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The Chinese traditional herb Tripterygium wilfordii Hook f (TwHF) has been widely used in the treatment of autoimmune and inflammatory diseases. Over the past few decades, great efforts have been made to explore modern preparations of TwHF with higher efficacy, solubility, and lower toxicity. In this study, we reviewed several examples both of naturally occurring compounds and their derivatives in TwHF, and summarized the preclinical evaluations with regard to autoimmune and inflammatory diseases. All of the candidate compounds described herein have been or are currently in clinical trials. Although some studies encountered problems, the data still provided valuable references for future studies. (5R)-5-hydroxytriptolide (LLDT-8, Leitengshu) is a novel triptolide derivative with potent immunosuppressive and anti-inflammatory activities developed at Shanghai Institute of Materia Medica. Indeed, a Phase I clinical trial for this compound has been completed in rheumatoid arthritis patients. The results will provide the basis for the further exploration of this ancient herb and encourage the research and development of valuable traditional Chinese medicine.

Aim: To examine the inhibitive effects of triptolide on the expression of IL-8, monocyte chemotactic protein (MCP)-1, and matrix metalloproteinases (MMP)-3 in subepithelial myofibroblasts (SEMF) stimulated with IL-1beta., Methods: SEMF cultures were established from normal colons in patients who underwent gut resection for colorectal carcinoma. Chemokine and MMP-3 expressions were determined by ELISA and RT-PCR. The cytosolic amount of phosphorylation of I kappa B-alpha(p-I kappa B-alpha) was determined by Western blotting. The DNA binding capacity of NF-kappa B was evaluated by electrophoretic mobility shift assays., Results: IL-1beta stimulated protein and mRNA expression of IL-8, MCP-1, and MMP-3 in SEMF. Triptolide inhibited these effects of IL-1beta in a dose-dependent manner. Mechanistic studies revealed that triptolide markedly decreased IL-1beta -induced NF-kappa B DNA binding capacity and cytosolic amount of p-I kappa B-alpha. These results showed that triptolide inhibited IL-1beta -induced chemokine and MMP-3 expression in SEMF through the NF-kappa B pathway., Conclusion: Triptolide inhibited IL-1beta -induced chemokine and MMP-3 expression in SEMF by preventing the phosphorylation of I kappa B-alpha.

Aim: To investigate the antiproliferative effect of triptolide on B-NHL cell line Raji cells, to study its effect on lymph node metastasis in patients with non-Hodgkin's lymphoma (NHL) in vitro, and to explore the underlying mechanism regulating SDF-1/CXCR4 axis., Methods: The effects of triptolide on the growth of Raji cells were studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. The effects of triptolide on SDF-1 mRNA expression in lymph node stromal cells from patients with NHL were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The effects of triptolide on CXCR4 expression on lymphoma cells freshly isolated from the lymph nodes of these patients were studied by flow cytometric analysis. Chemotaxis assays were performed to observe the effects of triptolide on migration of primary lymphoma cells towards recombinant human SDF-1 alpha (rhSDF-1 alpha) or cultured lymph node stromal cells in vitro., Results: Triptolide inhibited the proliferation of B-NHL cell line Raji cells in a dose- and time-dependent manner with a 24-h IC50 value of 43.06 nmol/L and a 36-h IC50 value of 25.08 nmol/L. The expression of SDF-1alpha mRNA in lymph node stromal cells obtained from patients with NHL was decreased after treatment by triptolide at concentrations of 25 and 50 nmol/L for 24 h. Flow cytometry analysis showed that the CXCR4 expression on primary lymphoma cells were downregulated gradually in a dose-dependent manner following triptolide treatment. Chemotaxis assays revealed that the migration of freshly isolated lymphoma cells towards either rhSDF-1 or cultured lymph node stromal cells was markedly inhibited by the addition of triptolide in vitro, and the inhibition was dose-dependent., Conclusion: Triptolide can inhibit the proliferation of B-NHL cell line Raji cells. Moreover, triptolide is able to inhibit the migration of lymphoma cells via lymph nodes in vitro. The potential antitumor mechanisms of triptolide are related to the antiproliferative effect and the blockage of SDF-1/CXCR4 axis.

Aim: To investigate the effects of triptolide, a diterpenoid triepoxide from Tripterygium wilfordii Hook F (TWHF), on the co-stimulatory molecule expression and interleukin-12 (IL-12) production from THP-1 cells., Methods: THP-1 cells were differentiated into macrophage-like cells by Me2SO, and then cultured with IFN-gamma (500 kU/L) and lipopolysaccharide (LPS) (1 mg/L) with or without triptolide. The surface molecule expressions were analyzed on a FACScan flow cytometer. IL-12p40, IL-12p70 were assayed by ELISA., Results: Triptolide suppressed CD80 and CD86 expressions on IFN-gamma (500 kU/L) and LPS (1 mg/L) activated THP-1 cells at nontoxic dosages of 2.5-0.625 microg/L. Furthermore, the production of IL-12p40 and IL-12p70 were also significantly reduced in THP-1 cells exposed to triptolide., Conclusion: Triptolide impairs the antigen-presenting function by inhibiting CD80 and CD86 expressions and decreased IL-12p40 and IL-12p70 (bioactive form) productions from the activated THP-1 cells.

Aim: To investigate the therapeutic effect of the glucosides of Cheanomeles speciosa (GCS) on the collagen-induced arthritis (CIA) in mice., Methods: Mice were divided randomly into six groups, including normal, CIA, CIA+GCS (60, 120, and 240 mg/kg) and CIA plus glucosides of Tripterygium wilfordii (GTW) groups. CIA model was based on mice. The effect of GCS in CIA mice was measured by paw-swelling, arthritis scores, and histopathological assessment of synovium. Indices of thymus and spleens were measured. Thymocytes and splenocytes proliferation, activity of interleukin-1 (IL-1), and interleukin-2 (IL-2) were assayed by MTT and [(3)H]TdR method. The level of anti-collagen type II (CII) antibody in serum and prostaglandin E (PGE) in ankle were assayed by ELISA and ultraviolet spectrophotometer method, respectively., Results: The onset of paw-swelling was on d 24 after injection of emulsion. The peak of secondary inflammation appeared on d 36 and then declined after d 40. GCS and GTW significantly reduced paw-swelling and arthritis scores, reduced the increase of spleen indices of CIA mice, suppressed the ConA or LPS-induced thymocyte or spleen cell proliferation, and the production of IL-1 and IL-2 in CIA mice. GCS reduced the level of anti-CII antibody and PGE. Histological pathology analysis demonstrated that the synovium of CIA mice was hyperplastic, pannus was formed, and inflammatory cells infiltrated into synovium. The pathological changes were significantly reduced by GCS., Conclusion: GCS had anti-inflammatory effect on CIA mice, which might be related to the modification of the abnormal immunological function of CIA mice.

Aim: To investigate the global gene expression profile changes in Jurkat cells after triptolide treatment in order to find the possible triptolide targets., Methods: Jurkat cells were treated with or without triptolide 10 microg/L for 2 h. Total RNA were isolated and used as templates for reverse transcriptional labeling of fluorescent cDNA probes. High density DNA microarray chips with a set of 13,872 human genes/Ests were used to generate the expression profile of triptolide-treated or untreated control Jurkat cells by hybridizing with fluorescent labeled probes. Array image was acquired and analyzed with array analyzing software GeneSpring., Results: Triptolide significantly suppressed expression of 117 genes in Jurkat cells. Among these 117 genes, 30 % were Ests or genes without known functions, 13 % were transcription factors, 9 % were signal transduction pathway regulators, and 9 % were DNA binding proteins. Notably, the expression of mitogen-activated protein kinase kinase kinase kinase 5 (MAP kinase 5) and phosphoinositide-3-kinase (PI-3 kinase) was inhibited more than 100-fold. Moreover, the expression of genes involved in lipid transportation and metabolism was down-regulated by triptolide., Conclusion: High-density microarray provided an effective approach to identify drug targeting molecules. It is suggested that the widely known immune suppressive and antitumor effects of triptolide were mediated at least in part by suppression of MAP kinase and PI-3 kinase gene expression.

Ion channel plays a key role in maturation, capacitation and acrosome reaction of sperm. However, as it is difficult to record channel currents from a mature mammal sperm directly by patch-clamp technique, there were no basic data on the types and properties of the channels in human sperm until the method reconstituting the channels into bilayer was used. By reconstituting the channel proteins isolated from sperm membrane into phospholipid-forming bilayer, we have characterized several kinds of Ca2+-, Na+-, K+-, and Cl--permeable channels with different conductance and properties in human sperm membrane. To study the channels in spermatogenic cells is another approach to understand the ion-channels in mature sperm. The cell is used as a model to analyze the effects of male antifertility agents on Ca2+-channel. To date, several male contraceptives derived from Chinese traditional medicine have been attached worldwide interest, a lot of compounds have been purified from them, and the antifertility effects of some compounds were demonstrated. We studied the effects of gossypol and several compounds isolated from Tripterygium wilffordii on Ca2+ channel in mouse spermatogenic cells and found that each of them inhibited the channel and sperm acrosome reaction at a proximate concentration, suggesting that the inhibition of Ca2+ channels may be one mechanism of the antifertility effects of these contraceptives.

Aim: To study the effects of Tripterygium wilfordii (TII) and IL-10 on human leukocyte antigen-DR (HLA-DR) and CD80 expressions and IL-12 p40 subunit production and transcription of dendritic cells (DC) in human peripheral blood from healthy volunteers in vitro., Methods: DC were generated by culturing plastic-adherent peripheral blood mononuclear cells with GM-colony stimulating factor (GM-CSF), IL-4, and TNFalpha. The expressions of HLA-DR and CD80 were examined by flow cytometry after the cells were stained with immunofluorescence antibodies. Enzyme-linked immunosorbent assay and reverse transcriptase polymerase chain reaction analysis were used to detect IL-12 p40 protein level and mRNA transcription, respectively., Results: TII 5 - 20 mg/L and IL-10 50 - 200 microg/L greatly down-regulated the membrane expressions of HLA-DR and CD80 on DC in a concentration-dependent manner. IL-12 p40 production and mRNA transcription were also inhibited in DC both by TII and by IL-10., Conclusion: TII and IL-10 exert immunosuppressive role via inhibiting membrane expressions of HLA-DR and CD80 and synthesis of IL-12 p40 subunit in DC.

Aim: To observe the effects of tripcholorolide (T4) on inflammatory reaction of mouse alveolar macrophages., Methods: RT-PCR was used to investigate tumor necrosis factor alpha (TNF alpha), interleukin-1 beta (IL-1 beta), IL-6, IL-10, and inducible nitric-oxide synthase (iNOS) mRNA expression in alveolar macrophages after LPS 10 mg/L and T4 500 micrograms/L treatment. ELISA was used to detect TNF alpha, IL-1 beta, IL-6, and IL-10 protein expression. Nitrite was measured by Griess reaction., Results: TNF alpha, IL-1 beta, IL-6, IL-10, and nitrite increased in supernatants, when alveolar macrophages were stimulated by LPS 10 mg/L at 24 h. Both T4 500 micrograms/L and dexamethasone 100 mumol/L had inhibitory effects on the production of TNF alpha, IL-1 beta, IL-6, IL-10, and nitric oxide. The mRNA expression of TNF alpha, IL-6, IL-10, and iNOS increased at 5 h after LPS stimulation which was decreased on addition of T4 500 micrograms/L or dexamethasone 100 mumol/L. T4 had no effect on stability of LPS-induced mRNA expression in TNF alpha, IL-6, and IL-10., Conclusion: T4 had inhibitory effects on the expression of proinflammatory and antiinflammatory mediators.

Aim: To study the roles of Tripterygium wilfordii saponins (TII) and interleukin-10 (IL-10) on dendritic cells (DC)-induced experimental autoimmune thyroiditis (EAT)., Methods: We used mice as autoimmune thyroiditis model animals and divided them into 4 groups, namely DC group, TII group, IL-10 group, and control group. The level of thyroglobulin (Tg) antibody was assayed by ELISA. TNF beta production in the cultured supernatants and nitric oxide (NO) in the serum were measured by biological activation assay and Griess reaction, respectively. Tg-stimulated proliferation of lymphocytes was detected with MTT incorporation assay. The histopathological analysis of thyroid was carried out., Results: Tg-pulsed DC were able to induce EAT with increase in the concentration of TgAb in serum and lymphocytes infiltration in thyroid. After treatment with TII or IL-10, DC could not induce EAT with lower levels of TgAb and no lymphocyte infiltration. The concentration of NO in serum, TNF beta activation, and the proliferation of lymphocytes in response to thyroglobulin in TII or IL-10 group were lower than those in DC group., Conclusion: TII and IL-10 are able to strongly inhibit the ability of DC to induce experimental autoimmune thyroiditis.