China Journal of Chinese Materia Medica, the 1st in the field of TCM, is supervised by China Association for Science and Technology and sponsored by Institute of Chinese Pharmaceutical Association. The journal is China's earliest comprehensive core journal of traditional Chinese medicine, and always maintains the circulation top in the professional areas. The journal publishes the latest research and progress of traditional Chinese medicine and takes a leading position in numbers of articles published, downloads and citations among all journals in this discipline.
Its scope covers new achievements, technologies, methods, experiences and concepts resulting from the research on Chinese materia medica pursuant to Chinese medical and pharmaceutical theories, traditional experiences, and modern science and technology, including medicinal resources and identification, cultivation, processing, preparation, chemistry, pharmacology, theory of Chinese pharmacy and clinical practice, bencaological study.
The journal is included in CA, JST and CSCD.
Honorary Editor-in-Chief Xiao Peigen Editor-in-Chief Wang Yongyan
Associate Editors Zhang Boli, Hu Zhibi, Yao Xinsheng, Li Lianda, Li Dapeng, Yang Baofeng, Zhou Chaofan, Huang Luqi, Chen Shilin, Li He.
Executive Editorial Board Cai Shaoqing, Chen Shilin
Tripterygium wilfordii is a medicinal plant commonly used in the treatment of rheumatoid arthritis, and with pharmacological activities in anti-tumor and obesity treatment. The known active ingredients in
T. wilfordii are mainly terpenoids, but with very low content. Therefore, the analysis of the biosynthesis pathway of terpenoids in
T. wilfordii has become a research hotspot to solve the problem of its resources. Terpene synthase (TPS) is a key enzyme that catalyzes the formation of a wide variety of terpenoid skeletons. In this study, a gene fragment with an ORF of 1 785 bp was cloned from
T. wilfordii. Bioinformatics analysis was performed using NCBI’s BLASTP, ProtParam and Interpro online tools and MEGA 6.0 software. The response of this gene to methyl jasmonate was also detected by real-time fluorescent quantitative PCR, and its catalytic function was verified by prokaryotic expression and in vitro enzymatic assay. Bioinformatics analysis indicated that the amino acid sequence encoded by this gene had both N-terminal domain and C-terminal domain of TPS, as well as the DDxxD conserved domain of the class I of TPS family. And TwMTS gathered together with TPS-b subfamily in the phylogenetic tree constructed with known homologous TPSs. The results of RT-PCR showed that 50 μmol·L
−1 MeJA 12 h could increase the expression of TwMTS to 735 times in the control group at 12 h, and 1 644 times at 24 h. In addition, in vitro enzymatic reaction results showed that TwMTS can catalyze the production of
β-citronellol with GPP as substrate, indicating that TwMTS was a monoterpene synthase. The above results provided a new element for the synthetic biology database of
T. wilfordii terpenoids, and laid the foundation for future biosynthesis research.
The aim of this paper was to study the influence of triptolide in the immune response pathways of acquired immune deficiency syndrome (AIDS). Target proteins of triptolide and related genes of AIDS were respectively searched in PubChem and Gene databases on line. Molecular networks and canonical pathways comparison analyses were performed by bioinformatics software (IPA). There were 15 targets proteins of triptolide and 258 related genes of AIDS. Close biological relationships of molecules of triptolide and AIDS were established by networks analysis. There were 21 common immune response pathways of triptolide and AIDS, including neuroinflammation signaling pathway, Th1 and Th2 activation pathway, and role of pattern recognition receptors in recognition of bacteria and viruses. Triptolide stimulated immune response pathways mainly by IFNγ, JAK2, NOD1, PTGS2, and RORC. IFNγ is the focus node of triptolide and AIDS, and directly or indirectly regulates the genes of AIDS. Triptolide may act against AIDS by regulating IFNγ in immune response pathways.
The aim of this paper was to observe the concentration, time and mechanism of autophagy induced by triptolide (TP) in ovarian granulosa cells (OGCs). CCK-8 method was used to compare the inhibitory effects of TP at different concentrations on primary cultured rat OGCs and IC
50 was calculated. The effects of TP at different concentrations and time points on the expression of OGCs autophagy factor protein and the cascade of PI3K/AKT/mTOR pathway were detected by Western blot. The effects of TP, autophagy inducer (brefeldin A) and PI3K/mTOR inhibitor (NVP-BEZ235) on the expression of PI3K/AKT/mTOR cascade and autophagy related factor protein were detected by Western blot. The results showed that the IC
50 of different concentrations of TP on OGCs of rat ovary was 14.65 μmol·L
−1, and the minimum inhibitory concentration of TP was 0.1 μmol·L
−1 (100 nmol·L
−1). Compared with the control group, the expression levels of beclin1 and LC3 Ⅱ in each group were significantly higher than those in the control group (
P < 0.05 or
P < 0.01). After 12 h of treatment with TP, brefeldin A and NVP-BEZ235, respectively, compared with the control group, TP could significantly promote the expression level of downstream autophagy effect or molecule beclin1, LC3 Ⅱ and inhibit the expression level of LC3 Ⅰ and p62 protein (
P < 0.05 or
P < 0.01). Moreover, the expression of beclin1 and LC3 Ⅱ/LC3 Ⅰin TP group was higher than that in brefeldin A group (
P < 0.05 or
P < 0.01), and the expression of p62 in TP group was lower than that in brefeldin A group (
P < 0.05 or
P < 0.01). At the same time, TP could significantly inhibit the expression of p-PI3K, p-AKT, p-mTOR protein, and the inhibitory effect of TP was better than that of NVP-BEZ235 group. This study suggests that 100 nmol·L
−1 TP could induce OGCs autophagy successfully in cultured rat ovary for 12 h; TP may induce OGCs autophagy by inhibiting PI3k/Akt/mTOR signaling pathway.
The mass spectrometry-based metabolomics method was used to systematically investigate the formation of celastrol metabolites, and the effect of celastrol on endogenous metabolites. The mice plasma, urine and feces samples were collected after oral administration of celastrol. Ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was applied to analyze the exogenous metabolites of celastrol and its altered endogenous metabolites. Mass defect filter was adopted to screen for the exogenous metabolites of celastrol. Multivariate statistical analysis was used to identify the endogenous metabolites affected by celastrol. Celastrol and its eight metabolites were detected in urine and feces of mice, and five metabolites of them were reported for the first time. The hydroxylated metabolites were observed in the metabolism of both human liver microsomes and mouse liver microsomes. Further recombinant enzyme experiments revealed CYP3A4 was the major metabolic enzyme involved in the formation of hydroxylated metabolites. Urinary metabolomics revealed that celastrol can affect the excretion of intestinal bacteria-related endogenous metabolites, including hippuric acid, phenylacetylglycine, 5-hydroxyindoleacetic acid, urocanic acid, cinnamoylglycine, phenylproplonylglycine and xanthurenic acid. These results are helpful to elucidate the metabolism and disposition of celastrol in vivo, and its mechanism of action.
Tripterygium wilfordii multiglycoside (GTW), an extract derived from
T. wilfordii, has been used for rheumatoid arthritis and other immune diseases in China. However its potential hepatotoxicity has not been investigated completely. Firstly, the content of triptolid (TP) in GTW was 0.008% confirmed by a LC method. Then after oral administration of GTW (100, 150 mg·kg
−1) and TP (12μg·kg
−1) in female Wistar rats for 24 h, it was found that 150 mg·kg
−1 GTW showed more serious acute liver injury than 12 μg·kg
−1 TP, with the significantly increased lever of serum ALT, AST, TBA, TBiL, TG and bile duct hyperplasia even hepatocyte apoptosis. The expression of mRNA and proteins of liver bile acid transporters such as BSEP, MRP2, NTCP and OATP were down-regulated significantly by GTW to inhibit bile acid excretion and absorption, resulting in cholestatic liver injury. Moreover, GTW was considered to be involved in hepatic oxidative stress injury, although it down-regulated SOD1 and GPX-1 mRNA expression without significant difference in MDA and GSH levels. In vitro, we found that TP was the main toxic component in GTW, which could inhibit cell viability up to 80% in HepG2 and LO2 cells at the dose of 0.1 μmol·L
−1. Next a LC-MS/MS method was used to detect the concentration of triptolid in plasma from rats, interestingly, we found that the content of TP in GTW was always higher than in the same amount of TP, suggesting the other components in GTW may affect the TP metabolism. Finally, we screened the 2.9 (p-glycoprotein) in Caco-2 cells treated with components except TP extracted from GTW, finding that wilforgine, wilforine and wilfordine was the substrate of p-glycoprotein. Thus,we speculated that wilforgine, wilforine and wilfordine may competitively inhibit the excretion of TP to bile through p-glycoprotein, leading to the enhanced hepatotoxity caused by GTW than the same amount of TP.
Terpenoids are main bioactive components in
Tripterygium wilfordii, but the contents of some terpenoids are relatively low. In order to provide scientific evidence for the regulation of terpenoids in
T. wilfordii, this research explored the effect of GR24 on accumulations of four diterpenoids (triptolide, tripterifordin, triptophenolide, and triptinin B) in
T.wilfordii suspension cells by biological technology and UPLC-QQQ-MS/MS. The results indicated that 100 μmol·L
−1 GR24 inhibited the accumulations of triptolide, tripterifordin, triptophenolide, and triptinin B to different degrees. Compared with the control group, the contents of four diterpenoids (in the induced group) were down to 96.59%, 63.80%, 61.02% and 33.59% in 240 h, respectively. Among them, the accumulation of triptinin B was significantly inhibited. In addition, the key time point of inhibitory effect was 120 h after induction with GR24 in some diterpenoids. This is the first systematic study focusing on the effect of GR24 on the accumulation of diterpenoids in
T.wilfordii suspension cells. The dynamic accumulation regularity of four diterpenoids after induced by GR24 was summarized, which laid a foundation for further study on the chemical response mechanism of terpenoids to GR24.
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.
To investigate the effect of triptolide on cognitive dysfunction in vascular dementia (VD) rats and on SIRT1/NF-κB pathway. The fifty healthy male Sprague-Dawley (SD) rats were randomly divided into five groups: Sham operation group (Sham group), VD model group (2 VO group), intraperitoneal injection of triptolide group (TR group), intraperitoneal injection triptolide + intracerebroventricular administration of EX527 group (T + E group), and intracerebroventricular administration of EX527 group (EX527 group). After four weeks of modeling, Morris water maze test and object recognition test were carried out to evaluate the learning and memory abilities of rats. The morphological changes in hippocampus in each group were observed. The activities of SOD and MDA in hippocampus were detected by chemical colorimetry. IL-6 and TNF-α levels were detected by ELISA. Western blot assay was conducted to detect the expression levels of SIRT1, NF-κB, IκBα and caspase 3 in hippocampus. The results showed that compared with the Sham group, the learning and memory abilities of the VD model rats were reduced. The SOD activity in the hippocampus was decreased, while the MDA activity and IL-6 level were increased. The neuronal degeneration changed significantly. The expression levels of SIRT1 and IκBα were decreased, while the expression levels of caspase 3 and NF-κB were significantly increased. After intervention by triptolide, the oxidative stress and the degenerative changes in hippocampus were significantly declined. The protein expression levels of SIRT1 and IκBα were increased, while those of caspase 3 and NF-κB were significantly decreased. After intervention by triptolide and EX527, the expression of SIRT1 was decreased, and the levels of oxidative stress and neuronal degeneration in the hippocampus were aggravated. The learning and memory abilities were reduced. The results showed that triptolide could improve cognitive impairment in VD rats, which might be related to SIRT1/NF-κB signaling pathway.
To further investigate the metabolism of Radix Tripterygii Wilfordii and Radix Paeoniae Alba micro-emulsion gel in vivo, we established the LC-MS/MS method for the determination of triptolide and paeoniflorin in Radix Tripterygii Wilfordii and Radix Paeoniae Alba micro-emulsion gel. The extracorporeal recovery rates of blood probe were measured by the incremental method and decremental method. Meanwhile, the microdialysis (MD) methods of tripterine and paeoniflorin in the skin and blood were established, and the pharmacokinetics of Radix Tripterygii Wilfordii and Radix Paeoniae Alba micro-emulsion gel in skin and blood were studied by MD combined with LC-MS/MS quantitative analysis. The results showed that the established method for the determination of triptolide and paeoniflorin in Radix Tripterygii Wilfordii and Radix Paeoniae Alba micro-emulsion gel exhibited good linearity within the required range. The specificity, recovery rate, and degree of precision for the chromatography all conformed to the research requirements of MD samples. The freeze-thaw stability and the residual effect all conformed to the methodological criteria of bioanalysis. The probe recovery rates measured by the incremental method and decremental method were almost consistent with those in the extracorporeal recovery rate test. The recovery rates of paeoniflorin in skin and blood MD were (30.60 ± 1.09)% and (28.01 ± 1.75)%, respectively. And the recovery rates of triptolide in skin and blood MD were (26.79 ± 2.78)% and (25.39 ± 1.86)%, respectively. The intraday recovery rates of probes were stable within 11 h. The results of the pharmacokinetic study showed that the
Cmax values of triptolide in skin and blood were (148.03 ± 41.51) and (76.77 ± 15.27) μg·L
−1, respectively. And the
Tmax values were (2.33 ± 0.29) and (3.00 ± 0) h, respectively. The AUC
0–11 h values were (2 814.05 ± 1 070.37) and (1 580.63 ± 208.27) μg·h·L
−1, respectively. The MRT
0–11 h values were (4.20 ± 0.33) and (4.54 ± 0.34) h, respectively. The
T1/2 values were (4.61 ± 4.11) and (1.07 ± 0.13) h, respectively. The
Cmax values of paeoniflorin in skin and blood were (991.88 ± 152.22) and (407.02 ± 120.06) μg·L
−1, respectively. The
Tmax values were (2.00 ± 0) h and (2.83 ± 0.29) h, respectively. The AUC
0–11h values were (18 430.27 ± 3 289.35) and (6 338.59 ± 1 659.32) μg·h·L
−1, respectively. The MRT
0–11 h values were (4.29 ± 0.16) and (4.00 ± 0.05) h, respectively. The
T1/2 values were (2.16 ± 0.43) and (1.78 ± 0.48) h, respectively. The results suggested that micro-emulsion gel played a role in forming a skin reservoir through percutaneous penetration. It could not only improve the drug transdermal efficiency but also control the sustained release of the drug, thus forming a long-term effect.
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.
