Suppressive effect of Tripterygium hypoglaucum (Levl.) Hutch extract on rheumatoid arthritis in mice by modulating inflammasome and bile acid metabolism.

Tripterygium hypoglaucum (Levl.) Hutch (THH) has long been used as a remedy for rheumatoid arthritis (RA) in China. However, it is unclear whether the anti-RA mechanism of THH is associated with inflammasome or gut-joint axis. In this study, we aimed to explore the critical role of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) and bile acid (BA) in the anti-RA mechanism. Complete Freund's adjuvant (CFA)-injected mice were treated with THH extract (250 mg/kg/d) for 35 days, and joint swelling and disease scores were measured. After THH treatment, the joint swelling and RA disease score in CFA-treated mice significantly subsided. The increased ratios of lymphocytes, monocytes, and white blood cells were attenuated by THH treatment. Notably, THH treatment blocked the inflammation in both joints and colons by suppressing the NLRP3-mediated inflammasome, as indicated by NLRP3, interleukin 1beta (IL-1ß), and Caspase-1. Meanwhile, THH significantly remodeled the bile acid (BA) profiles in RA mice. Spearman's analysis shed light on the close link between BAs, NLRP3 inflammasome, and RA indicators. However, THH treatment failed to improve inflammasome activation, snoptivis, and joint swelling in RA mice with gut microbiota depletion. In summary, we revealed the pivotal role of BA-mediated gut-joint axis and inflammasome in THH's RA amelioration. In the future, more work should be done to explain the in-depth mechanism between altered BAs and inflammasome.

Tripterygium hypoglaucum (Lévl.) Hutch and Its Main Bioactive Components: Recent Advances in Pharmacological Activity, Pharmacokinetics and Potential Toxicity.

Tripterygium hypoglaucum (Lévl.) Hutch (THH) is believed to play an important role in health care and disease treatment according to traditional Chinese medicine. Moreover, it is also the representative of medicine with both significant efficacy and potential toxicity. This characteristic causes THH hard for embracing and fearing. In order to verify its prospect for clinic, a wide variety of studies were carried out in the most recent years. However, there has not been any review about THH yet. Therefore, this review summarized its characteristic of components, pharmacological effect, pharmacokinetics and toxicity to comprehensively shed light on the potential clinical application. More than 120 secondary metabolites including terpenoids, alkaloids, glycosides, sugars, organic acids, oleanolic acid, polysaccharides and other components were found in THH based on phytochemical research. All these components might be the pharmacological bases for immunosuppression, anti-inflammatory and anti-tumour effect. In addition, recent studies found that THH and its bioactive compounds also demonstrated remarkable effect on obesity, insulin resistance, fertility and infection of virus. The main mechanism seemed to be closely related to regulation the balance of immune, inflammation, apoptosis and so on in various disease. Furthermore, the study of pharmacokinetics revealed quick elimination of the main component triptolide. The feature of celastrol was also investigated by several models. Finally, the side effect of THH was thought to be the key for its limitation in clinical application. A series of reports indicated that multiple organs or systems including liver, kidney and genital system were involved in the toxicity. Its potential serious problem in liver was paid specific attention in recent years. In summary, considering the significant effect and potential toxicity of THH as well as its components, the combined medication to inhibit the toxicity, maintain effect might be a promising method for clinical conversion. Modern advanced technology such as structure optimization might be another way to reach the efficacy and safety. Thus, THH is still a crucial plant which remains for further investigation.

Role of Glutamine-Glutamate/GABA cycle and potential target GLUD2 in alleviation of rheumatoid arthritis by Tripterygium hypoglaucum (levl.) Hutch based on metabolomics and molecular pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Tripterygium hypoglaucum (levl.) Hutch (Celastraceae) (THH), as a traditional Chinese medicine, was clinically exploited to treat rheumatoid arthritis (RA), yet the underlying mechanism for this effect remains largely unclear. AIM OF THE STUDY: This study aimed to examine the beneficial effects of THH extract (THHE) against rheumatoid arthritis and its regulating role in differential metabolic pathways and potential targets. MATERIALS AND METHODS: In the present study, the Lewis rat model with rheumatoid arthritis induced by adjuvant was established and administrated THHE for 14 days. Untargeted/targeted metabolomics analysis were used for determining the changes of differential metabolites, and molecular docking method was further developed to verify predicted targets and investigate the therapeutic mechanism of THH extract on RA. RESULTS: The results showed that THH extract could obviously improve body weight, significantly decrease the joint index and swelling degree of the RA model rats to reduce damage in the joint. Meanwhile, THHE could significantly suppress the releases of IL-1α, IL-1ß and MMP3, but also the expression levels of IL-4 and IL-10 and percentage of Treg cells were significantly improved, a result consistent with inhibitory effects on multiplication of macrophages, inflammatory cell infiltration and fibro genesis in the synovial tissues. Furthermore, 516 differential metabolites were identified by serum metabolic profiles analysis, including vitamin, organic acids and derivatives, lipids and lipid-like molecule, hormone, amino acids and derivatives, and other compounds, which targeted 47 metabolic pathways highly correlated with immunosuppression, such as citrate cycle (TCA cycle), sphingolipid metabolism, urea cycle, arachidonic acid metabolism and amino acid metabolism (such as Glutamine-Glutamate metabolism). Targeted metabolomics was used to verify that L-Glutamate and Glutamine changed significantly after THHE administration for 14 days, and many active ingredients of THHE could be successfully docked with glutamate dehydrogenase 2 (GLUD2). CONCLUSION: This study indicated that the Glutamine-Glutamate/GABA cycle played essential regulation roles in protective effect of THHE on rat RA following adjuvant-induced damage, and GLUD2 as an attractive target also provides great potential for development of therapy agents for rheumatoid arthritis and autoimmune diseases with less unfavorable tolerability profile.

Chemical constituents of Tripterygium hypoglaucum / 中草药

Objective: To study the chemical constitutes from the roots of Tripterygium hypoglaucum. Methods: The chemical constituents from 95% EtOH extract of the roots of T. hypoglaucum at room temperature were isolated and prepared by Silica gel column chromatography, Sephadex LH-20 column chromatography, and semi-pre HPLC after dichloromethane extraction, and their structures were elucidated by a variety of spectral and spectroscopic techniques. Results: Six compounds were isolated and identified as 2-(5’,7’-dimethoxy-2’,2’-dimethyl-2H-benzopyran-6’-)-3-formyl-5,6-dimethoxy-benzofuran (1), 3(R)-4’,5’-dihydroxy- 2’,5,7-trimethoxy-6-(3-methyl-but-2-enyl)-isoflavan (2), 3’-geranyl-5,7,2’,5’-tetrahydroxyisoflavone (3), β-sitosterolpalmitate (4), docosanoic acid 2’,3’-dihydroxypropyl ester (5), and β-sitosterol (6). Conclusion: Compound 1 is a new compound, named as hirtellanine C. Compounds 2-5 are isolated from the plants for the first time.

Chemical constituents from rhizome of Tripterygium hypoglaucum / 中草药

Objective: To study the chemical constituents from the rhizome of Tripterygium hypoglaucum. Methods: The concrete exacted by 95% ethanol from the rhizome of T. hypoglaucum was isolated and purified by chromatography on silica gel column chromatography, gel column chromatography, MPLC, etc. The structures of the chemical constituents were elucidated by means of NMR, MS, etc. Results: Twelve compounds were isolated and identified as 3β-hydroxy-4-hydroxymethyl-abieta-8,11,13-trien-7-one (1), 3β-acetoxy oleanolic acid (2), physcion (3), canophyllal (4), chrysophenol (5), quinone 21 (6), tripterifordin (7), triptobenzene B (8), 3,4-dimethoxyphenol (9), integracin A (10), celastrol (11), and β-sitosterol (12). Conclusion: Compound 1 is a new diterpene named tripterone, compounds 8-10 are isolated from this plant for the first time.

Total alkaloids of Tripterygium hypoglaucum (levl.) Hutch inhibits tumor growth both in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Tripterygium hypoglaucum (levl.) Hutch (Celastraceae) (THH) root is a traditional Chinese medicinal herb commonly used for treating autoimmune diseases and cancer. Alkaloid is one of the most bioactive components of THH extract. To evaluate the in vitro and in vivo antitumor properties of the total alkaloids of THH (THHta). MATERIALS AND METHODS: THHta was extracted in pilot-scale. HCT116 cells were chose to establish human colon cancer xenograft model. The in vitro anti-tumor activity of THHta was tested by Cell malignant transformation test, Soft agar colony formation assay and MTT assay. The in vivo anti-tumor effect of THHta was confirmed by xenograft mouse model. THHta-induced apoptosis was examined by flow cytometry. The levels of apoptosis-related proteins were investigated by Western blot. RESULTS: TPA-induced cell transformation was significantly inhibited by THHta in JB6 Cl41 cells. THHta inhibits the growth of colon cancer cells in vitro in a significant dose-dependent manner. Compared to the control set, i.p. administration of THHta to xenograft mice significantly reduced both tumor weight and volume. Apoptosis induction of THHta was mediated by activation of caspase-3, PARP and inhibiting of Bcl-2, Bcl-xL and XIAP. CONCLUSION: THHta was effective in inhibiting tumor growth both in vitro and in vivo at less toxic concentrations by inducing apoptosis which suggested it could be developed as a potential anticancer agent.

The effect of different regions of Tripterygium Hypoglaucum (Lévl.) Hutch on macrophage inflammatory factor / 国际中医中药杂志

Objective To observe the effect of different regions of tripterygium hypoglaucum (Lévl.) hutch on macrophage inflammatory factor, and to providetheoretical basis and experimental basis for the clinical application of tripterygium hypoglaucum (Lévl.) hutch. Methods 5 batches of tripterygium hypoglaucum (Lévl.) hutch were collected, then the samples turned into alcohol extract by extraction and isolation. The IC50 values of alcohol extracts were measured by MTT in BMDM cell. B MDM cell were induced by the 5 batches of samples with IC50, then IL-6, IL-10, iNOS were detected by Elisa. The efficacy of different regions of tripterygium hypoglaucum (Lévl.) hutch on macrophage inflammatory factor was evaluated by comparison with sulfasalazine. Results The content of IL-6 (4.22 ± 0.38 pg/ml, 4.55 ± 0.44 pg/ml vs.7.92 ± 0.84 pg/ml) and iNOS (0.07 ± 0.04 ng/ml, 0.28 ± 0.10 ng/ml vs. 0.86 ± 0.13 ng/ml) in HuNan and ZheJiang groups were significantly lower than sulfasalazine (P<0.05), and the content of IL-10 (19.34 ± 6.06 pg/ml, 24.34 ± 3.03 pg/ml vs. 9.06 ± 0.40 pg/ml) in Guizhou and Fujian groups were higher than sulfasalazine (P<0.05). Conclusion The anti-inflammatory effect of HuNan and ZheJiang's tripterygium hypoglaucum (Lévl.) hutch treat rheumatoid arthritis is better than sulfasalazine, so theyaregenuine regional drug in the treatment of rheumatoid arthritis. Additional research will analyze associations between tripterygium hypoglaucum (Lévl.) hutch and rheumatoid arthritis.

Research progress on chemical constituents, pharmacological effects, and clinical application of Tripterygium hypoglaucum / 中草药

Tripterygium hypoglaucum (THH) is a folk medicine with important medicinal property, used in the treatment of cataciasis, arthritis, and cancer. The principal chemical constituents of THH are diterpene, triterpene, and sesquiterpene alkaloids, and there are also flavonoid, steroid, tannin, and glucide in it. Diterpene and alkaloids are the major active ingredients. THH has various pharmacological effects such as anti-inflammation, antitumor, immunosuppressive activity, antiviral effect, and so on. It is usually used to treat rheumatiod arthritis, psoriasis, and chronic urticarial. In this paper, we summarize the chemical composition, pharmacological activity, and clinical application of THH, for reference to study and utilize it.

Establishment of hepatotoxic bioassay for quality control of Tripterygium wilfordii / 中草药

Objective: To establish a method suitable to the quality evaluation of the roots in Tripterygium wilfordii based on hepatotoxic potency. Methods: The inhibitory rates of T. wilfordii and its regional substitute-T. hypoglaucum, with normal human hepatocytes (L02 cell line) as carrier, were established by optimizing a series of factors, such as test samples prepared with conditions. Results: Taking p-acetaminophenol as positive control (toxic potency was 400 U/g), The 50% ethanol extract of T. wilfordii was dried under ultrasonic vacuum to obtaine the dry paste. The test solution with crude drug of 3 mg/mL was prepared by medium. The dilution ratio was 1:0.65. The hepatotoxic potencies of 18 batches of T. wilfordii and 5 batches of T. hypoglaucum were 17.78-4131.4 and 209.42-7422.2 U/g, respectively, detected by the reaction parallel line method, and the differences were over 200 and 30 times. There was the significant hepatotoxic potency of T. wilfordii and T. hypoglaucum from the various origins. In addition, the fresh collected samples had the larger hepatotoxic potency than that in the dried samples collected in the market. Conclusion: The preliminarily established hepatotoxic potency bioassay is useful to evaluate the quality of T. wilfordii which directs the correlation with the drug's hepatotoxic adverse effect in clinic. The method can provide the reference for the clinical safety use of T. wilfordii based on the quality control and be as the quantitative method used to screen the toxic components from T. wilfordii.

Study on the acute toxicity of Tripterygium Hypoglaucum (Levl) Hutch solution in mice / 第三军医大学学报

Objective To explore the acute toxicity and LD 50 of Tripterygium Hypoglaucum (Levl) Hutch(THH) solution to provide information for safe clinical application. Methods After oral administration of THH solution in mice, the mortality and the physiological and pathological changes were observed. Results The LD 50 (95% confidence limit) of THH in male and female mice was 79 g/kg(69～89 g/kg) and 100 g/kg (90～112 g/kg), respectively. No marked pathological change of the organs was found. Conclusion According to the standard of grading of acute toxicity, THH solution belongs to the moderate class. Therefore, it is safe in clinical practice and has a wide application.

Role of [Ca~(2+)]i in the apoptosis of Jurkat T lymphoma cells induced by Tripterygium Hypoglaucum (Levl) Hutch alkaloids / 第三军医大学学报

Objective To explore the roles of [Ca 2+ ]i in the apoptosis of Jurkat cells induced by Tripterygium Hypoglaucum (Levl) Hutch alkaloids. Methods After Jurkat cells were stained with Indo 1 AM and PI, changes of [Ca 2+ ]i were assessed by flow cytometry at the single cell level. Results THH alkaloids could induce no changes of [Ca 2+ ] inside cytoplasm. Conclusion THH alkaloids induce the apoptosis of Jurkat cells via other pathways rather than via the [Ca 2+ ] pathway.

Tripterygium Hypoglaucum (Levl) Hutch alkaloids induce DNA strand breaks and DNA fragmentation in Jurkat T lymphoma cells / 第三军医大学学报

Objective To investigate the effects of Tripterygium Hypoglaucum (Levl) Hutch alkaloids on the nuclear DNA strand breaks and DNA fragmantation in Jurkat T lymphoma cells to understand the mechanisms of the apoptosis. Methods After Jurkat cells were induced by the alkaloids, DNA stand breaks were labeled by TUNEL assay and DNA fragmentation was analyzed by DNA content analysis. Flow cytometry was performed to determine these phenomena. Results THH alkaloids could effectively induce DNA stand breaks and DNA fragmentation. Conclusion There are great changes in nuclear DNA in the apoptosis of Jurkat T lymphoma cells induced by THH alkaloids.

Effects of Tripterygium Hypoglaucum (Levl) Hutch alkaloids on cell cycle and time-dependent apoptosis of Jurkat T lymphoma cells / 第三军医大学学报

Objective To study the effects of the cytotoxicity, cell cycle and time dependent apoptosis of Jurkat T lymphoma cells induced by Tripterygium Hypoglaucum (Levl) Hutch (THH) alkaloids so ad to explore the mechanisms of the apoptosis induced by the THH alkaloids. Methods Cell vitality and cell proliferation were measured by Typan blue staining. Cell cycle and the time dependent apoptosis were determined by DNA staining, TUNEL labeling and flow cytometry. Results THH alkaloids could effectively inhibit cell proliferation of Jurkat cells and induce the G 1 arrest and could induce apoptosis in G 2/S phase first and then G 1 phase. Conclusion THH alkaloids can inhibit DNA synthesis, cell proliferation and can also induce apoptosis of Jurkat T lymphoma cells in all phases.

Extract from Tripterygium Hypoglaucum Hutch inhibits neointimal formation of rabbit carotid arteries after balloon angioplasty / 中国介入心脏病学杂志

Objective To study the effects of Tripterygium Hypoglaucum Hutch(THH) and its extract(THW-4) on neointimal formation of rabbit carotid arteries after balloon angioplasty.Methods After injury of the common carotid arteries was set up by balloon angioplasty,the rabbits were divided into 3 groups and were given different treatment protocol including gastrogarage of THH(n=9),associated gastrogavage of THH with local administration of THW-4 into the injured arteries(n=9) and 0.25% DMSO saline local administration as the control(n=9).Neointimal fomation was evaluated on days after the procedure by proliferating all nuclear antigen(PCNA) and Evans Blue staining,and quantitative histomorphometric analysis was performed on 28 days.Results The PCNA-positive index in the neointima at day 14 after the procedure was reduced in the THH gastrogavage group and the THH/THW-4 group compared with the control(17.68%?5.27%,6.60%?2.52% vs 40.37%?7.12%,P