Publisher(s): China Academic Journals (CD Edition) Electronic Publishing House Co., Ltd.
ISBN: ISBN 978-7-499-00976-9 pdf
First Published: 2020.11.23
Discipline(s): Medicine & Public Health
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Arthritis (TCM), as part of China’s Medicine Progress Series, has 53 excellent articles regarding arthritis management with TCM. The articles included are recommended by the editorial boards of journals in which they were originally published, and the content mainly covers mechanism research, clinical research, and experience discussion, fully demonstrating the unique therapies and excellent efficacy of TCM in arthritis management, with scientific methodological support and objective conclusions. The original articles were published in Chinese and this book is a compilation of English version of the selected articles.
Doctor WU Zhongchao is a doctoral supervisor, a chief physician, a consultant expert of the Central Health Protection Commission, one of the fifth batch of the national distinguished veteran doctors of TCM, and the chairman of the Specialized Committee on
1. Anti-inflammatory and synovial-opioid system effects of electroacupuncture intervention on chronic pain in arthritic rats
Chinese Acupuncture & Moxibustion,Part 1: Mechanism Study,Vol 35,No. 01
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.
2. Effect of Electroacupuncture on Serum TNF-α, IL-1β and Intercellular adhesion molecule 1 Levels in Rheumatoid Arthritis Rats
Acupuncture Research,Part 1: Mechanism Study,Vol 41,No. 02
Objective To investigate the effects of electroacupuncture on the contents of serum tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and intercellular cell adhesion molecule-1 (ICAM-1) of a model of rats with collagen induced arthritis and to discuss the mechanism by which electroacupuncture treats rheumatoid arthritis (RA). Methods A total of 60 Wistar male rats were included in the current study, where 8 rats were randomly selected as a normal group, while others were adopted to establish an RA model using chicken type II collagen and Freund’s adjuvant incomplete. After successful modeling, 24 rats were randomly divided into three groups ( n = 8): a model group, a prednisone group and an electroacupuncture group. Needles were inserted into Zusanli and Kunlun points on both sides of rats of the electroacupunture group, 30 min per time daily for 10 consecutive days. Those in the prednisone group were intragastrically administrated with prednisolone acetate (0.1 mL/10 g). Then, the diameters of the left ankle joint were measured before and after modeling, and after treatment. The contents of serum TNF-α, IL-1β and ICAM-1 were examined by ELISA. Results Compared with the normal group, rats in the model group had larger ankle diameters and higher contents of serum TNF-α, IL-1 b and ICAM-1 ( P < 0.05). Compared with the model group, reduced ankle diameters and decreased contents of serum TNF-α, IL-1 b and ICAM-1 were found in rats after electroacupunture or prednisone treatment ( P < 0.05). No statistical difference was seen between the electroacupunture and prednisone groups ( P > 0.05). Conclusion Electroacupunture is effective to treat rats with RA, which may be associated with its abilities to effectively weaken the secretion of serum TNF-α, IL-1 b and ICAM-1.
3. Effects of Chronological Moxibustion on Circadian Rhythm Activities of Hypothalamus-Pituitary-Axis in Rheumatoid Arthritis Rats
Acupuncture Research,Part 1: Mechanism Study,Vol 41,No. 03
Objective To observe the effect of chronological moxibustion on plasma corticosterone (CORT), corticotrophin releasing factor (CRF) and adrenocorticotropic hormone (ACTH) levels in rheumatoid arthritis (RA) rats, so as to explore its mechanisms underlying improvement of RA. Methods A total of 144 SD rats were randomly divided into normal control, model, moxibustion, sham-adrenalectomy (ADX, sham-ADX), ADX and ADX + moxibustion groups which were further separately divided into 4 subgroups (4 time-points: 12 pm, 6 am, 12 am, 6 pm, n = 6/subgroup). The RA model was established by subcutaneous injection of Freund's complete adjuvant (FCA, 0.1 mL) into the right footpad (for rats of the latter 5 groups). In rats of the ADX and ADX + moxibustion groups, bilateral adrenal glands were removed under anesthesia. Moxibustion was applied to unilateral Shenshu (BL 23) and Zusanli (ST 36) for 10 min, once daily for 6 days in a week, and continuously for 3 weeks. The paw swelling volume was measured and plasma CORT, ACTH and CRF contents were assayed by ELISA and the circadian rhythm was analyzed by cosine curve fitting (cosinor) method. Results The paw swelling volume was significantly increased in the model group than in the normal control group ( P < 0.05), and decreased remarkably in the moxibustion group ( P < 0.05). The paw volume was markedly increased in the ADX + moxibustion group in comparison with the moxibustion group ( P < 0.05). In intact rats, plasma CRF and ACTH contents were significantly increased ( P < 0.05), and plasma CORT was obviously decreased in the model group ( P < 0.05). After moxibustion, the increased plasma CRF and ACTH contents and the decreased plasma CORT level were considerably reversed ( P < 0.05). In ADX rats, plasma CRF, ACTH and CORT contents of ADX and ADX + moxibustion groups had no significant changes compared with the sham-ADX group ( P > 0.05). Results of the cosine curve fitting analysis showed that the peak phases of plasma CORT contents were ?355.78° at about 23:43 in the normal control group, ?309.05° at about 20:36 in the model group, ?326.50° at about 21:46 in the moxibustion group, ?291.65° at about 19:27 in the ADX group and ?300.87° at about 20:31 in the ADX + moxibustion group. The peak phases of plasma ACTH contents were ?324.08° at about 21:37 in the normal control group, ?295.39° at about 19:41 in the model group, ?310.81° at about 20:43 in the moxibustion group, ?146.51° at about 9:46 in the ADX group and ?267.64° at about 17:50 in the ADX + moxibustion group. The peak phases of plasma CRF contents were ?257.47° at about 17:10 in the normal control group, ?184.74° at about 12:19 in the model group, ?263.00° at about 17:32 in the moxibustion group, ?202.46° at about 13:30 in the ADX group and ?232.84° at about 15:31 in the ADX + moxibustion group. It suggests that in the intact RA rats, moxibustion intervention may inhibit modeling-induced decrease of the circadian rhythm of plasma CORT, and hyperactivity of ACTH and CRF circadian rhythm. Conclusion Moxibustion intervention can relieve paw swelling in RA rats and modulate the circadian rhythm activities of plasma CORT, ACTH and CRF levels (HPA axis activities) in intact animals.
4. Effects of Short Thrust Needing plus Electroacupuncture Intervention on Cartilage Tissue in Rabbits with Knee Osteoarthritis
Acupuncture Research,Part 1: Mechanism Study,Vol 41,No. 04
Objectives: to observe the effectiveness of short thrust needling (STN, close-to-bone needing) plus electroacupuncture (EA) in healing knee cartilage tissue and in regulating expressions of cartilage vitamin K dependent gamma-glutamyl carboxylase (GGCX), matrix metalloproteinase-13 (MMP 13) and serum uncarboxylated matrix gla protein (ucMGP) in rabbits with knee osteoarthritis (KOA), so as to reveal its mechanism underlying improvement of KOA. Methods: Forty New Zealand rabbits were randomly divided into Normal, Model, EA and STN+EA groups ( n = 10 in each group). The KOA model was created by cutting the medial lateral ligament and medial parapatellar arthrotomy of rabbits as described by Hulth and colleagues. For rabbits in the STN+EA group, “Neixiyan” (EX-LE 4) and “Waixiyan” (ST 35) were punctured with filiform needles by controlling the needle-tip obliquely to advance till the bone surface of the knee joint cavity, and “Yinlingquan” (SP 9) and “Zusanli” (ST 36) punctured by holding the filiform needles vertically along the tibia, and “Liangqiu” (ST 34) was punctured by controlling the filiform needle to advance till the thigh-bone, followed by EA stimulation. EA (2Hz/100 Hz, 1–3 mA) was applied to unilateral EX-LE 4 and ST 35, and ST 36 and SP 9, separately for 20 minutes, once daily for 20 days except weekends. The pathological changes of the knee cartilage cells were observed by using H.E. staining, Toluidine blue staining and electron transmission microscope, respectively. The immunoactivity of GGCX of the knee cartilage was determined by immunohistochemistry and the expression levels of GGCX and MMP 13 proteins in the cartilage were detected by Western blot, and the content of serum ucMGP was assayed by ELISA. Results: H.E. staining, Toluidine blue staining and electron transmission microscope results showed that pathological changes of knee cartilage cells in structure after modeling were improved in both the STN + EA and EA groups, particularly the former group. In comparison with the Normal group, the expression levels of GGCX proteins in the cartilage tissue showed by both Western blot and immunohistochemistry were notably down-regulated ( P < 0.01), and the cartilage MMP 13 protein expression and serum ucMGP content were considerably up-regulated in the Model group ( P < 0.01, P < 0.05). After STN + EA and simple EA, the decreased GGCX and the increased MMP 13 expression and serum ucMGP content were reversed ( P < 0.01, P < 0.05). The effects of STN + EA were significantly superior to those of simple EA in down-regulating MMP13 and ucGLA levels, and up-regulating GGCX expression. Conclusion: Both STN + EA and simple EA can effectively improve pathological changes of cartilage cells in KOA rabbits, which may be associated with their actions in up-regulating the expression of cartilage GGCX proteins and lowering the levels of serum ucMGP content and cartilage MMP 13 protein expression, and the effects of STN + EA are better.
5. Effect of Warm-needle-moxibustion on Behavior Reactions and TNF-α and MMP-3 Contents in Knee Cartilage of Rabbits with Knee Osteoarthritis
Acupuncture Research,Part 1: Mechanism Study,Vol 41,No. 05
Objective To observe the effect of warm-needle-moxibustion on behavior changes and the levels of tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-3 (MMP-3) in the knee cartilage tissue of rabbits with knee osteoarthritis (KOA) so as to explore its mechanisms in relieving KOA. Methods Thirty male Japanese white rabbits were randomly divided into normal, model, and treatment groups ( n = 10 in each group). KOA model was established by using immobilization method. The internal and external “Dubi” (ST 35), “Xuehai” (SP 10) and “Yanglingquan” (GB 34) were punctured with filiform needles first, followed by attaching an ignited moxa cone to the handle of each inserted needles. The treatment was conducted for 20 min every time, once daily for 4 weeks. The behavioral scores were assessed according to the pain severity, gait, joint motion range and articular swelling severity in reference to the modified Lequesne's methods. The content of TNF-α and MMP-3 in the knee joint cartilage tissue was assayed by radioimmunoassay, and pathological changes of the articular cartilage tissue were observed under microscope after sectioning and H.E. staining. Results The Lequesne total score and the cartilage TNF-α and MMP-3 content were significantly increased in the model group than in the normal group ( P < 0.05), and obviously down-regulated after warm-needle-moxibustion intervention ( P < 0.05). Under light microscope, the inflammatory infiltration of the articular cartilage and the disordered arrangement of chondrocytes in KOA rabbits were improved in the treatment group. Conclusion Warm-needle-moxibustion intervention can improve the motor function of rabbits with KOA, which may be related to its effects in reducing the level of TNF-α and MMP-3 in articular cartilage tissue.
6. Repair effects of close-to-bone needing combined with electroacupuncture on extracellular matrix of cartilage in rabbits with knee osteoarthritis
Chinese Acupuncture & Moxibustion,Part 1: Mechanism Study,Vol 36,No. 06
Objective To observe the effects of close-to-bone needing combined with electroacupuncture (EA) on cartilage collagen type Ⅱ/discoidin domain receptor/matrix metalloproteinase 13 (collagen type Ⅱ/DDR2/MMP 13) signaling pathway in rabbits with knee osteoarthritis (KOA), and to explore the possible action mechanism of this method on repair of extracellular matrix of knee cartilage. Methods Forty New Zealand white rabbits were randomly assigned into a normal group (10 rabbits) and a model establishing group (30 rabbits). The HulthTelhag technique was applied to establish the model of KOA, and X-ray was used for outcome assessment. The rabbits with successful modeling were randomly assigned into a model group, a close-to-bone needing group, a regular acupuncture group, with 10 rabbits in each one. The rabbits in the close-to-bone needing group were treated with close-to-bone needing and EA. The rabbits in the regular acupuncture group were treated with regular acupuncture and EA. “Neixiyan” (EX-LE 4), “Dubi” (ST 35), “Yinlingquan” (SP 9), “Zusanli” (ST 36), and “Liangqiu” (ST 34) were selected in the two groups. The intervention was given for 20 min per time, once a day, five days as a course, with an interval of two days between courses, for a total of four courses. Rabbits in normal and model group were immobilized without any treatment. After the treatment, western blotting method was applied to evaluate the expression of DDR2 and collagen type Ⅱ; The activities of collagen type Ⅱ, DDR2 and MMP 13 were assessed by immunohistochemistry method; The mRNA expression levels of DDR2and MMP 13 were determined by reverse transcription-polymerase chain reaction (RT-PCR). Results Compared with the normal group, the activities of collagen type Ⅱ were significantly reduced in the other groups (all P < 0.01), while the activity and mRNA expression levels of DDR2 and MMP 13 were notably increased (all P < 0.01). Compared with the model group, the activities of collagen type Ⅱ in the close-to-bone needing group and regular acupuncture group were increased (both P < 0.01), while the activity and mRNA expression levels of DDR2 and MMP 13 were reduced (all P < 0.01). Compared with the regular acupuncture group, the activity and mRNA expression levels of MMP 13 and DDR2 in the close-to-bone needing group were reduced (all P < 0.01), while the activities of collagen type Ⅱ were increased ( P < 0.01). Conclusion The close-to-bone needing combined with EA and regular EA could both promote the repair of knee cartilage, whereas, closing-to-bone needing combined with EA shows a superior efficacy. The mechanism might be associated with the blocking effect of collagen type Ⅱ/DDR2/MMP13 signaling pathway and the inhibiting effect of degradation in extracellular matrix of cartilage.
Chinese Acupuncture & Moxibustion,Part 1: Mechanism Study,Vol 37,No. 07
Objective To compare the effects of different acupuncture methods on urine metabolites in rheumatoid arthritis (RA) rabbits, and to explore the specificity mechanism of heat-reinforcing acupuncture for RA. Methods A total of 40 clean purple-blue rabbits were randomly allocated to a normal group, a model group, a mild reinforcing-reducing needling (MRRN) group, a twirling-reinforcing needling (TRN) group and a heat-reinforcing needling (HRN) group, 8 rabbits in each one. Except the normal group, the rabbits in the remaining groups were treated with ovalbumin and freezing to establish RA model. The rabbits in the MRRN group, TRN group and HRN group were treated with MRRN, TRN and HRN at “Zusanli”(ST 36), respectively, 30 min per treatment, once a day for seven days. After treatment, 24-h urine was collected. The rabbits were sacrificed to collect synovial tissues of knee to perform morphology observation; the liquid chromatography quadrupole time-offlight mass spectrometry (LC-Q/TOF-MS) was applied to measure urine metabolites. All the data were analyzed by principal component analysis (PCA)and partial least squares discriminant analysis (PLS-DA). Results Compared with the normal group, the leucine-related metabolites, as main urine metabolites, were decreased in the model group ( P < 0.05), while the purine-related metabolites and tryptophan-related metabolites were increased ( P < 0.05). Compared with the model group, the leucine-related metabolites, as main urine metabolites, were increased in the three needling groups after treatment ( P < 0.05), while the tryptophan-related metabolites and purine-related metabolites were decreased ( P < 0.05), moreover, the leucine-related metabolites in the HRN group were obviously higher than those in the MRRN group and TRN group ( P < 0.05). Conclusion MRRN, TRN and HRN can regulate the pathway of leucine metabolism (energy metabolism), purine metabolism (oxidative damage) and tryptophan metabolism (immune regulation) for RA, The specificity of HRN for RA focuses on regulation of leucine metabolism (energy metabolism).
8. Effects of moxibustion on Treg/Th17 cell and its signal pathway in mice with rheumatoid arthritis
Chinese Acupuncture & Moxibustion,Part 1: Mechanism Study,Vol 37,No. 08
Objective To observe the effects of moxibustion on Treg/Th17 imbalance and related signal pathway in mice with rheumatoid arthritis (RA), so as to explore the action mechanism of moxibustion on RA. Methods Twenty-four DBA/1J male mice were randomly divided into a normal group, a model group, a sham moxibustion group and a moxibustion group, with six mice in each one. RA model was induced by subcutaneous injection of type II collagen and adjuvant at tail in mice in all groups other than the normal group. The mice in the moxibustion group were treated with moxibustion at “Zusanli” (ST36) and “Shenshu” (BL23), 1 mg per cone, six cones per acupoint. The consecutive six-day treatment was taken as one course, and totally two courses were given with an interval of 2 d between courses. The mice in the sham moxibustion group were treated with immobilization as the moxibustion group. The effects of moxibustion on joint swelling was evaluated by RA scale of collagen induced arthritis (CIA); The pathological changes of joint inflammation were observed by HE staining; The cell count of Th17 and Treg in spleen was analyzed by flow cytometry; The contents of cytokine IL-1β, IL-6, IL-10, IL-17, IL-23, TGF-β and Galectin-9 were analyzed by ELISA; The mRNA and protein expression levels of Foxp3, Galectin-9, RORγt, CARMA1, NF-κB were analyzed by Real-time PCR and Western Blotting method. Results During days 10–12 after the secondary immune, redness and swelling of one or two ankle joints, soles and toe joints were observed, indicating the successful establishment of RA model. On day 15 of moxibustion treatment, the joint swelling was improved in the moxibustion group and the sham moxibustion group, with better results noticed in the moxibustion group ( P < 0.05). As for pathological changes, compared with the normal group, the articular surface became rough and the synovial layer became thinner in the model group, which were recovered to a certain extent in the sham moxibustion group; The articular surface was smooth and synovial layer was thicker in the moxibustion group, which were similar to those in the normal group. The results of flow cytometric test indicated that the cell count of Treg in the model group was reduced but that of Th17 was increased as compared with those in the normal group (both P < 0.01); The moxibustion could increase significantly the cell count of Treg ( P < 0.05), but no effect was observed on Th17 ( P > 0.05). The results of ELISA test indicated that the differences in elevation of IL-1β, IL-6, IL-17, IL-23, TGF-β as well as in the reduction of IL-10 were not significant between the sham moxibustion group and the moxibustion group (all P > 0.05); Moxibustion treatment could increase the content of Galectin-9 which was reduced in RA mice ( P < 0.05). The results of RT-PCR and Western blotting test indicated that the mRNA and protein expression levels of Foxp3 and Galectin-9 were reduced in the model group (all P < 0.01), which could be up-regulated by moxibustion treatment ( P < 0.05, P < 0.01); The mRNA and protein expression levels of RORγt, CARMA1, and NF-κB were increased (all P < 0.01), which could be down-regulated by moxibustion treatment ( P < 0.05, P < 0.01). Conclusion Moxibustion can improve the swelling of joint and inflammatory reaction of joint synovial in RA mice; The mechanism may be related to the regulation of Treg cell number in spleen as well as the mRNA and protein expression of Foxp3, Galectin-9, RORγt, CARMA1, and NF-κB.
Journal of Chinese Medicinal Materials,Part 1: Mechanism Study,Vol 38,No. 09
Objective: To explore the changes of intestinal mucosal immunity in collagen-induced arthritis rats and the impact of madecassoside on these changes. Methods: Collagen-induced arthritis was established in female Wistar rats. Treatment group was orally administrated madecassoside once daily for consecutive 21 days, while blank control and model groups were orally administered saline at the same volume. The concentrations of s Ig A in small intestine content and IFN-γ in small intestinal tissue homogenate were determined by ELISA. The proportions of CD4+T and CD8+T in the epithelium and laminar propria of small intestine were detected by flow cytometry, and the ratios of CD4+/ CD8+were calculated. The relative expressions of CD80, CD86, IL-6, IL-12 and Foxp3 mRNA in the small intestine were determined by real-time PCR. Results: Compared with blank control rats, the concentrations of s Ig A in small intestine content and IFN-γ in small intestinal tissue homogenate from model rats were increased, the ratios of CD4+/ CD8+in the epithelium and laminar propria of small intestine were higher and the relative expressions of CD80, CD86, IL-6 and IL-12 mRNA in the small intestine were increased. Madecassoside treatment decreased the concentrations of s Ig A in small intestine content and IFN-γ in small intestinal tissue, downregulated the ratios of CD4+/ CD8+in the epithelium and laminar propria and decreased the relative expressions of CD80, CD86, IL-6 and IL-12 mRNA, while upregulated the relative expression of Foxp3 mRNA in the small intestine. Conclusion: The intestinal mucosal immune response is enhanced in collagen-induced arthritis rats, the antigen presenting cells are activated abnormally and the immune tolerance is disturbed. Madecassoside treatment can downregulate the intestinal mucosal immune response and benefit for the induction and maintenance of intestinal immune tolerance.
10. Effect of Jinwu Jiangu Recipe on Expressions of NF-κB and IL-17 in Collagen Induced Arthritis Model Rats
Chinese Journal of Integrated Traditional and Western Medicine,Part 1: Mechanism Study,Vol 36,No. 10
Objective To explore the effect of Jinwu Jiangu Recipe (JJR) on the expression of synovial cells′ nuclear factor-kappa B (NF-κB) and serum interleukin 17 (IL-17) in collagen induced arthritis (CIA) rats. Methods Totally 60 Wistar rats were randomly divided into 6 groups, i.e., the blank control group, the model group, high, middle, and low dose JJR treatment groups, and the tripterygium control group, 10 in each group. Except for rats in the blank control group, CIA model was established in rats of the rest 5 groups. Then they were treated from the 7 th day of modeling. After 4 weeks of medication they were sacrificed, serum collected, and synovium of joints were isolated. The expression of serum IL-17 was detected in synovium of joints by enzyme linked immunosorbent assay (ELISA). The expression of NF-κB/P65, IκBα and NF-κB/P50 were detected by Western blot. Results Compared with the blank control group, the serum IL-17 level increased in the model group ( P < 0.01). Compared with the model group, the serum IL-17 level obviously decreased in high and middle dose JJR groups and the tripterygium control group ( P < 0.01). Results of Western blot showed, when compared with the blank control group, protein activities of NF-κB/P65 and NF-κB/P50 were significantly enhanced in the model group ( P < 0.01). Compared with the model group, protein activities of NF-κB/P65 and NF-κB/P50 significantly decreased in high and middle dose JJR groups and the tripterygium control group ( P < 0.05, P < 0.01). All indices mentioned above were higher in the low dose JJR group than in the tripterygium control group ( P < 0.05, P < 0.01). Conclusion JJR could lower the expression of serum IL-17 in CIA model rats, and inhibit protein activities of NF-κB/P65 and NF-κB/P50.
11. Effects of Longzuan Tongbi Recip e on Fas/FasL System in Serum and Synovium of Collagen-induce d Arthritis Rats
Chinese Journal of Integrated Traditional and Western Medicine,Part 1: Mechanism Study,Vol 36,No. 11
Objective To observe the effect of Longzuan I ongbi Recipe (LTR) on Fas/FasL systems in serum and synovium of collagen-induced arthritis (CIA) rats. Methods Ten rats were randomly selected from 60 male Wistar rats as a normal control group. CIA model was prepared by injecting type II bovine collagen and incomplete Freund’s adjuvant mixture in the rest 50 rats. After modeling rats were divided into the model group, the methotrexate (MTX) group, high- , middle- , and low-dose LTR groups, with 10 in each group. Normal saline was administered to rats in the model group by gastrogavage. MTX solution (0.27 mg/100 g) was administered to rats in the MTX group by gavage, once per week for four successive weeks. LTR (4.3 2, 2.16, 1.08 g/mL) was administered to rats in the three LTR groups by gavage, twice per day for 30 successive days. Morphological changes of synovium were observed by HE staining. Expression levels of Fas/FasL in rat serum and synovium were quantitatively detected by ELISA. Results Normal synovium cells could be seen in the normal group. But they were obviously proliferated, fat cells in the lower synovium were reduced or deformed, fibroblasts were increased in the model group, accompanied with infiltration of lymphocytes and monocytes. All these changes were more obviously alleviated in the MTX group, and the three LTR groups. Compared with the normal control group, Fas expression level increased in rat serum and synovium, serum FasL expression level decreased in the model group ( P < 0.05, P < 0.01). Compared with the model group, Fas expression levels were decreased in rat serum and synovium in the MTX group, high- and middle-dose LTR groups. Fas expression levels in rat serum were increased in the MTX group and three LTR groups; Fas expression levels in synovium were increased in the MTX group, high- and middle-dose LTR groups ( P < 0.05, P < 0.01). Compared with the MTX group, Fas expression level in serum of the low-dose LTR group, and Fas expression levels in synovium of low- and middle-dose LTR groups were elevated; Fas expression level in serum and synovium of the high-dose LTR group was reduced; FasL expression levels in serum and synovium of low- and middle-dose LTR groups were reduced; FasL expression levels in serum and synovium were increased in the high-dose LTR group ( P < 0.0 5, P < 0.01). Conclusion LTR could control and treat rheumatoid arthritis, and its mechanism might lie in regulating Fas/FasL systems mediated cell apoptosis, and relieving pathological reaction of rheumatoid arthritis.
12. Effects of Wenhua Juanbi Recipe on the Expressions of DNA Methyltransferases in Peripheral Blood Mononuclear Cells of Collagen-Inducing Arthritis Rats
Chinese Journal of Integrated Traditional and Western Medicine,Part 1: Mechanism Study,Vol 36,No. 12
Objective To observe the effect of Wenhua Juanbi Recipe (WJR) on the expressions of DNA methyltransferases (DNMTs) in peripheral blood mononuclear cells (PBMCs) of collagen-inducing arthritis (CIA), and to study its mechanism for treating CIA. Methods Totally 90 Wistar rats were randomly divided into the model group ( n = 80) and the normal control group ( n = 10). Rats of the model group were injected with type Ⅱ collagen of bovine (BCⅡ) emulsion from the tail to establish CIA model. The successfully modeled 50 CIA rats were randomly divided into five groups, i. e., the model group, the methotrexate (MTX) group, the low dose WJR group, the middle dose WJR group, the high dose WJR group, 10 in each group. The rats in the model group were administered with normal saline by gastrogavage, once per day. The rats in low, middle, and high dose WJR groups were administered with WJR by gastrogavage at the daily dose of 22.9, 45.8, and 68.7 g/kg, respectively (once per day). The rats in the MTX group were administered with MTX suspension (0.78 mg/kg) by gastrogavage, once per week for 30 successive days. The paw swelling was evaluated using volume method (draining volume). PBMCs were extracted from each group after intervention. The mRNA expression levels of DNMTs (DNMT1, DNMT3a, and DNMT3b) were detected by real-time quantitative PCR. Results Compared with the normal group, the paws were obviously swollen in the model group ( P < 0.01). Compared with the model group, swollen paws were obviously alleviated in low, middle, and high dose WJR groups, and the MTX group ( P < 0.01). Compared with before treatment in the same group, swollen paws were obviously alleviated in low, middle, and high dose WJR groups, and the MTX group ( P < 0.01). Compared with the normal group, the expression levels of DNMT1, DNMT3a, and DNMT3b in PBMCs were obviously lowered in the model group ( P < 0.01). Compared with the model group, the expression levels of DNMT1, DNMT3a, DNMT3b in PBMCs were obviously elevated in low, middle, and high dose WJR groups, and the MTX group (all P < 0.01). There was no significant difference in the expression levels of DNMT1, DNMT3a, or DNMT3b in PBMCs among low, middle, and high dose WJR groups ( P > 0.05). Conclusions The expression levels of DNMTs in PBMCs of CIA rats decreased. WJR up-regulated the expression level of DNMTs in PBMCs of CIA rats in an no obvious dose dependent way. One of WJR’s mechanisms for treating CIA might be up-regulating the expression levels of DNMTs, and adjusting the state of DNA methylation.
13. Experimental Study on Therapeutic Effect of Zusanli Acupoint Injection of Bee Venom in Adjuvant-Induced Arthritis Rats
Liaoning Journal of Traditional Chinese Medicine,Part 1: Mechanism Study,Vol 44,No. 13
Objective: To investigate specific mechanism of bee therapy on rheumatoid arthritis through the experimental study on the therapeutic effect of Zusanli (ST36) acupoint injection of bee venom in adjuvant-induced arthritis rats. Methods: Thirty rats were randomly divided into 3 groups as follows: bee venom treatment group, model group, blank group. Blank control group rats were injected saline 0.1 mL. Other two groups were made adjuvant-induced arthritis by CFA 0.1 mL. Each group included 10 rats. The bee venom treatment group rats were daily injected bee venom 0.3 mg in acupoint Zusanli (ST36). Record the thickness and heat pain threshold of the rats’ paws. Detect the level of IL-1β in the peripheral blood by ELISA. Results: Bee venom group rats’ thickness and heat pain threshold were lower than those of model group and blank group. The detection of IL-1β level in each group (from high to low) was model group, bee venom group, blank group. Conclusion: Bee venom can mitigate AA rats by reducing swelling, improving the heat pain threshold and decreasing the IL-1β level. Zusanli (ST36) acupoint injection of bee venom can be effective in treatment of rheumatoid arthritis.
China Journal of Chinese Materia Medica,Part 1: Mechanism Study,Vol 42,No. 14
To explore the prevention and protection effect of Diosocorea nipponica (DNM) on acute gouty arthritis (AGA) rats based on liver metabonomics, and find potential biomarkers and related pathways. AGA model rats were induced by monosodium urate crystal suspension. The UPLC-TOF-MS coupled with pattern recognition technique was employed to find out the potential biomarkers and related metabolic pathways. Eleven common potential biomarkers were identified. Among the potential intervention targets in normal rats given by DNM, 4 biomarkers were up-regulated, and the other 4 targets were down regulated. Among the potential intervention targets in AGA rats given by DNM, 5 metabolites were up-regulated by MSU and 5 metabolites were down regulated. The abnormal expression levels of adenosine monophosphate, 5-methyltetrahydrofolic acid, oxidized glutathione, hypoxanthine, docosahexaenoic acid, glutathione, uridine diphosphate glucose and inosine could be corrected by DNM extract. Three pathways were founded with the greatest correlation, including purine metabolism, starch and sucrose metabolism and glutathione metabolism. Therefore, it could be inferred that D. nipponica had the effect for anti-acute gouty arthritis by intervening endogenous metabolites from the liver under physiological condition and acute gouty arthritis condition.
15. Influence of Bushen Qutan Formula on Leptin, Ob-Rb, TNFα, INFγ, PGE 2, MMP-9, MMP-13 and MMP-1 mRNA Expression Levels in Obese Rat Model with OA due to Kidney-deficiency
Chinese Archives of Traditional Chinese Medicine,Part 1: Mechanism Study,Vol 35,No. 15
Objective: To study the effects of Bushen Qutan Formula on leptin, Ob-Rb, TNFα, INFγ, PGE 2, MMP-9, MMP-13 and MMP-1 mRNA expression levels in obese rats with osteoarthritis (OA) due to kidney-deficiency by PCR. Method: The rats were dividied into normal group, model group, low-dose, middle-dose and high-dose Bushen Qutan Formula groups, and glucosamine sulfate capsule suspension group. Rats in the normal group and model group were administered with normal saline by gavage each day and those in the various treatment groups were administered with the corresponding doses for seven continuous days by gavage. Rats were killed after the last drug administration. In the bacteria-free environment, we decalcified the cartilage and studied the levels of leptin, Ob-Rb, TNFα, INFγ, PGE 2, MMP-9, MMP-13 and MMP-1 mRNA expression in rats by PCR after obtaining of the cartilage. Result: Compared with the normal group, there was no statistical significances in the mRNA expression levels of Ob-Rb, TNFα, INFγ, PGE 2, MMP-9 and MMP-1 after the intervention of Bushen Qutan Formula, while there were significantly statistical significances in the mRNA expression levels of leptin and MMP-13. The mRNA expression levels of leptin and MMP-13 in the model group were increased ( P < 0.01). Compared with the model group, the mRNA expression levels of leptin and MMP-13 in low-dose, middle-dose, high-dose Bushen Qutan Formula groups and glucosamine sulfate capsule suspension group were decreased ( P < 0.05), especially in the high-dose Bushen Qutan Formula group. Conclusion: The obese rat model with OA due to kidney-deficiency was stable, which would provide the new method for modeling. Bushen Qutan Formula could decrease the mRNA expression levels of leptin and MMP-13 in obese rats with OA due to kidney-deficiency, which would provide the theoretical basis for clinical study of Bushen Qutan Formula.
The Chinese Journal of Clinical Pharmacology,Part 1: Mechanism Study,Vol 33,No. 16
Objective To observe the inhibitory effect of Curcumin analogue FM0807 on pannus formation and its possible mechanism. Methods A total of 40 male SD rats were used to establish the adjuvant-induced arthritis rat model, and then randomly divided into model group, control group, test A group, test B group and test C group, 8 rats in each group. Another 8 healthy SD rats were used as blank control group. Then, 7 days after modeling, test A group, test B group and test C group were given 25, 50, 100 mg·kg ?1 FM0807 through gavage once a day; control group was given 0.2 mg·kg ?1 dexamethasone intraperitoneal injection, once a day; model group and the blank control group were given 0.9% NaCl gavage, once a day. The rats in the six groups were treated for 21 days. The content of interleukin-6 (IL-6) and tumor necrosis factor- α (TNF- α) in serum was observed by using enzyme-linked immunosorbent assay (ELISA) and c-Jun N-terminal kinase (JNK) protein expression was detected by Western blotting. Results The content of TNF- α and IL-6 in serum of test A group, test B group, test C group, control group, model group and blank control group was (106.10 ± 7.15), (88.12 ± 12.75), (67.18 ± 11.95), (40.96 ± 9.40), (123.76 ± 5.05), (18.02 ± 10.97) pg·mL ?1 and (1.61 ± 0.14), (1.48 ± 0.15), (0.73 ± 0.16), (0.41 ± 0.12), (1.88 ± 0.09), (0.27 ± 0.10) pg·m L ?1, respectively. JNK protein expression was (7.00 ± 1.22), (3.40 ± 0.54), (2.60 ± 0.55), (1.40 ± 0.55), (7.20 ± 1.30), and (1.20 ± 0.27), respectively. The difference between test C group and model group was statistically significant (all Ps < 0.01). Conclusion Curcumin analogue FM0807 may inhibit the formation of pannus and relieve the symptom of rheumatoid arthritis by inhibiting JNK pathway, decreasing the release of inflammatory mediators, and reducing inflammatory cell stimulation.
17. Paeoniflorin-6'-O-benzene sulfonate influences expression of CXCL12/CXCR4 in spleen of adjuvant-induced arthritis
Chinese Pharmacological Bulletin,Part 1: Mechanism Study,Vol 34,No. 17
Aim To observe the expression of CXCL12 and its receptor CXCR4 in spleen of rats with adjuvant induced arthritis (AA) and the effects of paeoniflorin-6'-O-benzene sulfonate (CP-25). Methods AA rats were induced using complete Freund’s adjuvant and were randomly divided into normal group, AA group, CP-25 group (50 mg·kg ?1) and methotrexate group (MTX, 0.5 mg·kg ?1), which were treated from d 14 to d 28. HE staining was used to assess the pathological changes of spleen. The expression of CXCL12 and CXCR4 in spleen was detected by ELISA, immunohistochemistry and Western blot. Results CP-25 (50 mg·kg ?1) alleviated the pathological changes of spleen and decreased the expression of CXCL12 and CXCR4 in spleen of AA rats. The pathological changes of spleen and the expression of CXCL12/CXCR4 in spleen revealed a positive correlation. Conclusions Increased expression of CXCL12 and its receptor CX-CR4 may be associated with the pathological changes of spleen in AA rats, which plays an important role in the pathogenesis of RA. CP-25 has obvious therapeutic effect on AA rats and its mechanism may be related to the inhibition of the expression of CXCL12 and CXCR4 in the spleen.
18. A comparative study on the mechanisms of two classical herbal formulae for rheumatoid arthritis applying cold and heat patterns based on target network
Acta Pharmaceutica Sinica,Part 1: Mechanism Study,Vol 53,No. 18
Wutou Decoction (WTD) and Baihu-Guizhi Decoction (BHGZD) as described in the Synopsis of the Golden Chamber have been used extensively for the treatment of rheumatoid arthritis (RA) with apparent therapeutic efficacy. However, characteristics of pharmacological effects and their underlying molecular mechanisms have not been fully elucidated due to a lack of appropriate scientific methodology. In the current study, we performed an integrative approach applying gene expression profiling and network analysis to examine the therapeutic effects and molecular mechanisms of WTD and BHGZD based on adjuvant-induced arthritis (AIA) animal model. Results demonstrated that both WTD and BHGZD could relieve the severity of arthritis in AIA rats, while the significant differences were observed in the changes of the withdrawal response scores and latency time of AIA rats treated with WTD and BHGZD. Our network pharmacology-based investigation demonstrated that the major candidate targets of WTD and BHGZD were significantly associated with several inflammation-immune regulatory pathways, such as Toll-like receptor signaling pathway, T cell receptor signaling pathway, cytokine-cytokine receptor interaction, chemokine signaling pathway, B cell receptor signaling pathway, antigen processing and presentation, Fc epsilon RI signaling pathway, natural killer cell mediated cytotoxicity, as well as leukocyte transendothelial migration. In particular, the major candidate targets of WTD were also involved in the regulation of hormone and energy metabolism, which might be imbalanced during RA progression. In conclusion, the current study revealed differences and similarities regarding the effects and network regulatory mechanisms of WTD and BHGZD. These findings may present a scientific basis for elucidation of mechanisms by which WTD and BHGZD alleviates RA.
19. Identification of biomarkers to response of Tripterygium Glycosides Tablets acting on rheumatoid arthritis by integrating transcriptional data mining and biomolecular network analysis
China Journal of Chinese Materia Medica,Part 1: Mechanism Study,Vol 44,No. 19
Growing clinical evidence shows that a partial rheumatoid arthritis (RA) patient treated with Tripterygium Glycosides Tablets (TGTs) may fail to achieve clinical improvement. It is of great clinical significance to predict the therapeutic effect of TGTs in RA. Therefore, the aim of the current study was to identify potential biomarkers for TGT treatment in RA. Affymetrix EG1.0 arrays were applied to detect gene expression in peripheral blood mononuclear cells obtained from six RA patients (three responders and three non-responders) treated with TGTs. By integrating differential expression data analysis and biomolecular network analysis, 360 mRNAs (185 up-regulated and 175 down-regulated) and 24 miRNAs (seven up-regulated and 17 down-regulated) which were differentially expressed between TGT responder and non-responder groups were identified. A total of 206 candidate target genes for the differentially expressed miRNAs were obtained based on miRanada and Target Scan databases, and then the miRNA target gene co-expression network and miRNA-mediated gene signal transduction network were constructed. Following the network analyses, three candidate miRNAs biomarkers (hsa-miR-4720-5p, hsa-miR-374b-5p, hsa-miR-185-3p) were identified as candidate biomarkers predicting individual response to TGT. Partial least-squares (PLS) was applied to construct a model for predicting response to TGT based on the expression levels of the candidate gene biomarkers in RA patients. The five-fold cross-validation showed that the prediction accuracy of this PLS-based model efficacy was 100.00%, 100.00%, 100.00%, 66.67% and 66.67% respectively, and all the area under the receiver operating characteristic curve were 1.00, indicating the highly predictive efficiency of this PLS-based model. In conclusion, the integrating transcription data mining and biomolecular network investigation show that hsa-mir-4720-5p, hsa-mir-374b-5p and hsa-mir-185-3p may be candidate biomarkers predicting individual response to TGT. In addition, the PLS model based on the expression levels of these candidate biomarkers may be helpful for the clinical screen of RA patients, which potentially benefit individualized therapy of RA in a daily clinical setting.
20. Pharmacokinetics of Achyranthes bidentata on adjuvant arthritis rats by microdialysis and UHPLC-MS/MS
China Journal of Chinese Materia Medica,Part 1: Mechanism Study,Vol 44,No. 20
To investigate the “drug-guide” effect of Achyranthes bidentata saponins (ABS) and geniposide (GE) in the treatment on adjuvant arthritis (AA) rats. A UHPLC-MS/MS method for the quantitative determination of GE, zingibroside R 1, ginsenoside Ro and chikusetsu saponin Ⅳa in rat blood and joint dialysate was established. After single or combined administration with ABS and GE to AA rat model, a microdialysis sampling method for rat joint cavity and jugular vein blood vessels was established to collect microdialysis samples. Waters Acquity HSS C 18 column was used to separate the above four components, with acetonitrile-0.1% formic acid water as mobile phase for gradient elution. ESI source was adopted for mass spectra in a negative ion scanning mode. Multiple reaction monitoring (MRM) mode was applied to detect the above four components. The methodological results showed that GE, zingibroside R 1, ginsenoside Ro and chikusetsu saponin Ⅳa demonstrated a good linear relationship within the concentration ranges of 2–4 000, 16–4 096, 14–3 584, 23–5 888 μg·L ?1 respectively. The precision, accuracy, stability and matrix effect of these four ingredients reached the requirements of quantitative analysis of biological samples. The pharmacokinetic results demonstrated that the combined administration of ABS and GE (60 mg·kg ?1 + 60 mg·kg ?1) could increase the degree of GE in joint cavity distribution, and the AUC joint/AUC plasm were twice of that of single administration of GE (60 mg·kg ?1), which indicated that ABS might played a vital role in GE’s distribution to joint cavity. Moreover, there was no significant difference between the distribution trend of total three ABS and GE in rats. The pharmacodynamics results showed that the combined administration of ABS and GE had stronger effects on paw swelling, arthritis index and synovial pathomorphology of AA rats than single administration of GE, which suggested that ABS might improve GE’s anti-inflammatory effect in AA rats. Based on the above results, ABS has a targeting effect in increasing GE’s concentration in joint cavity, with a synergy in efficacy.
21. Effect on Bushen Huoxue Formula on Construction and Distribution of Lymphatic Vessels around Knee Joint in Osteoarthritic Mice
Chinese Archives of Traditional Chinese Medicine,Part 1: Mechanism Study,Vol 36,No. 21
Objective: To investigate the regulating effect of Bushen Huoxue Formula (BSHXF) on the structure and distribution of local lymphatic vessels in osteoarthritis (OA) mice, and further explore its therapeutic effect on joint degeneration. Methods: Thirty male C57BL/6 mice of clean grade were randomly divided into three groups, with 10 mice in each group. The mice in the sham operation group only received the opening of the skin and joint capsule on the medial side of the right knee, with no structure damaged. The mice in the other groups were modeled by cutting off the medial collateral ligament and the anterior medial meniscus. The ones in the BSHXF group were administrated with BSHXF at a dose of 0.2 mL per mouse by gavage since the second day after modeling, while those in the sham operation group and the model group received the same dose of 0.9% sodium chloride solution, once a day for 12 weeks. Mice were sacrificed in the 4th and 12th weeks to harvest the right knee joint. Safranin-O staining was conducted to observe the morphology and structure of joint tissues. The immunofluorescence double staining and whole-slide imaging system (WSIS) were used to detect the local lymphatic vessel structure and distribution in mice. Results: In the 12th week after operation, the knee joint of the model group showed typical OA changes such as cartilage fissures, joint free bodies, and articular cartilage loss. The OARSI score was significantly increased ( P < 0.01). The lymphatic capillaries and total lymphatic vessels were increased ( P < 0.01 and P < 0.05), while the mature lymphatic vessels were decreased ( P < 0.05). Twelve weeks after surgery, the mature lymphatic vessels and total lymphatic vessels of mice in the BSHXF group were increased significantly ( P < 0.01), and the OARSI score was decreased significantly (compared with that in the model group, P < 0.05). Conclusion: BSHXF may delay joint degeneration by promoting lymphatic maturation and increasing its circulation function.
Chinese Archives of Traditional Chinese Medicine,Part 1: Mechanism Study,Vol 37,No. 22
Objective: To investigate the effect of ursolic acid (UA) on the imbalance of peripheral T lymphocyte T helper 17 (Th17)/regulatory T cell (Treg) in peripheral blood lymphocytes of rats with collagen-induced arthritis (CIA). Methods: Female Sprague-Dawley (SD) rats were selected and duplicated into CIA model. After successful modeling, CIA rats were randomly divided into a CIA model group, a low-dose UA group, a high-dose UA group, a MTX group and a UA + MTX group for corresponding treatments. On the 21 st day of primary immunization, the peripheral blood lymphocytes from each group were collected and stained. Peripheral blood Th17/Treg ratio was measured by flow cytometry. CD + 4 T cells were isolated for detecting the mRNA expression levels of retinoid acid-relatedorphan receptor gamma t (RORγt), fork head box P3 (Foxp3), and suppressor of cytokine signaling 3 (SOCS3) by RT-PCR. Results: (1) Compared with the normal control group, the proportion of Th17 cells in the peripheral blood of the CIA model group was significantly increased. Compared with the CIA model group, the proportions of Th17 cells in the treatment groups (high-dose UA group, MTX group, and UA + MTX group) were significantly reduced and the differences were statistically significant ( P < 0.01). (2) Compared with the normal control group, the proportion of Treg cells in the peripheral blood of the CIA model group was significantly down-regulated. Compared with the CIA model group, the proportions of Treg cells in the treatment groups (high-dose UA group, MTX group, and UA + MTX group) were increased and the differences were statistically significant ( P < 0.05). (3) Compared with the normal control group, the RORγt mRNA expression in the CIA model group was increased, while FoxP3 and SOCS3 mRNA expression levels were decreased. Compared with the model control group, the RORγt mRNA expression in each treatment group was decreased. FoxP3 mRNA expression levels of the high-dose UA group, MTX group, and UA + MTX group were increased and SOCS3 mRNA expression levels of the low-dose UA group, high-dose UA group, and UA + MTX group were increased. Conclusion: UA at the high dose and UA + MTX inhibited the differentiation of CD 4 +T cells into Th17 cells, reduced the proportion of Th17 cells, increased the proportion of Treg cells, and maintained the balance of Th17/Treg cells, which might be the possible mechanisms for UA in alleviating the joint inflammation of experimental CIA model.
23. Comparative study on dose-toxicity-effect of Tripterygium Glycosides Tablets and Leigongteng Tablets on CIA model rats
China Journal of Chinese Materia Medica,Part 1: Mechanism Study,Vol 44,No. 23
The aim of this paper was to compare the properties of Tripterygium Glycosides Tablets (TG) and Tripterygium wilfordii Tablets (TW) from dose–effect–toxicity on type Ⅱ collagen-induced arthritis (CIA) in rats. The SD rats were randomly classified into normal group, model group, TG groups (1 time of equivalent dose of 0.009 g·kg ?1, 4 times of equivalent dose of 0.036 g·kg ?1, 16 times of equivalent dose of 0.144 g·kg ?1), TW groups (1 time of equivalent dose of 0.007 5 mg·kg ?1, 4 times of equivalent dose of 0.030 mg·kg ?1, 16 times of equivalent dose of 0.120 mg·kg ?1). From the first immunization, TG and TW were administered intraperitoneally once a day. After the second immunization, the symptoms such as redness and swelling of joints were observed, and the clinical score and incidence of arthritis were evaluated. HE staining and Masson staining were used to examine the histopathological changes of joints. The expression level of anti-type Ⅱ collagen antibody IgG in serum was detected by ELISA, followed by routine testing of blood components. The concentrations of ALP (alkaline phosphatase), ALT (alanine aminotransferase), AST (aspartate aminotransferase), GGT (gamma-glutamyltransferase), TBiL (total bilirubin), CRE (creatinine) and UREA (urea) in serum were detected by enzymatic assay. The rate of sperm deformity in the epididymis was evaluated under light microscope. The extent of damage to the testis and ovarian tissue was assessed by HE staining. TG and TW attenuated the inflammation, redness, swelling and deformity of joints and reduced the clinical score and incidence of arthritis in CIA rats. Meanwhile, it also exhibited obvious reduction in all pathological features such as joint synovitis, pannus, cartilage erosion and bone destruction. TG and TW reduced the IgG in a dose-dependent manner, and TG was better than TW ( P < 0.05 or P < 0.01). The high doses of TG and TW could significantly increase the organ coefficient of liver and spleen and reduced RBC and HGB in CIA rats ( P < 0.01), and severity leading to death. Gastric mucosal injury and morphological changes of liver and kidney were not observed in CIA rats of TG and TW treatment groups. The 4 and 16 times doses of TG and TW could significantly increase the serum ALT and GGT and decrease the CRE ( P < 0.05 or P < 0.01). TG and TW could increase the sperm deformity rate and damage the testicular seminiferous tubules of CIA male rats. The severity increased with dose and time increasing. The effect of TG (16 times) was more significant than TW (16 times). TG and TW significantly delayed the onset of arthritis and inhibited the paw edema and arthritic score. TG and TW also caused the male reproductive damage; high dose affected the hematopoiesis and maximum dose led to death. TG and TW all depended on a dose–effect–toxicity manner. The anti-arthritis effect of TG was better than that of TW, but the toxicity of TG maximum dose was more obvious. The relevant conclusions of our study will provide experimental references for clinical rational use of drugs, and further clinical studies are needed to confirm our conclusions.
World Chinese Medicine,Part 1: Mechanism Study,Vol 14,No. 24
Objective: To study and analyze the mechanism of icariin on TDP-43 mediated chondrocyte lesions in osteoarthritis (OA), so as to provide a reference for effective treatment of chondrocystic lesions in OA. Methods: TDP43 lentivirus was transfected into human chondrocytes, and intervened by icariin with different concentrations. Fluorescence quantitative PCR was used to detect the expression of TDP-43 gene in different groups of human chondrocytes. Flow cytometry was conducted to detect the apoptosis of human chondrocytes transfected with TDP43 lentivirus before and after the addition of icariin. We compared the expression levels of TDP-43 and collagen type I α 1 in human chondrocytes, the apoptosis of TDP-43 lentivirus transfected human chondrocytes before and after icarrin treatment, as well as the number of chondrocytes after treated with different concentrations of icarrin. Results: TDP-43 gene expression level in human chondrocytes transfected with TDP-43 lentivirus was (16.02 ± 1.45), which was higher than (1.86 ± 0.13) and (1.88 ± 0.12) in the normal human chondrocytes and human chondrocytes transfected with blank lentiviral vector , with statistically significant differences (all P < 0.05). The collagen type I α 1 in human chondrocytes of the three groups were not statistically significant ( P > 0.05). The apoptosis rate of human chondrocytes transfected with TDP43 lentivirus after the addition of different concentrations of icariin was lower than that before icariin treatment, with statistically significant differences (all P < 0.05). The number of chondrocytes incubated with high-concentration icarrin for 3 d, 6 d, and 12 d, respectively, was significantly higher than those in the medium-concentration icarrin treatment group. And the number of chondrocytes incubated with medium-concentration icarrin was significantly higher than that in the low-concentration icarrin treatment group (all P < 0.05). Conclusion: TDP-43 may play a negative regulatory role in chondrocyte lesion of OA, while icariin can improve the above process and further alleviate the TDP-43 mediated chondrocyticchondrocyte lesions in OA.
25. An exploration into the therapeutic effects and molecular mechanisms of paeoniflorin in the treatment of adjuvant-induced arthritis rats by a network pharmacology-based research strategy
Acta Pharmaceutica Sinica,Part 1: Mechanism Study,Vol 54,No. 25
Paeoniflorin (PAE), the major active compound of Chinese herb Radix Paeoniae Alba and Chinese patent drug “Total Glucosides of Paeony Capsules”, is effective for the treatment of rheumatoid arthritis (RA), and it exerts multi-pharmacological activities, such as anti-inflammation and immunoregulation. However, its potential action mechanisms remain unclear. Herein, we predicted the putative targets of Radix Paeoniae Alba and constructed an interaction network of putative targets of Radix Paeoniae Alba and known RA-related genes. A list of key putative targets was identified by calculating their topological features (degree, node betweenness and closeness) in the above pharmacological network. Importantly, pathway enrichment analysis revealed that these key putative targets were significantly enriched in several RA-related pathways, including cartilage damage-related IL1 B-TNF-TLR2-JUN-MMP1-MMP3 signaling pathway. Further molecular docking simulation showed that PAE, the major active compound of Radix Paeoniae Alba, had strong binding affinity with MMP1 and MMP3 proteins. Next, in vivo experiments based on the adjuvant-induced arthritis (AIA) animal models showed that PAE significantly alleviated the disease severity and the syndromes of severe redness or swelling in hind limbs of AIA rats, including decreasing the arthritis score, the diameter of the limbs, and elevating body weight and pain thresholds (all P < 0.05). ELISA assay indicated that PAE obviously suppressed the abnormal up-regulation of serum inflammatory factors including IL-1 β, TNF- α, IL-6, IL-17 and IFN- γ in AIA rats (all P < 0.001). Western blot analysis revealed that PAE simultaneously modulated the abnormal up-regulation of MMP1 and MMP3 proteins in the ankle tissues of AIA rats (all P < 0.001) (all procedures in the current study were performed in accordance with the ethical standards of the Center for Laboratory Animal Care, China Academy of Chinese Medical Sciences). In conclusion, PAE alleviated the cartilage damage and disease severity in the progressive process of RA via regulating the IL1 B-TNFTLR2-JUN-MMP1-MMP3 pathway. This study has provided the theoretical basis for exploring the pharmacological activities of Radix Paeoniae Alba and PA against RA and also provided a methodological reference for studying the action mechanisms of anti-inflammatory active components from Chinese medicines.