatory response in the lung tissues of mice with viral pneumonia induced by IAV FM1 infection, C57BL/6 mice were divided randomly into control group, H1N1 group and LY294002 group. The models of viral pneumonia induced by infection with the IAV were established by nasal drops in the latter two groups. The LY294002 group was given the PI3K inhibitor LY294002 (i. p.) for 7 days from the day of modeling. Differences of hematoxylin and eosin (H&E) staining, H1N1-virus copy number, expression of the PI3K/AKT/eNOS signalling pathway, NO and inflammatory factors in lung tissues of the three groups of mice were compared. H&E staining showed obvious pathologic damage in the lung tissues of the H1N1 group, and the H1N1-virus copy number, expression of p-PI3K, pAKT, eNOS and contents of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α and intercellular adhesion molecule (ICAM)-1 were significantly higher than those of the control group ( P < 0.05). HE staining showed pathologic damage to be alleviated in the lung tissues of mice in the LY294002 group, and the expression of p-PI3K, p-AKT, eNOS and contents of IL-1β, IL-6, TNF-α, ICAM-1 were significantly lower than those of the H1N1 group ( P < 0.05). There was no significant difference in H1N1-virus copy number compared with that in the H1N1 group ( P>0.05). The present study suggests that a PI3K inhibitor can inhibit NO production and the inflammatory response in the lung tissues of mice with viral pneumonia induced by IAV FM1 infection.
16. Anti-inflammatory and synovial-opioid system effects of electroacupuncture intervention on chronic pain in arthritic rats
Chinese Acupuncture & Moxibustion,Part 3: Mechanism Study,Vol 35,No. 16
Objective To observe the analgesic effect of electroacupuncture (EA) on collagen-induced arthritis (CIA) rats and its regulating effect on inflammation reaction and the endogenous opioid system of synovial tissues. Methods A total of 30 healthy male Wistar rats were randomly divided into control group, model group and EA group, 10 rats in each one. The chronic pain model of CIA rats was made by bovine type-II collagen in the model group and EA group. Rats in the EA group were treated with EA at “Zusanli” (ST 36) and “Kunlun” (BL 60) for 30 min from 16 th day after model establishment, once a day for consecutive 10 days. Rats in control group did not receive any treatment. Rats in model group were treated with fixation as the EA group. Threshold of pain, arthritis index, paw swelling were measured before model establishment and 16 d, 20 d, 23 d and 25 d after model establishment. The levels of beta-endorphin (β-END), met-enkephalin (met-ENK), dynorphin A (Dyn A) were measured by radioimmunoassay; the mRNA expressions of mu opioid receptor (MOR), kappa opioid receptor (KOR) and delta opioid receptor (DOR) in synovial tissues of CIA rats were detected by quantitative polymerase chain reaction (qPCR). Results Compared with the control group, threshold of pain was reduced (all P < 0.01), arthritis index was increased (all P < 0.01) and paw swelling was increased (all P < 0.01) in the model group on the 16 th day, 20 th day, 23 rd day and 25 th day after model establishment. Compared with the model group, the threshold of pain was increased in the EA group (all P < 0.01), arthritis index and paw swelling were reduced (all P < 0.01) on the 23 rd day and 25 th day after model establishment. Compared with the control group, the level of Dyn A in synovial tissues of CIA rats was increased in the model group ( P < 0.01); the mRNA expressions of MOR, KOR and DOR were down-regulated lower than 0.5 fold of normal level. Compared with the model group, the level of β-END in synovial tissues of the knee joint was increased in the EA group ( P < 0.05), and the mRNA expressions of MOR, KOR and DOR in synovial tissues of CIA rats were up-regulated more than 2 folds of normal level. Conclusion The intervention of EA on chronic pain of CIA rats is superior, which is likely to be related with effects of EA on anti-inflammation and up-regulation of synovial tissue β-END and MOR, KOR, DOR.
17. Effects of Wine-processing on Rhei Radix et Rhizoma on Upper-energizer Disease and Effects on Activities of Energy Metabolism Enzymes in Liver
Journal of Chinese Medicinal Materials,Part 3: Mechanism Study,Vol 38,No. 17
Objective: To compare the effects of crude and wine-processed Rhei Radix et Rhizoma on upper-jiao disease and hepatic energy metabolism in mice. Methods: The streptococcal pneumonia rats model and acetic acid burning mouth ulcers rats model were prepared and randomly divided into three groups: model group, crude Rhei Radix et Rhizoma group and wine-processed Rhei Radix et Rhizoma group. The pathologic changes were observed after the rats had been given intragastric administration with water extracts of crude and wine-processed Rhei Radix et Rhizoma respectively. The normal ICR mice were randomly divided into three groups: control group, crude Rhei Radix et Rhizoma group and wine-processed Rhei Radix et Rhizoma group. The influence of water extracts of crude and wine-processed Rhei Radix et Rhizoma on the activities of Na +, K +-ATPase, Ca 2 +-ATPase and succinic dehydrogenase(SDH) in the mice were compared. Results: Compared with the crude one, the wine-processed Rhei Radix et Rhizoma significantly decreased the inflammation scores ( P < 0.05), and promoted the tissue repair of acetic acid burning mouth ulcers rats model. The wine-processed one could also obviously reduce and normalize the level of leucocyte and neutrophilic granulocyte; reduced the TNF-α level remarkably ( P < 0.05), and relieve inflammatory exudation of the alveolus pulmonis. The inhibitory effects of wine-processed Rhei Radix et Rhizoma on the activities of SDH, Ca 2 +-ATPase and Na +, K +-ATPase were weaker than those of the crude one ( P > 0.05). Conclusion: After wine-processing, the efficacy of Rhei Radix et Rhizoma on upper-jiao disease is enhanced, and the inhibition on the activity of energy metabolism enzyme in liver tends to be weakened.
18. Comparative Study on Effects of Anti-contusion Injury, Analgesia and Anti-inflammation of Root and Stem of Zanthoxylum nitidum
Journal of Chinese Medicinal Materials,Part 3: Mechanism Study,Vol 38,No. 18
Objective: To provide the scientific evidence for expansion of medicinal parts of Zanthoxylum nitidum by comparing the effects of anti-contusion injury, analgesia and anti-inflammation of its root and stem. Methods: The pharmacological effects between root and stem of Zanthoxylum nitidum were compared by observing the anti-injury effect in rats with injury struck by hammer. The analgesic effect in mice was evaluated by writhing test and hot plate test, and the anti-inflammatory effect on paw edema induced by carrageenan and granuloma induced by cotton pellet were investigated in rats. Results: Both root and stem of Zanthoxylum nitidum relieved the exterior and histological symptoms of rats' injury legs struck by hammer, decreased the numbers of mice's writhing, enhanced pain threshold of mice on heat plate, inhibited the edema of rats induced by carrageenan, and suppressed the granuloma of rats induced by cotton pellet. Conclusion: Stem of Zanthoxylum nitidum has similar effects of anti-contusion injury, analgesia and anti-inflammation with root of Zanthoxylum nitidum.
19. Study on the active fractions from Aegiceras corniculatum (L.) Blanco leaves for the anti-inflammatory and analgesic activities
Journal of Chinese Medicinal Materials,Part 3: Mechanism Study,Vol 38,No. 19
Abstract Objective: This study is aimed to screen and verify the effective fractions of Aegiceras corniculatum (L.) Blanco leaves for the anti-inflammatory and analgesic activities. Methods: Aegiceras corniculatum (L.) Blanco leaves were extracted by 75% ethanol to obtain five fractions, petroleum ether fraction, dichloromethane fraction, ethyl acetate fraction, n-butanol fraction and aqueous fraction. Acetic acid-induced writhing test in mice screened out ethyl acetate fraction with stronger anti-inflammatory and analgesic effects, which were then comprehensively evaluated through a series of tests in mice such as acetic acid-induced writhing test, the hot-plate test, carrageenan-induced paw edema and xylene-induced ear edema. Spectrophotometer was used to detect the total superoxide dismutase (T-SOD) activity and malondialdehyde (MDA) content in liver homogenate of mice with carrageenan-induced paw edema. Results: After i.g. administration of high dose (200 mg/kg) ethyl acetate fraction of Aegiceras corniculatum (L.) Blanco leaves, acetic acid-induced writhing times of mice were significantly reduced, pain threshold of mice in the hot-plate test increased, and the degrees of carrageenan-induced paw edema and xylene-induced ear edema were inhibited ( P < 0.01). Three days after i.g. administration of ethyl acetate fraction of Aegiceras corniculatum (L.) Blanco leaves of different doses (100, 150 and 200 mg/kg), T-SOD activity was significantly elevated and MDA content significantly decreased in the liver of mice with carrageenan-induced paw edema (both P < 0.01). Conclusion: Ethyl acetate fraction of Aegiceras corniculatum (L.) Blanco leaves has significant anti-inflammatory and analgesic effects and antioxidant effect may be one of the mechanisms underlying the anti-inflammatory effect.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 32,No. 20
Aim To investigate the anti-neuroinflammatory activities of dehydromiltirone and the underlying mechanisms in LPS-stimulated microglial cell line BV2 cells．Methods BV2 cells were pre-treated with dehydromiltirone, then stimulated by LPS. The levels of nitric oxide (NO) were measured by Griess assay, and the concentrations of pro-inflammatory cytokines were measured by ELISA assay. Confocal fluorescence microscopy was used to measure the expression of MAC-1, the biomarker of activated BV2 cells. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), NF-κB and PI3K/Akt were determined by Western blot analysis. Results The treatment of dehydromiltirone significantly inhibited the production of NO, TNF-α and IL-6, attenuated the expression of iNOS and COX-2 protein, and dampened the microglial activation in LPS-stimulated BV2 cells. The mechanistic study revealed that dehydromiltirone inhibited the phosphorylation of PI3K and Akt in LPS stimulated BV2 cells，and decreased NF-κB activation by suppressing the degradation of IκB. Conclusion Dehydromiltirone shows significant anti-neuroinflammatory effects through inhibiting PI3K/Akt phosphorylation and then inhibiting NF-κB signaling pathway．
21. Research on Effects of Ermiao Pill on Levels of Inflammation Mediators in Serum in Rats Gouty Model
Liaoning Journal of Traditional Chinese Medicine,Part 3: Mechanism Study,Vol 43,No. 21
Objective: On the basis of Ermiao Pill reducing the serum uric acid and presenting an anti-arthrolithiasis effect, to investigate the effects of Ermiao Pill on inflammation mediators in serum of rat gout model. Methods: Establish rat model of acute gouty arthritis with ehtambutol and adenine and carry out studies on pharmacology and efficacy of Ermiao Pill. Capillary electrolithiasis (CE) method was used to determine the content of serum protein. Results: Ermiao Pill can reduce the serum uric acid and presents an anti-arthrolithiasis effect and the results indicate that arthrolithiasis is related to the content of serum protein. Conclusion: From the effect of Ermiao Pill on inflammation mediators in serum in rat gout model, it can provide a scientific basis on the mechanism of Ermiao Pill anti-arthrolithasis.
22. Anti-inflammatory effect of Chikusetsu oleanane saponin on RAW264.7 cell through regulating SIRT1 activity
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 32,No. 22
Aim To investigate SIRT1 activity and anti-inflammatory effects of Chikusetsu oleanane saponin (COS) on the RAW264. 7 macrophage cells induced by lipopolysaccharide (LPS). Methods The nitric oxide (NO) release was detected with the Griess method. Western blot was used to detect the expression of tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β). Nuclear factor-κB (NF-κB) and SIRT1 were tested by immune- fluorescence. Results 1 mg/L LPS co-cultured with COS at 25-30 mg/L had no significant effect on the growth of RAW264. 7 cells. Compared with the LPS group, COS effectively inhibited the NO release and suppressed the expression of TNF-α and IL-1β; besides, it also inhibited the translocation of NF-κB and up-regulated the expression of SIRT1. Conclusion COS has protective effects on RAW264. 7 cells stimulated by LPS, which may be related to up-regulating the expression of SIRT1 and promoting the deacetylation of NF-κB, thereby inhibiting the translocation of NF-κB and reducing the expression of TNF-α and IL-1β.
23. Comparison on cellular anti-inflammatory and immunoregulatory activities of seven derivative combinations of Yaotongning with alkaloid as main part and in vivo pharmacodynamic evaluation on optimized candidate combination
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 47,No. 23
Objective To investigate which kind of Chinese medicinal effective fractions will produce good comprehensive cellular pharmacological activities when they were combined with alkaloids from Strychnos nux- vomica and Ephedra sinica, and to evaluate in vivo pharmacological activities of the effective fraction combination screened by cellular experiments. Methods The equal proportion mixture of alkaloids from Nux vomica and E. sinica was set as a sample (sample 1, A); Two samples (samples 2 and 3) were designed by respectively adding the equal ratio mixture of five flavonoids (F) at 50% into alkaloids from Nux vomica or E. sinica; Four samples (samples 4–7) were designed by respectively adding F, equal proportion mixture of four saponins (S), equal ratio mixture of six volatile oils/aqueous (V), and equal proportion mixture of six polysaccharides (P) at 50% into A. Murine macrophage cells and chondrocytes were exposed to the new recipes, and then the half inhibitory concentration (IC 50) of recipes for inhibiting (PGE 2) in macrophages and the half effective concentration (EC 50) for promotion of the secretion of IL-6, IL-1β, and TNF-α in macrophages were detected and compared. The interactions among the active fractions were evaluated by comparing the experimental EC 50/IC 50 values to their corresponding additive EC 50/IC 50 values calculated by the least square optimum method. The in vivo pharmacodynamics of the best combination was evaluated by the ear swelling model and delayed type hypersensitivity (DTH) model in mice. Results Sample 4 has good comprehensive activities of cellular anti-inflammation and immunoregulation. Moreover, strong synergistic effect among the effective fractions in this sample was observed; Cellular anti-inflammatory activities of samples 5, 6, and 7 were equivalent with sample 4; Samples 5 and 7 had good comprehensive cellular immunoregulation activity; But the alkaloids mixture (A) and the combinations of Nux vomica or E. sinica alkaloids with F (samples 2 and 3) were significantly weaker than sample 4 in cellular anti-inflammation, immunoregulation, and chondrocyte-proliferation. Sample 4 also exhibits a certain effect on in vivo anti-inflammation and immunity in mice when A was decreased at 25% or 5%. Conclusion It is not suitable to design a combination just by alkaloids from Nux vomica and alkaloids of E. sinica; Alkaloids from Nux vomica or alkaloids of E. sinica are also not appropriate to solely combine with the mixture of flavonoids (F). When A is combined with F, S, V, and P, respectively, synergistic or additive effects among the active fractions are usually observed. These active fractions help to strengthen comprehensive cellular pharmacological activities of A. Sample 4 not only has good cellular activities of anti-inflammation and immunoregulation but also has better in vivo effect on anti-inflammation and immunity, suggesting that it is feasible to screen the optimized Chinese medicine formula based on cellular pharmacological experiments.
24. The Effects of Biyuanshu Oral Solution on the Molecular Chaperone HSP70 and the Cofactor CHIP for the Intervention of the Inflammation of Sinus Mucus Membrane in Mice CRS Model
Journal of Chinese Medicinal Materials,Part 3: Mechanism Study,Vol 39,No. 24
Objective: To investigate the effects of Biyuanshu (BYS) on molecular chaperone HSP70 and carboxyl terminus of HSC70/HSP70-interacting protein (CHIP) expression of nasal sinuses mucosa epithele in mice Chronic rhinosinusitis (CRS) model, and to explore the BYS intervention mechanism from the point of molecular chaperone system. Methods: 140 C57 male mice were randomly divided into normal group, sham operation group, model group, western medicine group, BYS low-dosage group, BYS medium-dosage group, BYS high-dosage group, with 20 mice in each group, and CRS model was established. With corresponding drug treatment for 14 days. Nasal sinuses mucosa tissue was collected to observe pathological alterations after HE dyeing, and HSP70 and its cofactor CHIP mRNA expression in nasal sinuses mucosa epithele were detected by real-time PCR, and the protein expression and IKK activity were detected by Western blotting. Results: Model group appeared large necrotic and falling-off areas, apparently accompanied with chronic inflammatory cell infiltration. Nasal sinuses mucosa epithelial chaperon HSP70 and its cofactor CHIP expression levels were much lower in CRS group than normal group and slam operation group ( P < 0.05 or P < 0.01), the p-IKK α/ β expression in model group was obviously higher than normal group and slam operation group ( P < 0.01). Compared to model group, the BYS medium-dosage and high-dosage groups presented well-repaired epithele in alignment, with fewer chronic inflammatory cell infiltration. Furthermore, the expression of chaperon HSP70 and its cofactor CHIP in nasal sinuses mucosa epithelium were much higher than model group ( P < 0.01), but the p-IKK α/β expression was lower ( P < 0.01). Conclusion: BYS can upregulate chaperon HSP70 and its cofactor CHIP to enhance intracellular protection from inflammatory protein injury mice, and reduce IKK activity to intervene on downstream NF-κB signaling pathway. BYS can be in favor of nasal sinuses mucosa epithelial repairmen.
25. Substitutability of Prunella vulgaris L. Stem Leaf and Ear Based on HPLC-ESI-MS n Analysis and Anti-Inflammatory and Antioxidant Activity
Chinese Pharmaceutical Journal,Part 3: Mechanism Study,Vol 51,No. 25
OBJECTIVE To compare the chemical composition and therapeutic effect between Prunella vulgaris L. stem leaf and ear, thus to provide evidence for judging whether the stem leaf of Prunella vulgaris L. can substitute the ear as an herbal medicine. METHODS Aqueous extracts of the stem leaf and ear of Prunella vulgaris L. from different producing areas were analyzed with HPLC-ESI-MS n. The anti-inflammatory effects were observed by inflammatory models of ear edema induced by dimethylbenzene in mice and hind paw edema induced by carrageenan in rats. Enzyme-linked immunosorbent assay (ELISA) was used to detect the plasma level of TNF-α and antioxidant activities were detected by ABTS method. RESULTS Prunella vulgaris L. stem leaf and ear were not significantly different in chemical composition, both of which contained mainly triterpenoids, flavonoids, phenolic acids and other substances. Compared with the model group, Prunella vulgaris L. ear significantly reduced the hind paw edema in rats induced by carrageenan from 1 h after oral administration ( P < 0.05), while the onset time of stem leaf was later than 1 h. Both groups could significantly reduce the ear edema in mice induced by dimethylbenzene ( P < 0.01). The TNF-α levels in the Prunella vulgaris L. stem leaf and ear groups [(24.16 ± 1.24) and (24.33 ± 2.36) ng·mL ?1] were lower than that in the model group [(31.34 ± 1.94) ng·mL ?1] ( P < 0.01). Prunella vulgaris L. stem leaf and ear groups showed strong antioxidant activities in the ABTS · + scavenging test. CONCLUSION The content of the main constituents in Prunella vulgaris L. stem leaf and ear has significance differences. The results of animal tests indicate that the aqueous extracts of Prunella vulgaris L. stem leaf and ear have significant anti-inflammatory and antioxidant effects.
Acta Pharmaceutica Sinica,Part 3: Mechanism Study,Vol 51,No. 26
To study the chemical composition and their anti-inflammatory activities of honeysuckle (Lonicera japonica Thunb.) roots, seventeen compounds were isolated from the roots of L. japonica Thunb. by various chromatography, including silica gel, Sephadex LH-20 and preparative HPLC. Their structures were identified by MS, IR, and nuclear magnetic resonance spectra, as 1-oxo-(1H)-cyclopenta [b] benzofuran-7-carbaldehyde (1), 4-hydroxycinnamic acid (2), chlorogenic acid (3), loganin aglycone (4), caffeic acid (5), secologanin dimethyl acetal (6), korolkoside (7), coniferin (8), sweroside (9), secoxyloganin (10), 5-O-caffeoylquinic acid (11), chlorogenic acid methyl ester (12), chlorogenic acid ethyl ester (13), 3,5-O-dicaffeoylquinic acid (14), 4,5-O-dicaffeoylquinic acid (15), grandifloroside (16), and 4,5-O-dicaffeoylquinic acid (17). Among those, compound 1 is a new compound, and compound 8 is found in L. japonica for the first time. Compounds 1, 3, 14–17 showed significant anti-inflammatory activities against macrophage in zebrafish.
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 47,No. 27
Objective To research the chemical composition from the roots of Lonicera japonica and their anti-inflammatory activities. Methods The compounds were isolated and purified by chromatography on silica gel column, Sephadex LH-20 column preparative thin-layer, and semi-preparative HPLC, etc. Their structures were identified on the basis of spectroscopic data, and part of the isolated compounds were tested for their anti-inflammatory activities against zebrafish. Results Fourteen compounds were isolated and elucidated as 3,13-dihydroxystemodan-2-one (1), chrysophanol (2), palmarumycin CP 2(3), β-sitosterol (4), stigmastero (5), stigmast-4,6,8 (14), 22-tetraen-3-one (6), erythrinassinate D (7), lanosterin (8), 3-O-acetyloleanolic acid (9), protocatechuin aldehyde (10), daucosterol (11), (E)-3-(3,4-dihydroxybenzylidene)-5-(3,4-dihydroxyphenyl)-2(3H)-furanone (12), lomacarinoside B (13), and (2E,6S)-8-(α- L-arabinopyranosyl-(1″→6′)-β- D-glucopyranosyloxy)-2,6-dimethyloct-2-eno-1,2″-lactone (14). Conclusion Compound 1 is a new compound,and compounds 2,3,6,7–9, and 13 are isolated from L. japonica for the first time. Compounds 9 and 10 show the significant anti-inflammatory activities at 100 μg/mL.
28. Effects of Hedyotis diffusa Extract on Epidermal Growth Factor Induced Proliferation, Apoptosis, and TNF-α Induced Inflammatory Factors of HaCaT Cells
Chinese Journal of Integrated Traditional and Western Medicine,Part 3: Mechanism Study,Vol 36,No. 28
Objective To observe the effects of Hedyotis diffusa extract (HDE) on the proliferation, apoptosis, and inflammatory factors of HaCaT cells, and to explore its possible molecular mechanisms. Methods HaCaT cells were divided into 3 groups, the blank group, the control group, and 3 dose HDE groups. No epidermal growth factor (EGF) or HDE was added in the blank group. EGF was added with no HDE treatment in the control group. HDE (25, 50, 100 mg/mL) and EGF were added in the 3 HDE groups to co-culture HaCaT cells. The effects of HDE on EGF-induced proliferation of HaCaT cells were detected using CCK-8 assay. The effects of HDE on the growth cycle and apoptosis rate of HaCaT cells were measured using flow cytometry. Moreover, the protein expression levels of Ki67, Bcl-XL, cIAP1, procaspase-3, and cleaved caspase-3 were determined using Western blot. In addition, the levels of IL-6, IL-8, and IL-10 in the supernatant were detected using ELISA. The level of phosphorylation of NF-κB p65 (p-p65) was measured using Western blot. Results Compared with the blank group, the absorbance of HaCaT cells and the expression level of Ki67 increased ( P < 0.05, P < 0.01); the levels of p-p65, IL-6, and IL-8 were elevated ( P < 0.05, P < 0.01); the IL-10 level was lowered ( P < 0.01) in the control group. Compared with the control group, the absorbance of HaCaT cells and the expression level of Ki67 decreased ( P < 0.05, P < 0.01); the levels of p-p65, IL-6, and IL-8 were reduced ( P < 0.05, P < 0.01); the IL-10 level was elevated ( P < 0.05, P < 0.01) in the 3 dose HDE groups. Meanwhile, the apoptosis rate of HaCaT cells increased more in the 3 dose HDE groups than in the control group (P < 0.05, P < 0.01). The percentage of HaCaT cells at G1 phase was 58.51%, 73.12%, and 79.95% in 25, 50, and 100 mg/mL HDE groups, respectively, showing statistical difference when compared with that in the control group ( P < 0.05, P < 0.01). The percentages and apoptosis rates of HaCaT cells at G1 phase were elevated more in 50 and 100 mg/mL HDE groups than in 25 mg/mL HDE group ( P < 0.01). Besides, the percentage and apoptosis rate of HaCaT cells at G1 phase were elevated more in 100 mg/mL HDE group than in 50 mg/mL HDE group ( P < 0.05). Compared with the control group, the protein expression levels of Bcl-XL and cIAP1 were reduced in 100 mg/mL HDE group ( P < 0.01). There was no statistical difference in caspase-3 total amount ( P > 0.05), but the cleaved caspase-3 expression increased with statistical difference ( P < 0.01). Conclusion HDE inhibited the proliferation of HaCaT cells possibly by arresting HaCaT cell growth at G1 phase, promoted the apoptosis of HaCaT cells by stressing protein expressions of Bcl-XL and cIAP1, and elevating protein expressions of cleaved caspase-3, and suppressed inflammatory response of HaCaT cells via regulating NF-κB expression.
29. Curcumin analog exhibited anti-inflammatory activity through inhibiting ERK/JNK and NF-κB signaling pathway
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 47,No. 29
Objective To study the anti-inflammatory activity and mechanism of mono-carbonyl curcumin analog 22. Methods The cell cytotoxicity of compound 22 was detected by MTT assay. LPS activated peritoneal macrophages was used as cell model. The effect of compound 22 on inflammatory cytokines protein and gene expression was detected by ELISA and RT-q PCR, respectively. UV-absorbing assay was used to determine the stability of compound 22. The anti-inflammatory mechanism of compound 22 was detected by Western blotting assay. Results Compound 22 showed no cytotoxicity after incubated cells 24 h. Compared with curcumin, compound 22 dose-dependently inhibited LPS-induced production of inflammatory cytokines TNF-α and IL-6. Compound 22 showed more stability than curcumin in vitro. Meanwhile, compound 22 inhibited LPS-induced inflammatory gene IL-1β, IL-12, IL-6, and TNF-α expression through q RT-PCR assay. Compound 22 inhibited the phosphorylation of ERK/JNK, also reversed the LPS-induced degradation of IκB. Conclusion Compound 22 exhibits its anti-inflammatory activity through inhibiting ERK/JNK signaling pathway, as well as NF-κB activation.
30. Protective effects of salvianolic acid B on isoproterenol induced myocardial ischemic rats and its relation with NLRP3 expression
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 32,No. 30
Aim To evaluate the protective effects of salvianolic acid B on the ISO-induced myocardial ischemic injury model of rats and the influence of regulating NLRP3 associated protein on myocardial ischemia. Method All rats were randomly divided into control group, model group and Sal B 5, 10, 15 mg·kg ?1 groups. For 7 days, rats in Sal B groups were given by introperitoneal injection of 5, 10, 15 mg·kg ?1 Sal B, rats in control group and model group were given the same volume of normal saline. Rats were subcutaneously multi-point injected ISO (30 mg·kg ?1) for 2 days on the fifth administrating day. The myocardial protective effect of Sal B was evaluated from electrocardiogram (ECG), myocardial tissue pathological changes, serum myocardial enzymes, oxidation index and inflammatory cytokine, myocardial tissue of NLRP3related protein expression. Results Sal B could reduce the degree of myocardial tissue necrosis and the infiltration of inflammatory cells, reduce T-wave values of ECG ( P < 0.05 or P < 0.01). Compared with model group, CK values, GOT values and IL-1β values of rats in different dose groups were significantly lower, and MDA values and LDH values of rats in middle-and high-dose groups were significantly lower ( P < 0.05 or P < 0.01). However, T-SOD values of rats middleand high-dose groups were significantly higher ( P < 0.05 or P < 0.01). Meanwhile, the NLRP3, Caspase-1 and IL-1β protein level in myocardial tissue of the rats in different dose groups compared with model group had reduced ( P < 0.05 or P < 0.01). Conclusion Sal B has protective effects on myocardial ischemic rats, its mechanism may be related with inhibition of decreasing the expression of NLRP3 inflammasome associated protein, which can suppress the generation of inflammatory cytokines.
31. Rhubarb Peony Decoction Ameliorates DSS-induced Ulcerative Colitis and It’s Associated Anemia in Mice
Chinese Archives of Traditional Chinese Medicine,Part 3: Mechanism Study,Vol 34,No. 31
Objective: To study the therapeutic effect of Rhubarb Peony Decoction (RPD) on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) and UC-associated anemia in mice. Methods: The normal control group, UC group, mesalazine group, low dose group of Rhubarb Peony Decoction (RPDL), middle dose group of Rhubarb Peony Decoction (RPDH), and high dose group of Dahuang Mudan Decoction (DMDH) were designed. The disease activity index (DAI), the length of the colon, histological analysis, colon pathological lesion scores, colon myeloperoxidase (MPO) activity as well as the number of peripheral blood neutrophils, the total number of red blood cells, and hemoglobin content were investigated to evaluate the treatment effect. Results: Compared with the normal control group, DAI scores of UC group were significantly increased. The cross-section of the colon was significantly thicker and accompanied with a large number of inflammatory cell infiltration, multifocal ulceration, crypt and intestinal glands reduced, histological scores and colon MPO activity significantly high; neutrophil cells in peripheral blood increased significantly; the number of red blood cells and hemoglobin was all significantly reduced. Compared with the DSS group, mesalazine reduced DAI scores and histological scores, but the effect on colon thickening, neutrophils increasing, red blood cells and hemoglobin reduction in UC mice was not obvious. All doses groups of RPD can lower DAI scores, histological scores, and colon MPO activity. RPDM and RPDH also can significantly prevent colon tissue thickening, maintain the integrity of its structure, inhibit neutrophils increase in peripheral blood, increase red blood cells and hemoglobin content significantly. Conclusion: Rhubarb Peony Decoction has therapeutic effect on DSS-induced UC and UC-associated anemia in mice.
32. Spectrum-Activity Relationships between HPLC Fingerprints and Analgesic and Anti-inflammatory Activities of Different Extracts of Asari Radix Et Rhizoma
Liaoning Journal of Traditional Chinese Medicine,Part 3: Mechanism Study,Vol 43,No. 32
Objective: To study the relationships between HPLC fingerprints and analgesic and anti-inflammatory activities of different extracts of Asari Radix Et Rhizoma. Methods: HPLC fingerprints of different extracts of Asari Radix Et Rhizoma were established. The hot plate-induced pain and xylene-induced ear swelling experiments were carried out to test the analgesic and anti-inflammatory activities of different extracts of Asarum. Partial least-squares regression analysis was taken to analyze the correlation of these spectrum peaks and its analgesic and anti-inflammatory activities in different extracts. Results: There were 15 peaks positively correlated with analgesic activity among 31 characteristic HPLC peaks. Peak No. 8, 11, 13, 20, 21, 22 and 26 played more important parts. And there were 20 peaks positively correlated with anti-inflammatory activity. Peak No. 2, 5, 7, 9, 10 and 11 played more important parts. Conclusion: There were characteristic effect ingredients in Asari Radix Et Rhizoma for analgesic and anti inflammation activities.
33. Total flavonoids of Epimedium attenuate aging-related inflammation in rat brain by inhibiting MAPK/NF-κB signaling pathway
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 33,No. 33
Aim To investigate the effect of the total flavonoids of Epimedium (TFE) on MAPK/NF-κB signaling pathway and the inflammatory reaction in the hippocampus of natural aging male rats. Methods The morphological changes of the hippocampus composed of three areas (CA1, CA3 and DG) were observed using haematoxylin-eoin (HE) staining. The protein expression levels of senescence-associated protein p21, apoptosis-related proteins Bax and Bcl-2, nuclear transcription factor-κB p65 (NF-κB p65) and its downstream inflammatory factors TNF-α, IL-1β and COX-2, and MAPK signaling pathway-related proteins (ERK1/2, p-ERK1/2, JNK, p-JNK, p38 MAPK, p-p38 MAPK) in hippocampal were detected by Western blot. Results Compared with natural aging group, TFE obviously improved the morphology and structure of hippocampal neurons, and the nerve cells arranged neatly and closely. Furthermore, TFE significantly downregulated the protein expression levels of p21 and Bax, upregulated the protein expression levels of Bcl-2 and the ratio of Bcl-2/Bax, and reduced the expression of NF-κB p65 and of its downstream inflammatory factors TNF-α, IL-1β, COX-2, and MAPK signaling pathway-related proteins (p-ERK1/2, p-JNK and p-p38 MAPK) in hippocampus of natural aging rats. Conclusions TFE effectively protects against inflammatory reaction in brain aging of SD male rats. The mechanism is related with inhibition of NF-κB nuclear translocation and reduction of its downstream inflammatory cytokines expression by inhibiting MAPK signaling pathway activation.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 33,No. 34
Aim To study the effects of menthone on LPS-induced inflammation in mice and to explore the mechanisms of activating of NLRP3 inflammasome. Methods C57BL/6J male mice were randomly divided into five groups, namely the normal group, model group, dexamethasone group (5 mg·kg ?1) and menthone (0.25, 0.5 g·kg ?1) groups. The dexamethasone group was given the drugs once by intraperitoneal injection on 5th day, while the other mice were given drugs by oral administration once a day for 5 d. Then, the normal group was injected with the saline and the other groups were injected LPS (15 mg·kg ?1, 0.01mL·g ?1) after 30 min of the last administration. 12 h after LPS injection, the blood, serum, and lung tissue of mice were collected. The levels of IL-18, IL-1β, IL-5, TNF-α, IFN-γ, G-CSF, GM-CSF and MIP-1β were measured in serum by ELISA and Luminex Magpix. The white cells in blood (WBC) were counted. The lung index and the levels of NO in lung tissue were calculated. The lung histopathology was performed. The relative levels of NLRP3, caspase-1 and IFN-αmRNA in lung tissue were determined by RT-PCR. The expressions of cathepsin B and P2X7R in lung tissue were quantified by immunohistochemical analysis. Results 0.5 g·kg ?1 menthone significantly reduced the levels of IL-18, IL-1β, IL-5, TNF-α, IFN-γ, G-CSF, GM-CSF, and MIP-1β in serum ( P < 0.05 or P < 0.01), and the expression of NLRP3, IFN-α, cathepsin B, P2X7R, NO in lung tissue ( P < 0.05 or P < 0.01). 0.25 g·kg ?1 menthone inhibited the IL-1β, IL-5, TNF-α, MIP-1β production in serum and the expression of P2X7R, NO in lung tissue ( P < 0.05). 0.25, 0.5 g·kg ?1 menthone could attenuate pathological change of the lung tissues by reducing neutrophil infiltration and thickening of alveolar septa. And it slightly reduced the lung index and the level of white cells in blood. Conclusion Menthone has a protective effect on LPS-induced inflammation mice, and its mechanism is related to inhibiting the release of varies of inflammatory cytokines, reducing the inflammation reaction and inhibiting the activation of NLRP3 inflammasome.
35. Regulatory Effects of Total Saponins from Panax japonicus on Colonic Inflammatory Response Through the Endoplasmic Reticulum Stress Responses in Aging Rats
Journal of Chinese Medicinal Materials,Part 3: Mechanism Study,Vol 40,No. 35
Objective: To investigate the effects of total saponins of Panax japonicus (SPJ) on inflammatory response in the colon of aging rats through reticulum endoplasmic stress reaction. Methods: Male SD rats were divided into adolescent group (6 months), aging group (24 months), SPJ low-, medium-and high-dose groups, and rats of 24 month were intervened by the SPJ in different dose groups. The serum level of D-lactic acid in rats were detected by D-lactic acid assay kit. The mRNA expression levels of inflammatory factors IL-1 β and TNF- α were detected by RT-PCR. The expression of COX-2 and endoplasmic reticulum stress response marker protein GRP78, CHOP and p-eIF2 α were detected by Western Blot. Results: Compared with the young group, the serum level of D-lactic acid, and the mRNA expression levels of the inflammatory factors IL-1 β and TNF- α, the expression of COX-2, and endoplasmic reticulum stress response marker protein GRP78, CHOP and p-eIF2 α were significantly increased in the aging group. However, compared with the aging group, the serum level of D-lactic acid, the mRNA levels of inflammation factors IL-1 β and TNF- α, and the expression of COX-2 and endoplasmic reticulum stress response marker protein GRP78, CHOP and p-eIF2 α in SPJ pretreatment groups were significantly decreased. Conclusions: Intestinal mucosal barrier dysfunction exist in aging rats, inflammation and endoplasmic reticulum stress response exist in the colon of aging rats. Total saponins of Panax japonicus can reduce intestinal inflammation, and the mechanism may be related to the reduction of endoplasmic reticulum stress responses.
36. Protective effects of salvianolic acid on myocardial ischemic injury of rats from the aspect of inhibiting inflammatory reaction
The Chinese Journal of Clinical Pharmacology,Part 3: Mechanism Study,Vol 33,No. 36
Objective To investigate the protective effects of salvianolic acid on myocardial ischemic injury of rats from the aspect of inhibiting inflammatory reaction. Methods Sixty Sprague Dawley (SD) male rats were randomly divided into four groups ( n = 15 per group): sham operation group, model group, experimental-high (36 mg·kg ?1) group, experimental-low (18 mg·kg ?1) group with intraperitoneal injection dosing. Same amount of distilled water was injected to sham operation group and model group, lasting for 4 days. Myocardial ischemia model was established by coronary artery ligation of left anterior descending (LAD) branch after the last administration. Six hours after the operation, the blood of eye canthus was collected to assay the levels of creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB) and lactate dehydrogenase (LDH) in serum. The heart was collected 24 h after the operation, stained by hematoxylin eosin (HE) and checked the pathological change of the cardiac muscle tissue, assayed the cardiac troponin (cTnT), tumor necrosis factor α (TNF-α) in serum and myeloperoxidase (MPO) was measured in myocardial tissue homogenate. Results After 6 hours’ myocardial ischemia, the serum CK in sham operation group was (529.61 ± 141.93) U·L ?1, CK-MB was (708.12 ± 385.93) U·L ?1, LDH was (330.12 ± 158.38) U·L ?1and cTnT was (294.12 ± 55.10) pg·mL ?1 after 24 hours’ myocardial ischemia. The serum CK in model group was (996.42 ± 413.42) U·L ?1, CK-MB was (1346.11 ± 558.30) U·L ?1, LDH was (520.12 ± 154.76) U·L ?1 and cTnT was (513.00 ± 69.71) pg·mL ?1 after 24 hours’ myocardial ischemia. The serum CK in experimental-low and experimental-high groups were (499.45 ± 159.33), (514.91 ± 98.82) U·L ?1, CK-MB were (831.42 ± 385.11), (592.10 ± 206.32) U·L ?1, LDH were (462.62 ± 229.68), (437.72 ± 175.80) U·L ?1, and cTnT were (431.12 ± 106.00), (338.80 ± 76.92) pg·mL ?1 after 24 hours’ myocardial ischemia. Compared with the sham operation group, the CK, CK-MB and LDH value in the serum of model group increased significantly ( P < 0.05). Compared with the model group, the salvianolic acid significantly both can decrease the CK and CK-MB in serum (P < 0.05). Compared with sham operation group, the cTnT value in the serum of model group increased significantly ( P < 0.05). Compared with model group, high dosage of salvianolic acid could significantly decrease the cTnT value ( P < 0.05). After 24 hours’ myocardial ischemia, the TNF-α in serum of sham operation group was (118.90 ± 17.58) pg·mL ?1, the MPO in heart tissue was (32.25 ± 3.75) U·L ?1. The TNF-αin serum of model group was (156.00 ± 28.24) pg·mL ?1, the MPO in heart tissue was (104.83 ± 22.87) U·L ?1. The TNF-α in serum of experimental-low and experimental-high groups were (136.05 ± 34.00), (125.06 ± 30.13) pg·mL ?1, the MPO in heart tissue were (91.70 ± 21.57), (68.00 ± 18.47) U·L ?1. Compared with sham operation group, the TNF-αvalues in the serum and the MPO value in model group increased significantly ( P < 0.05). Compared with model group, these values of experimental-high group decreased with significantly (all P < 0.05). Conclusion Salvianolic acid showed the protection of myocardial tissue, which also could significantly reduce the myocardial cell injury and inflammatory cell infiltration of cardiac myocytes, which could be related with the decreased levels of TNF-α and MPO.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 33,No. 37
Aim To study the anti-inflammatory activity of the Channa argus bile. Methods The bile was isolated and purified by extraction and silica gel column chromatography. Then the compounds were identified by hydrogen and carbon spectra. The spleen lymphocytes proliferation assay and Lipopolysaccharide (LPS) induced mouse macrophage RAW264.7 releasing Nitrogen Monoxide (NO) experiment were used to evaluate the anti-inflammatory activity. Results Compound (C1) of sodium taurocholate and compound (C2) of sodium taurochenodeoxycholate were isolated by activity tracing. The cell relative viabilities of the two compounds on Concanavalin A (ConA) induced spleen lymphocytes proliferation assay were 65.9% ± 11.7% and 60.5% ± 9.4%, which were significantly different from the result of the model group ( P < 0.01), respectively. The NO production of LPS-induced RAW 264.7 release of NO was (16.4 ± 1.9) μmol·L –1 and (15.5 ± 1.7) μmol·L –1, which were significantly different from the result of model group ( P < 0.01). Conclusion Sodium taurocholate and sodium taurochenodeoxycholate from Channa argus perform the anti-inflammatory activities but have no cytotoxic effect on spleen lymphocytes and macrophage.
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 48,No. 38
Objective: To investigate the anti-inflammatory activity of Compound Mango Leaves Formula (CMLF) and preliminarily elucidate the network regulatory mechanism. Methods: UPLC-Q/TOF-MS combined with NF-κB dual-luciferase reporter assay systems was used to screen the anti-inflammatory compounds in CMLF. Network pharmacology approach was utilized to predict the targets and pathways. The anti-inflammatory effects were then verified by TNF-α-induced BEAS-2B cell inflammatory model and ELISA. Results: Sixteen anti-inflammatory compounds were screened from CMLF. The compounds acted on 40 target molecules and were related to 11 inflammation singaling pathways, respectively. Conclusion: The anti-inflammatory multi-layer regulation mechanism of CMLF is related to MAPK, focal adhesion and VEGF signaling pathways.
39. Variation analysis of flavonoids contents and nitric oxide production inhibition in different parts from Hypericum perforatum
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 48,No. 39
Objective To provide a chemometric analytical approach for classification of different parts from Hypericum perforatum using ultra performance liquid chromatography (UPLC) combined with chemometrics methods. Methods The ACQUITY UPLC ?BEH C 18 (50 mm × 2.1 mm, 1.7 μm) column was used, and 0.2% formic acid aqueous solution-acetonitrile was taken as gradient elution system. The chromatogram information of different parts including flower, fruit, leaf, stem and root from H. perforatum L. was collected. The original data were pretreated by centralization and normalization. With partial least squares discriminant analysis (PLS-DA) and PLS-tree cluster analysis, the half inhibition concentration of nitric oxide (NO) production activity was taken as an anti-inflammatory factor to evaluate the similarities and differences in flavonoid contents and nitric oxide production inhibition in different parts from H. perforatum. Results All the calibration curves of 6 flavonoids showed good linearity in each range with correlation coefficients greater than 0.999, indicating good precision, repeatability and stability, and the average recovery ranged from 97.28% to 102.84%. By PLS-DA and PLS-tree, according to the data of flavonoid contents and NO product inhibition activities, the quality was in the sequence of leaf > flower > stem > fruit > root of H. perforatum. Conclusion The established method suggested that an appropriate harvest part of H. perforatum is the stem part on the ground.
40. Effects of tripchlorolide on the expression of inflammatory factors and β-amyloid protein in the hippocampus in vascular dementia rats
The Chinese Journal of Clinical Pharmacology,Part 3: Mechanism Study,Vol 33,No. 40
Objective To investigate the effects of tripchlorolide on the accumulation of nuclear factor-κb (NF-κB), cyclooxygenase-2 (COX-2), β-amyloid protein (Aβ) in the hippocampus in vascular dementia (VD) rats. Methods Male Sprague-Dawley (SD) rats were randomly assigned into three groups (10 rats per group): sham operation group, model group and experimental group. The bilateralcornmoil carotid arteries ligation (2-VO) models were created in model and experimental rats. Tripchlorolide were dissolved in sterile 1% dimathyl sulfoxide (DMSO) and were administered (1 μg·kg -1·d -1) by intraperitoneal injection daily for 4 weeks in experimental group. In sham operation and model group rats were given DMSO in the same volume. The expressions of Aβ, NF-κB, COX-2 in the brain were detected by Western blot. Results Aβ protein bands of gray value in rat brain campus in sham operation group, model group and experimental group were 0.17 ± 0.05, 1.46 ± 0.06, 0.57 ± 0.04. NF-κB p65 protein bands of gray value in rat brain campus in sham operation group, model group and experimental group were 0.12 ± 0.03, 0.86 ± 0.04, 0.37 ± 0.04. COX-2 protein bands of gray value in rat brain campus in sham operation group, model group and experimental group were 0.23 ± 0.04, 1.26 ± 0.05, 0.73 ± 0.04. Compared with sham operation group, the three indexes in model group were increased significantly (all P < 0.001). Compared with model group, the three indexes in experimental group were decreased significantly (all P < 0.001). Conclusion Tripchlorolide exhibits anti-inflammatory and attenuates brain Aβ deposition in 2-VO rats.
41. Influences of scutellarin on ATP-induced inflammasome activation and pyroptosis in J774A.1 macrophages
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 34,No. 41
Aim To explore the influences of scutellarin on ATP-induced NLRP3 inflammasome activation and pyroptosis, using LPS-primed murine macrophages J774A.1 as an inflammatory cell model, and to explore the underlying mechanism. Methods The effects of scutellarin on ATP-induced pyroptosis in murine J774A.1 macrophages were analyzed by propidium iodide (PI) staining assay. The levels of IL-1β, caspase-1 and HMGB1 in cell lysates and culture supernatants were analysed using Western blot. The levels of IL-1β in cell culture supernatants were measured by cytometric beads array (CBA). Results ATP significantly induced caspase-1 activation and mature IL-1β and HMGB1 release into the culture supernatants in LPS-primed murine J774A.1 macrophages, and induced pyroptosis. Scutellarin treatment dose-dependently inhibited ATP-induced caspase-1 activation, mature IL-1β and HMGB1 release, and pyroptosis. Notably, scutellarin’s inhibitory effects on ATP-induced pyroptosis were markedly reversed by the adenylate cyclase inhibitor MDL12330A and selective protein kinase A (PKA) inhibitor H89. Conclusion Scutellarin inhibits NLRP3 inflammasome activation and pyroptosis by modulating the PKA activity in macrophages, thereby exhibiting anti-inflammatory activities.
42. Catechin remits the inflammation in allergic asthma mice by suppressing NF-κB-TSLP signal pathway
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 34,No. 42
Aim To explore the mechanism of catechin in inhibiting Th2-driven inflammation responses of allergic asthma mice. Methods Female BALB/c mice were chosen to establish an allergic asthma model by being sensitized and excited with OVA. Then, the mice in the administered group were intervened with catechin at different doses (75, 150, and 300 mg·kg ?1). The proportion of peripheral leukocyte cells was analysed, and pathological changes in lung tissues were observed by HE dyeing. The levels of TSLP, IL-5, and IL-13 in lung tissues and OVA-specific Ig E in mouse serum were measured using ELISA kit. NF-κB p65, and IκB protein expression levels in lung tissues were investigated using Western blot, and NF-κB p65 nuclear translocation was detected with immunofluorescent technique. Results Catechin could not only effectively reduce the proportion of peripheral leukocyte cells, but relieve the levels of TSLP, Th2 inflammatory factor IL-5 and IL-13, and the activation of NF-κB signal pathway in lung tissues of allergic asthma mice. Conclusion Catechin can effectively relieve Th2 inflammation in allergic asthma mice induced by allergen OVA. The possible mechanism may be that it could reduce expression of TSLP by inhibiting the NF-κB signal.
43. Effects of emodin on oxidative stress and inflammatory response in rats with acute spinal cord injury
China Journal of Chinese Materia Medica,Part 3: Mechanism Study,Vol 43,No. 43
This paper was aimed to investigate the effects of emodin on oxidative stress and inflammatory response in rats with acute spinal cord injury (SCI), and to explore the protective mechanism of emodin on neurons after SCI. Rat SCI models were established using a modified Allen’s method. One hundred and ninety five Sprague-Dawley (SD) rats were randomly divided into sham operation group (group A), model group (group B), 20 mg·kg ?1 emodin treatment group (group C), and 40 mg·kg ?1 emodin treatment group (group D), and 80 mg·kg ?1 emodin treatment group (group E). Functional recovery was evaluated using the basso-beattie-bresnahan (BBB) scale and inclined plate test on the 3rd, 7th, 14th and 28th days. On the 7th day after SCI, Nissl bodies in the neurons were observed after toluidine blue staining. The changes of myelinated nerve fibers were observed by transmission electron microscopy. Nrf2, HO-1, NQO1, GFAP, NF-κB protein expression levels were detected by Western blot. The contents of TNF- α, IL-1 β, and IL-6 were detected by ELISA. Immunofluorescence staining was performed to observe the expression of the GFAP, NG2 and ED-1. The BBB and inclined plate scores of group C, D and E were higher than those in group B on the 7th, 14th, and 28th days, and the difference was statistically significant ( P < 0.05). On the 7th day, Nissl bodies of groups B and C started to fuse, and the fusion of groups D and E was significantly alleviated than that in groups B and C. Transmission electron microscope showed that the changes of demyelination were obvious in groups B and C, those in groups D and E were significantly improved as compared with those in group B and C. Western blot showed that the differences in Nrf2, HO-1, GFAP, and NF-κB protein expression between groups C, D, E and group B were statistically significant, and the differences in NQO1 protein expression between groups D, E and group B were also statistically significant ( P < 0.05). ELISA showed that the differences in the contents of TNF- α, IL-6 between groups C, D, E and group B were statistically significant, and the differences in IL-1 β contents between groups D, E and group B were also statistically significant ( P < 0.05); Immunofluorescence assay showed that the expression levels of GFAP and NG2 in groups C, D and E were higher than those in group B, and more obvious elevations were observed in groups D and E. The expression of ED-1 in groups C, D, and E were decreased significantly as compared with that in group B. Emodin has a protective effect on neurons after SCI. The mechanism may be related to the activation of Nrf2-ARE pathway, reduction in the expression of NF-κB, ED-1, TNF- α, IL-1 β, and IL-6, and elevation in the expression of GFAP and NG2.
44. Inhibitory effect of extractable petroleum ether of Polyrhachis vicina Roger on neuroinflammatory response in depressed rats
Acta Pharmaceutica Sinica,Part 3: Mechanism Study,Vol 53,No. 44
The main ingredient of extractable petroleum ether of Polyrhachis vicina Roger (EPPR) is octadecene unsaturated fatty acids. Mounting evidence supports that N-3 polyunsaturated fatty acids can attenuate neuroinflammation, reduce oxidative stress, then protect neurons. In order to explore the effect of EPPR on the inflammatory response of depressed rats, the model of depression was established by chronic unpredictable mild stress (CUMS). Sucrose preference test, forced swimming test were employed to investigate the anti-depressive effect of EPPR in rat. The activation of glial cells and astrocytes in the prefrontal cortex of depressed rats was observed by immunofluorescence. The levels of inflammatory factors were measured by Quantitative Real-time PCR. NF-κB was detected by immunoblotting. EPPR could significantly improve the depressive behavior of rats, decrease NF-κB translocation to the compartment of nucleus, down-regulate the pro-inflammatory cytokines IL-1β, TNF-α and indoleamine 2, 3-dioxygenase (IDO) gene expression levels, inhibit the activation of microglia and astrocytes in depressed rats. These results suggest that EPPR could notably ameliorate inflammation induced by chronic stress, and the protective effect might be linked to the regulation of NF-κB p65.
45. Effects and Underlying Mechanism of Extract of Scutellaria baicalensis Georgi Against LPS-Induced Inflammation in BV2 Cells
Chinese Pharmaceutical Journal,Part 3: Mechanism Study,Vol 53,No. 45
OBJECTIVE To investigate the anti-inflammatory effects of Scutellaria baicalensis Georgi extract (SGE) and the underlying mechanism by using LPS-induced microglial BV2 cells. METHODS MTT assay was used to observe the cell viability. The content of NO in cell supernatant was measured using Griess reagent. The levels of IL-1β, IL-6 and TNF-α were detected by ELISA kits. The intracellular TLR4 expression was assayed by Western blotting. RESULTS The levels of NO, IL-1β, IL-6 and TNF-α were significantly increased induced by LPS in the supernatant of BV2 cells (all P < 0.01). However, treatment with 100 μg·mL ?1 SGE significantly decreased the production of related inflammatory factors including NO ( P < 0.01), IL-1β ( P < 0.01), IL-6 ( P < 0.01) and TNF-α ( P < 0.05). Furthermore, SGE significantly inhibited the TLR4 expression induced by LPS in BV2 cells. CONCLUSION SGE is able to alleviate LPS-induced inflammatory responses in BV2 cells through down-regulating the TLR4 protein expression, suggesting that SGE has therapeutic potential for treatment of neuroinflammatory diseases.
46. Polydatin attenuates airway inflammation in asthmatic mouse model via p38 MAPK/Nrf2/HO-1 pathway
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 34,No. 46
Aim To investigate whether polydatin reduces airway inflammation in asthmatic mouse model and explore whether this pathway is related to p38 MAPK/Nrf2/HO-1. Methods After the establishment of the OVA-induced asthmatic mouse model, the animals were injected with 30 mg·kg ?1 and 45 mg·kg ?1 of polydatin diluted in 0.2 mL normal saline, while the control group was replaced by normal saline. HE, PAS and Masson staining were used to observe the pathological changes of lung tissue. Diff-Quick staining was used to classify and count the number of inflammatory cells in BALF. ELISA was used to detect IgE expressions in BALF. The content of ROS in BALF cells was detected by DHR-123. The activities of antioxidant enzymes SOD, CAT and MDA in BALF were detected by the enzyme-linked immunosorbent assay kit. The expression of HO-1 in lung tissue was detected by immunohistochemistry. The protein and mRNA expressions of Nrf2 and HO-1 in lung tissue of mice were detected by Western blot and RT-PCR. Results Polydatin treatment significantly reduced inflammatory cell infiltration mucosal secretion, goblet cell proliferation and collagen deposition in the lung tissue of mice, and decreased the number of inflammatory cells and the expression of total IgE and ROS in BALF. It also increased the levels of antioxidant enzymes such as SOD and CAT, and lowered the level of MDA. Polydatin reduced the phosphorylation of p38 MAPK in the lung tissue of mice, enhanced the levels of mRNA and protein expressions of Nrf2 and HO-1 and promoted the nuclear transfer of Nrf2. The above effects of polydatin were dose-dependent. Conclusions Polydatin exerts anti-oxidative effects in OVA-induced asthmatic mouse model via anti-oxidant pathway. The mechanism may be achieved through the p38 MAPK/Nrf2/HO-1 pathway.
47. The Protective Effects of Panax notoginseng Saponins on Inflammatory Reaction of BV2 Microglial Cells Induced by LPS
Journal of Chinese Medicinal Materials,Part 3: Mechanism Study,Vol 41,No. 47
Objective: To investigate the protective effects and mechanisms of Panax notoginseng saponins (PNS) on the inflammatory reaction of BV2 microglial cells induced by lipopolysaccharides (LPS). Methods: MTT assay was used to detect the effect of different concentrations of PNS on the activity of BV2 microglia. Immunofluorescence method was used to detect the localizations and expressions of IL-1 β and NF- κB. The mRNA expressions of IL-1 β, TNF- α and iNOS were detected by PCR. The protein expressions of IL-1 β, TNF- α and COX-2 were detected by Western blot. Results: PNS at the concentrations of 6.25–50 μg/mL had no significant effect on the growth of BV2 microglial cells. Compared with the LPS model group, PNS (6.25, 12.5, 25, 50 μg/mL) could significantly inhibited the mRNA expressions of IL-1 β, TNF- α, iNOS and the protein expressions of IL-1 β, TNF- α and COX-2. The nuclear transfer and expression of IL-1 β in cytoplasm of NF- κB were significantly inhibited by 50 μg/mL PNS. Conclusion: PNS has a protective effect on the inflammatory response of BV2 microglial cells induced by LPS, which may be related to the inhibition of the activation of NF- κB pathway.
Acta Pharmaceutica Sinica,Part 3: Mechanism Study,Vol 53,No. 48
Lipopolysaccharide (LPS)-induced bone marrow-derived macrophages (BMDMs), treated with licochalcone A (LCA) and retreated with inflammasome inducers respectively (ATP and nigericin), were used to construct the inflammasome model of NLRP3 (NOD-like receptor family, pyrin domain containing 3), to investigate the inhibitory effect and the molecular mechanism of LCA on the activity of NLRP3 inflammasome. The secretion of mature interleukin (IL)-1 β, tumor necrosis factor- α (TNF- α) and caspase-1 in the supernatants were analyzed by ELISA and the Caspase-Glo ?1 inflammasome assay. Supernatants and cell lysates were analyzed for the expression of pro-caspase-1, pro-IL-1 β, ASC, NLRP3, IL-1 β, and caspase-1 by Western blot. The results showed that LCA inhibited the activity of caspase-1 and the secretion of IL-1 β, and suppressed the activity of NLRP3 inflammasome. There was also a slight inhibition on NLRC4 inflammasome induced by Lfn-Flic, but no effect on poly (dA:dT)-induced the absent in melanoma 2 (AIM2) inflammasome. Western blot results showed that LCA had no effect on the protein expression of NLRP3 and pro-IL-1 β, which was mediated by NF-κB pathway. In summary, LCA can inhibit the cleavage of pro-caspase-1 and suppress the secretion of IL-1 β to reduce the inflammation response. The study was carried out under the approval of the Scientific Investigation Board of 302 Hospital of PLA.
49. Effects of ellagic acid on inflammation and oxidative stress induced by AKT gene transfection in mice with fatty liver disease
China Journal of Chinese Materia Medica,Part 3: Mechanism Study,Vol 44,No. 49
To study the effects of ellagic acid (EA) on inflammation and oxidative stress in mice with fatty liver disease induced by AKT gene transfection. The 20 female FVB mice were randomly divided into a normal control group, a model group, as well as EA administration groups (150, and 300 mg·kg ?1·d ?1) ( n = 5). The tail vein transfection of plasmid AKT was carried out in the EA experimental groups and model group under a high pressure. Since the next day, EA was started to be administered for five successive weeks after the AKT gene transfection, while those in the model group and the normal control group were given the same amount of normal saline. After the administration, the liver tissue and serum of mice were taken. HE and oil red O staining were carried out to observe the histopathological changes in liver. The liver function was detected together with the MDA and SOD levels in the serum and liver tissue. The real-time quantitative PCR (RT-qPCR) assay was conducted to measure the mRNA expression of NF-κB and TNF- α. Western blot and immunohistochemistry were used to measure the expression of NF-κB, TNF- α and COX-2 in liver tissue. Results showed that after AKT gene transfection, the model group exhibited significant increases in the serum levels of AST and ALT. In addition, the levels of MDA in the serum and liver tissue were elevated, while the levels of SOD were decreased. Histopathological results showed obvious fatty degeneration in the liver tissue and severe damage. After EA administration, the fatty degeneration in the liver tissue was significantly alleviated. The AST, ALT, and MDA levels were lowered, while the SOD level was elevated. The expression levels of NF-κB, TNF- α, IL-6, and COX-2 were decreased in liver tissue. These results have indicated that EA had a therapeutic effect on mouse fatty liver disease induced by AKT gene transfection, and its mechanism might be related to the inhibition of NF-κB, TNF-α, IL-6, and COX-2 expression as well as the resistance against oxidant stress injury.
Chinese Pharmacological Bulletin,Part 3: Mechanism Study,Vol 35,No. 50
Aim To investigate the regulation effects and molecular mechanism of corylin on inflammasomes of NLRP3, NLRC4, and AIM2 with immortalized bone marrow-derived macrophages. Methods The effects of corylin on the activation of NLRP3, NLRC4, and AIM2 were evaluated with LPS-induced ATP, nigericin, salmonella, and poly (dA:dT). The caspase-1 activity was determined with Caspase-Glo ? 1 Inflammasome Assay. Western blot was performed to observe the protein expression levels of mature IL-1β and caspase-1 p20 in the culture supernatant, and expression levels of pro-caspase-1, pro-IL-1β, ASC, and NLRP3 in the cell lysates. Results Corylin blocked the self-slicing of pro-caspase-1 induced by ATP, nigericin, salmonella and poly (dA:dT), then suppressed the secretion of mature IL-1β mediated by caspase-1, which showed that corylin inhibited the activation of NLRP3, NLRC4, and AIM2. Moreover, corylin irreversibly attenuated the activation of NLRP3 inflammasome without affecting the NF-κB signaling pathway. Conclusions Corylin inhibits the inflammasome activation of NLRP3, NLRC4, and AIM2, and further reduces its mediated immuno-inflammatory response. Meanwhile, it provides new ideas and strategies for the treatment of immune inflammatory diseases by using corylin-related preparations.
51. Study on regulation of NLRP3/SOCS3-TLR4-NF-κB inflammatory pathway by wogonoside to improve hepatic insulin resistance
China Journal of Chinese Materia Medica,Part 3: Mechanism Study,Vol 44,No. 51
This study was to investigate the hypoglycemic effect of wogonoside to improve hepatic insulin resistance (IR) and its relative anti-inflammatory mechanism. The stable IR-HepG2 cell model was established by the combination of 1 × 10 ?9mol·L ?1insulin and 3.75 × 10 ?6mol·L ?1dexamethasone for 48 h. The changes of glucose consumption in IR-HepG2 cells with different concentrations of wogonoside (1, 5, 10, 20, and 50 μmol·L ?1) at different time points (30, 36, 48, and 54 h) were detected by glucose oxidase assay to determine the optimal onset time. Glycogen content and cell viability were respectively detected by ketone method and CCK-8 method. Cryptothermal protein 3 (NLRP3), suppressor of cytokine signaling 3 (SOCS3), Toll-like receptor 4 (TLR4), nuclear factor (NF-κB), interleukin (IL-1β), IL-6, tumor necrosis factor (TNF-α) involving in the inflammatory signaling pathway, as well as leptin, Ob-R, p-IRS2/IRS2, p-PI3K/PI3K (p85), p-Akt/Akt and glucose transporter (GLUT1/2/4) involving in the insulin signaling pathway were detected in IR-HepG2 cells by Western blot. Results showed that 20 and 50 μmol·L ?1 wogonoside significantly up-regulated the glucose consumption of IR-HepG2 cells ( P < 0.001) as compared with IR model group, and the optimal onset time was 48 h. Wogonoside had no obvious effect on the cell viability of HepG2 cells. Further studies showed that 20 and 50 μmol·L ?1wogonoside respectively increased the glycogen content of IR-HepG2 cells after 48 h treatment, especially in 50 μmol·L ?1group ( P < 0.001). Compared with IR model group, wogonoside not only inhibited the protein expression of inflammatory nuclear transcriptional factors NLRP3, SOCS3, TLR4, NF-κB, but also decreased the expression of downstream inflammatory effect factors IL-1β, IL-6 and TNF-α. In addition, wogonoside elevated Ob-R, p-IRS2/IRS2, p-PI3K/PI3K (p85), p-Akt/Akt and GLUT1/2/4 protein expression, whereas it suppressed leptin expression that was regulated by SOCS3. Wogonoside could promote glucose uptake and increase glycogen content to enhance insulin sensitivity in IR-HepG2 cells. The hypoglycemic effect may be related to the intervention of NLRP3/SOCS3-TLR4-NF-κB inflammatory pathway and decrease of inflammatory factor expression.
52. Investigation on the inflammation network mechanisms of Wutou decoction acting on neuropathic pain
Acta Pharmaceutica Sinica,Part 3: Mechanism Study,Vol 54,No. 52
Wu-tou decoction (WTD) was originally recorded in the Synopsis of the Golden Chamber and it had been widely used for the treatment of neuropathic pain (NP) with exact therapeutic efficacy. However, the underlying molecular mechanisms still remain unclarified. Thus, in this research, we aimed at clarifying the underlying molecular mechanisms of WTD against NP by combining network analysis and experimental validation based on the spinal nerve ligation (SNL) model. Firstly, the network analysis indicated that key targets of WTD were significantly involved in the MAPK signaling pathway ( P = 4.04 E-12) and four important components of the above pathway, AKT kinase (AKT), MAP kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK) and transcription factor AP-1 (JUN) had been reported to play a vital role in neuroinflammation during the disease process of NP. Then, experimental validation results proved that WTD markedly reduce the severity of mechanical allodynia ( P < 0.01) and cold hypersensitivity ( P < 0.05) of SNL rats. In addition, Western blot results provided evidence that the phosphorylated protein expression levels of AKT, MKK4, JNK and JUN in the superficial lamina of spinal cord of SNL rats were markedly increased ( P < 0.001), and WTD could improve the phosphorylated protein expression level of AKT ( P < 0.001) which was reported to be nerve protective and attenuate the phosphorylated protein expression levels of MKK4, JNK and JUN ( P < 0.01) which were closely involved in neuroinflammation. In conclusion, this study indicated that WTD might exert anti-hyperalgesia action through the inhibition of neuroin‐flammation mediated by AKT-MKK4-JNK-JUN in the MAPK signaling pathway. These findings also provided scientific evidences that WTD might be a promising candidate for NP. Animal experiments in this study were approved by the Ethics Committee of Experimental Animals of the China Academy of Chinese Medical Sciences.
53. Mechanism of Danzhi Jiangtang Capsules in treatment of diabetic macroangiopathy in GK rats based on correlation between microRNAs and inflammatory factors
China Journal of Chinese Materia Medica,Part 3: Mechanism Study,Vol 44,No. 53
This study was aimed to investigate the mechanism of Danzhi Jiangtang Capsules (DJC) in the treatment of diabetic macroangiopathy in Goto-Kakizaki (GK) rats. The diabetic macroangiopathy rat model was induced by feeding high-fat and high-sugar combined with endothelial nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) (0.1 g·L ?1·d ?1). According to the random number table, the model rats were randomly divided into the model group, DJC groups (1 260, 630, and 320mg·kg ?1), atorvastatin group (105 mg·kg ?1) and metformin group (10 mg·kg ?1), with 12 rats in each group. The rats received intragastric administration for eight weeks. Twelve Wistar rats were selected as the normal control. The changes in the body weight, water intake, blood glucose, plasma total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), interleukin (IL-1 β), IL-6, tumor necrosis factor (TNF- α), nitric oxide (NO), endothelin (ET-1) were observed in these rats. Aortic tissue was taken and the pathological changes were observed by HE staining. RT-PCR was used to detect the mRNA levels of IL-1 β, IL-6, and TNF- α in rat aorta. The stem loop RT-PCR was conducted to detect the levels of miRNA-126, miRNA-155, miRNA-146a, and miRNA-21 in rat plasma and aortic tissue. The canonical correlation between miRNAs and inflammatory factors was then analyzed. The results showed that DJC increased the rat body weight, lowered water intake, reduced the random blood glucose, reversed the rat aorta tissue damage, reduced serum TC, TG, LDL-C, ET-1, IL-1 β, IL-6, and TNF- α, as well as miRNA-155, miRNA-146a and miRNA-21 levels in serum, elevated plasma HDL-C, NO content, reduced the aorta mRNA of IL-1 β, IL-6, TNF- α, and the miRNA-155, miRNA-146a and miRNA-21, elevated miRNA-126 expression in aorta. Aortic miRNA-126, miRNA-155, miRNA-146a and miRNA-21 expression levels were typically correlated with the expression of inflammatory factors, among which miRNA-126 was negatively correlated, miRNA-155, miRNA-146a and miRNA-21 were positively correlated with the factors. These results suggested that DJC had therapeutic effects on diabetic vascular complications, and the mechanism of action might be related to its regulation of miR-NA-126, miRNA-155, miRNA-146a and miRNA-21 levels, as well as the reduction of inflammatory factors and alleviation of vascular inflammatory response.
54. Anti-inflammatory Effect and Mechanisms of Flemingia macrophylla Merr. for Chronic Pelvic Inflammatory Disease in Rats
Chinese Pharmaceutical Journal,Part 3: Mechanism Study,Vol 54,No. 54
OBJECTIVE To explore the anti-inflammatory effect and mechanism of Flemingia macrophylla Merr. on chronic pelvic inflammatory disease (CPID) rats through in vivo and in vitro experiments. METHODS LPS-stimulated mouse macrophages RAW264.7 in vitro were applied to observe the effects of F. macrophylla Merr. on expressions of inflammatory factors (TNF-α, and IL-6) and free radical (NO) and inflammation related proteins (IκBα, and iNOS). Estradiol was used to induce in vivo pseudo-oestrous cycle, and then the CPID rat model was established by the infection of the mixture of Staphylococcus aureus, Escherichia coli and Ureaplasma urealyticum via transvaginal implantation into the uterine cavity. After successful modeling, the rats were randomly divided into the model group, the positive drug group (Fuyankang), the low-dose group (4.25 g·kg ?1), the medium-dose group (8.5 g·kg ?1), and the high-dose group (17 g·kg ?1), and the treatment was given by gavage continuously for 21 d. The rats in the normal group, sham operation group and mode group were given saline solution. After the final administration, rats’ blood was collected from orbital venous plexus, and the serum levels of inflammatory cytokines IL-6 and TNF-α were measured by Elisa method. The rat’s uterus and its accessories were stained with HE to observe the morphologies of endometrium and fallopian tubes. RESULTS F. macrophylla Merr. could reduce the release levels of TNF-α, IL-6 and NO, inhibit the degradation of IκBα, and reduce the expression of inducible nitric oxide synthase (iNOS) in RAW264.7 cells induced by LPS. Moreover, it could significantly improve the pathological state of the uterus and fallopian tubes of rats with CPID, and reduce TNF-α and IL-6 in the serum of rats with CPID. CONCLUSION The mechanism of F. macrophylla Merr. in inhibiting CPID may be related to the inhibition of IκBα degradation, the reduced release of inflammatory factors such as TNF-α and IL-6, the down-regulation of iNOS, as well as the decreased generation of NO.
55. In vitro antitumor and anti-inflammatory activities of triterpenoids with different polarity in the fruiting bodies of Ganoderma lingzhi
Mycosystema,Part 3: Mechanism Study,Vol 38,No. 55
To investigate the activities of triterpenoids with different polarity from the fruiting body of Ganoderma lingzhi, the total triterpenoid extract was separated into different-polarity extracts by macroporous resin. Fingerprints and the content of triterpenoids were analyzed by HPLC. Antitumor and anti-inflammatory activities of different-polarity extracts were compared. The results showed that the total triterpenoids can be separated by D-101 macroporous resin effectively into two polar parts, medium polar triterpenoids and low polar triterpenoids, with the content of (279.00 ± 2.90) mg/g and (94.52 ± 2.03) mg/g respectively. The antitumor activity of low polar triterpenoids is much stronger than that of medium polar triterpenoids. The IC 50 of inhibition to K562 cells, SW620 cells, and L1210 cells are (74.12 ± 1.94) μg/mL, (121.45 ± 2.13) μg/mL and (13.52 ± 1.13) μg/mL, respectively. The inhibitory effect of low polar triterpenoids on PMA-stimulated respiratory burst in RAW264.7 macrophages is also obviously stronger than that of medium polar triterpenoids. The comparison of the inhibitory effect between different triterpene monomers on PMA-stimulated respiratory burst in RAW264.7 macrophages shows that ganoderenic acid F, ganoderic acid DM and ganoderol B, with low polarity, have stronger anti-inflammatory activity than ganoderic acid C 2 and ganoderic acid A with medium polarity. It is proved that the in vitro antitumor and anti-inflammatory activities of triterpenoids with different polarity from the fruiting bodies of Ganoderma lingzhi are different significantly. This study provides reference for improving quality standards of the Ganoderma lingzhi products.
56. Spectrum-effect relationship of anti-inflammatory activity of Shufeng Jiedu Capsule based on neural network analysis
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 50,No. 56
Objective To establish the spectrum-effect relationship network model of Shufeng Jiedu Capsule for anti-inflammatory activity and explore potential effective compounds of its anti-inflammatory effects. Methods On the basis of the establishment of fingerprint analysis method for Shufeng Jiedu Capsule, the uniform design method was used to divide eight herbs in Shufeng Jiedu Capsule into groups of different proportions to determine their inhibition ratios of TNF-α and IL-6 released by LPS-induced RAW264.7 cells. Pharmacodynamic data and chemical information of MS fingerprints of each group were analyzed with the BP artificial neural network to get the anti-inflammatory effect of each peak to establish the chromatography-efficacy relationship. Results According to the optimal neural network model, 14 characteristic peaks were calculated and found to be significantly correlated with the anti-inflammatory activity of Shufeng Jiedu Capsule. Conclusion Through the spectrum-effect study, it is speculated that the anti-inflammatory components of Shufeng Jiedu Capsule may be the 14 compounds including rhein, emodin, verbenalin, verbascoside, pinoresinol-β- D-glucoside, forsythoside A, polydatin, etc., which provides a reference for the quality control and determination of effective components.
Chinese Traditional and Herbal Drugs,Part 3: Mechanism Study,Vol 50,No. 57
Objective To study the antipyretic and anti-inflammatory constituents from the active fraction of Reduning (RDN) Injection. Methods In this study, the active fraction of RDN Injection was screened by the LPS-induced mouse endotoxin shock model. The chemical constituents were isolated by chromatography on HP-20 macroporous resin, silica gel, MCI, ODS, reverse MPLC, and HPLC repeatedly, and their structures were identified by spectral data and physicochemical property. Taking PGE 2 as the evaluating indicator, the model of LPS-induced RAW264.7 cells was used to evaluate the in vitro anti-inflammatory activity of these compounds. Results The 95% ethanol eluate of RDN Injection on the macroporous adsorption resin column was proved to be the antipyretic and anti-inflammatory active fraction of RDN Injection. A total of 24 compounds were isolated and identified as (4a S,7a S,7b S)-4,4a,7a,7b-tetrahydro-2 H-1,7-dioxacyclopent [cd] indene-5-carboxylic acid methyl ester (1), (4a S,7a S)-1,4a,5,7a-tetrahydro-7-(hydroxymethyl)-cyclopenta [c] pyran-4-carboxy licacid methyl ester (2), 3α,5α-tetrahydrodeoxycordifoline lactam (3), R-( Z)-4-methyl-5-[(2′,6′,6′-trimethyl-4′-oxo-2′-cyclohexen-1′-yl) methylene]-2(5 H)-furanone (4), (1α,2α,3β,4β)-2,4-bis(4-hydroxy-3-methoxyphenyl)-1,3-cyclobutanedicarboxylic acid (5), 4-[(6- O-benzoyl-β- D-glucopyranosyl) oxy]-3-methoxybenzoic acid (6), syringaresinol (7), E-3-(3,4-dihydroxybenzylidene)-5-(3,4-dihydroxyphenyl) dihydrofuran-2-one (8), 6,7-dimethoxy coumarin (9), 7-hydroxy-6-methoxy coumarin (10), salicylic acid (11), syringaldehyde (12), phenylacetic acid (13), vanillin (14), caffeic acid (15), aceto-vanillone (16), 3,5-di- O-caffeoylquinic acid (17), 4,5-di- O-caffeoylquinic acid (18), 3,4-di- O-caffeoylquinic acid methyl ester (19), 3,5-di- O-caffeoylquinic acid methyl ester (20), 4,5-di- O-caffeoylquinic acid methyl ester (21), 5- O-caffeoylquinic acid ethyl ester (22), 3,5-di- O-caffeoylquinic acid ethyl ester (23), and 4,5-di- O-caffeoylquinic acid ethyl ester (24). Among them, compounds 1, 10, and 14–24 significantly inhibited PGE 2 expression in RAW 246.7 cells stimulated by LPS. Conclusion Compounds 1–9, 11–13, and 22–24 are isolated from RDN Injection for the first time; organic acids may be one of the main pharmacodynamic substances of RDN injection for antipyresis and anti-inflammation.
58. Analysis on transcriptionomic characteristics of Naoxintong Capsules in prevention of post-ischemic inflammation based on RNA-Seq technology
China Journal of Chinese Materia Medica,Part 3: Mechanism Study,Vol 45,No. 58
In this research, high-throughput sequencing was used to investigate the mechanism of Naoxintong Capsules (NXTC) in prevention of post-ischemic inflammation. First, microglia BV-2 inflammatory model was induced by 1.0 μg·mL ?1 LPS to investigate the effects of the intestinal absorption solutions of NXTC (NXTCIA) at different concentration (62.5, 31.25, 15.63, 7.81 μg·mL -1) on LPS-induced BV-2 inflammatory factors in microglia. Then, the RNA-Seq high-throughput sequencing method was adopted to identify the differentially-expressed mRNAs in microglia BV-2 after pre-treatment with NXTC. GO and KEGG enrichment analysis was used to screen the potential biological processes and related signaling pathways of NXTC in inhibiting inflammation. The results showed that four NXTCIA concentration gradients could significantly inhibit the release of LPS-induced inflammatory mediators in BV-2 in a dose-dependent manner. Furthermore, the high-throughput sequencing results showed that 392 mRNA transcripts were reversed following pre-treatment with NXTC. The GO enrichment analysis showed that the transcripts reversed by NXTC were mainly involved in the Toll-like receptor signaling pathway, chemokine signaling pathway, and TNF signaling pathway. In summary, our findings showed that NXTC treatment could provide protective effects against inflammatory response and the mechanism might be related to the regulation of the Toll-like receptor signaling pathway, chemokine signaling pathway, and TNF signaling pathway.
59. Assessment of antioxidant and anti-inflammatory potential of the water extract of Floccularia luteovirens in diabetic rats
Mycosystema,Part 3: Mechanism Study,Vol 38,No. 59
Oxidative stress is thought to be a key risk factor in the development of diabetes mellitus, tumor and hepatic diseases. Blocking or retarding the reactions of oxidation and the inflammatory process by antioxidants could be a promising therapeutic intervention for prevention or treatment of diabetes mellitus. Floccularia luteovirens, previously interpreted as Armillaria luteovirens, as a precious edible-medicinal fungus, is widely distributed in the Qinghai-T ibet Plateau. Its fruiting body has many pharmacological effects such as anti-tumor, anti-inflammation and antioxidant. In this study, we investigated the anti-oxidant and anti-inflammatory effects of the water extract of Floccularia luteovirens (FLW) on STZ-induced type II diabetes mellitus in rats. After treatment with metformin and FLW for 8 weeks, the results showed that FLW improved glucose tolerance and promoted exogenous glucose consumption. MDA and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) levels were reduced and SOD level was advanced by FLW in serum. Taken together, FLW showed antioxidative and anti-inflammatory effects in diabetic rats by inhibiting oxidative stress and the inflammation.