Inhibitory effect of active ingredients of Tripterygium wilfordii Hook.F.on human carboxylesterases

OBJECTIVE The inhibitory effect of active ingredients of Tripterygium wilfordii Hook.F.(TWHF)(celastrol,triptolide,triptonide,wilforlide A,wilforgine and wilforine)on human carboxylester⁃ase 1(CES1)and CES2 was detected to investigate the herb-drug interactions(HDIs)of TWHF.METHODS Human liver microsomes catalysed hydrolysis of 2-(2-benzoyl-3-methoxyphenyl)benzothi⁃azole(BMBT)and fluorescein diacetate(FD)were used as the probe reaction to phenotype the activity of CES1 and CES2,respectively.The residual activities of CES1 and CES2 were detected by ultrahigh performance liquid chromatography(UPLC)after intervention with celastrol,triptolide,triptonide,wilforlide A,wilforgine and wilforine(100μmol·L^(-1)).Kinetics analysis,involving half inhibitory concentra⁃tion(IC_(50)),inhibition type and kinetic parameter(Ki),and in vitro-in vivo extrapolation(IVIVE),was carried out to predict the HDIs between these compounds and CES-metabolizing drugs.Molecular docking was performed to analyze the ligand-enzyme interaction.RESULTS Out of the six main con⁃stituents of TWHF,only celastrol exhibited strong inhibition towards both CES1 and CES2,with the inhibitory rates of 97.45%(P<0.05)and 95.62%(P<0.05),respectively.The IC_(50)was 9.95 and 4.02 mol·L^(-1),respectively,and the types of inhibition were all non-competitive inhibition.Based on the kinetics analysis,the Ki values were calculated to be 5.10 and 10.55μmol·L^(-1)for the inhibition of celastrol on CES1 and CES2,respectively.IVIVE indicated that celastrol might disturb the metabolic hydrolysis of clinical drugs in vivo by inhibiting CES1.Molecular docking results showed that hydrogen bonds and hydrophobic contacts contributed to the interaction of celastrol and CESs.CONCLUSION The inhibitory effect of celastrol on CES1 and CES2 might cause HDIs with clinical drugs hydrolysed by CESs.

In vitro propagation of a medicinal plant: Tripterygium wilfordii Hook f.

In this study a reliable protocol was developed for the establishment of commercial in vitro cultures of Tripterygium wilfordii Hook f.. Juvenile shoots from one-year-old elite plants were used as the source of explants. New axillary shoots were obtained after 30 days of culture on a MS medium supplemented with BAP （2.0 mg.L^-1） and NAA （0.1 mg.L^-1）. The optimal multiplication medium was a modified MS medium supplemented with BAP （1.0 mg.L^-1） and NAA （0.1 mg.L^-1）. This yielded a multiplication rate of 2.4 for each subculture. Slightly more than 92% of shoots rooted when cultured on a modified MS medium containing IBA （0.2 mg.L^-1） and activated charcoal （0.5 mg.L^-1）. Activated charcoal promoted both a strong and a high rooting rate during the rooting phase. Plantlets were transferred to pots for a short acclimatization stage in a greenhouse where 95% of the plantlets survived. This highly reproducible procedure can be adopted for large-scale propagation of T. wilfordii.

Isolation and Purification of Triptolide from the Leaves of Tripterygium wilfordii Hook F

Triptolide is an important active component of Tripterygium wilfordii Hook F (TWHF) and possesses anti-inflammatory, immunosuppressive, male anti-fertility, and anticancer properties. A new method combining different techniques, including solid-liquid extraction, liquid-liquid partition, column chromatography and high-speed counter-current chromatography (HSCCC) but avoiding the use of chloroform, was developed for the isolation and purification of triptolide from the leaves of TWHF. 48 mg of triptolide at 96.5% purity was obtained from 1 kg of air-dried leaves of TWHF.

Effect of the Multiglycoside of Tripterygium Wilfordii Hook f. (Tii) on Cornea Allograft Rejection Model in Rabbit

Purpose:Toexamine the effect of Tii treatment of cornea graft survival in a rab-bit model.Methods:Tii was administrated orally after eccentrical corneal transplantation.Survival times were determined by biomicroscopy.Cytotoxic T lymphocytes(CTL)and delayed-type hypersensitivity(DTH)responses to donor alloantigens were assessed at ady 16after heterotopic corneal grafts.Results:Administration of Tii reduced the incidence and prologed the graft sur-vival time.Both CTLand DTH responses to donor alloantigens were severely ed-pressed in hosts treated with Tii.However,combination of Tii and cyclosporine further enhanced the immunosuppressive effects described above.Conclusions:Tii is a potent immunosuppressant with the ability to prolong allo-graft survival in the rabbit penetrating keratoplasty model and may have coordi-native effects with CsA through different mechanisms.Further studies are neces-sary to define any potentially coordinative role in the prevention of allograft rejec-tion in human keratoplasty.Eye Science 1995;11:168-172.

A Novel Diterpenoid,11-O-β-D-glucopyranosylneo-triptophenolide, from Tripterygium wilfordii Hook. f

A new diterpenoid was isolated from the ethanolic extract of the dried root bark of Tripterygium wilfordii Hook.f. It is the first example of abietane diterpenoid glycoside isolated from Tripterygium wilfordii Hook.f. Its structure was identified to be 11-O-b-D-glucopyranosyl- neotriptophenolide based on spectral methods.

雷公藤内酯醇(T10)通过抑制脊髓内胶质细胞的激活改善大鼠CFA诱导的炎性痛的机制研究

目的:观察腹腔注射雷公藤内酯醇(Triptolide,T10)对完全弗氏佐剂(complete Freund’s adjuvant,CFA)诱导的大鼠慢性炎性痛行为的改善作用并探讨其机制。方法:SD大鼠随机分为四组,即Saline+Veh组、Saline+T10组、CFA+Veh组和CFA+T10组。后两组大鼠于右侧足底注射CFA制备大鼠慢性炎性痛模型,前两组则在相同部位注射生理盐水。第二组和第四组大鼠分别于造模前1 h给予T10腹腔注射,随后每12 h给予腹腔注射药物1次,持续用药7 d;第一组和第三组则以相同方式给予100 ml/kg的生理盐水作为对照。采用辐射热法和机械刺激法连续观察给药后大鼠的痛行为;应用免疫组织化学染色和Western Blot方法观察连续给予T10 3d和7 d后大鼠腰膨大平面脊髓背角内小胶质细胞和星形胶质细胞的活化程度;应用实时定量PCR(RT-PCR)方法观察连续给药7 d后大鼠腰5背根神经节内炎性因子,如白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1 beta,IL-1β)和肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)的表达变化。结果:(1)与CFA+Veh组相比,CFA+T10组大鼠机械缩足阈值及辐射热痛潜伏期显著下降(P<0.05),并持续到造模成功后7 d,表明腹腔注射T10可以明显改善CFA诱导的大鼠机械痛敏和辐射热痛敏。(2)免疫组织化学染色和Western Blot结果显示:造模后3 d和7 d后CFA+Veh组大鼠脊髓背角出现大量CFA诱导活化的小胶质细胞和星形胶质细胞;小胶质细胞标记物IBa-1和星形胶质细胞标记物GFAP的表达量分别在CFA造模后3 d和7 d显著增高(P<0.05),而给予T10 3 d和7 d后炎性痛大鼠腰膨大平面IBa-1和GFAP的活化程度均显著减轻(P<0.05)。(3)RT-PCR结果显示:与Saline+Veh组相比,CFA造模7 d后腰5背根神经节内炎性因子IL-1β、IL-6 mRNA和TNF-αmRNA的表达显著增高(P<0.05),腹腔注射T10 7 d后上述炎性因子的表达则显著降低(P<0.05)。结论:腹腔注射T10可以显著改善CFA诱导的大鼠慢性炎性痛,其机制可能与抑制脊髓背角小胶质细胞和星形胶质细胞的活化以及炎性因子IL-1β、IL-6和TNF-α的合成有关。

雷公藤多甙抑制EM患者腹腔巨噬细胞及相应细胞模型分泌TNF-α的实验研究

目的观察雷公藤多甙(TWP)对子宫内膜异位症(EM)患者腹腔巨噬细胞和小鼠腹腔活化巨噬细胞释放TNF-α的影响,以探讨TWP用于临床治疗EM的机制并建立细胞模型,为后续研究做准备。方法收集16例经腹腔镜检查确诊为EM患者腹腔液分离纯化巨噬细胞,并给予不同剂量的TWP进行共同孵育;同时分离纯化小鼠腹腔巨噬细胞,用脂多糖(LPS)激活以获得细胞模型,同样予TWP进行共同孵育。分别以间接MTT法和RT-PCR法在蛋白质水平和基因水平检测该药物影响激活状态巨噬细胞分泌TNF-α含量的变化。结果 EM患者腹腔液中TNF-α含量显著高于非EM妇女,在4～24 h后TWP(6.25～100μg/L)显著抑制了TNF-α的分泌。结论 TWP在蛋白质和基因水平均能有效抑制激活巨噬细胞分泌TNF-α。

Efficacy and Safety of Tripterygium wilfordii Hook F Versus Acitretin in Moderate to Severe Psoriasis Vulgaris：A Randomized Clinical Trial

Background：Few clinical trials have evaluated the efficacy and safety of Tripterygium wilfordii Hook F （TwHF） compared with acitretin in psoriasis.We aimed to compare the efficacy and safety of TwHF compared with acitretin in the treatment of moderate to severe psoriasis vulgaris.Methods：Adults with Psoriasis Area Severity Index （PASI） score ＞ 10 and psoriasis-affected body surface area ＞ 10％ were randomized into a TwHF （20 mg,3 times a day） or acitretin group （30 mg,once a day）.The treatment course lasted for 8 weeks.Patients were assessed at baseline and at 2,4,and 8 weeks.Laboratory tests were performed at baseline,week 4,and week 8.The data were analyzed using paired samples t-test or analysis of variance （ANOVA）.Results：A total of 115 patients was enrolled （58 TwHF; 57 acitretin）.The median PASI score improved in the TwHF group by 50.4％ and in the acitretin group by 42.7％.There was no significant difference in median PASI improvement between two groups at 2,4,and 8 weeks.There was also no significant difference in PASI 25,PASI 50,PASI 75,and PASI 90 response between the two groups at 2,4,and 8 weeks.There was a significant increase in the level of aspartate transaminase and triglycerides in the TwHF group （P =0.026 and P =0.011,respectively）.In the acitretin group,there was a significant increase in the level of alanine transaminase,cholesterol,and high-density lipoprotein （P =0.030,P ＜ 0.01,and P ＜ 0.01,respectively）.Conclusions：There was no significant difference in treatment efficacy between the TwHF and acitretin groups within 8 weeks,but there were fewer treatment-related adverse events in the TwHF group.

Effects of triptolide on hippocampal microglial cells and astrocytes in the APP/PS1 double transgenic mouse model of Alzheimer＇s disease

The principal pathology of Alzheimer＇s disease includes neuronal extracellular deposition of amyloid-beta peptides and formation of senile pl aques, which in turn induce neuroinflammation in the brain. Triptolide, a natural extract from the vine-like herb Tripterygium wilfordii Hook F, has potent anti-inflammatory and immunosuppressive efficacy. Therefore, we determined if triptolide can inhibit activation and proliferation of microglial cells and astrocytes in the APP/PS1 double transgenic mouse model of Alzheimer＇s disease. We used 1 or 5 μg/kg/d triptolide to treat APP/PS1 double transgenic mice （aged 4-4.5 months） for 45 days. Unbiased stereology analysis found that triptolide dose-dependent- ly reduced the total number of microglial cells, and transformed microglial cells into the resting state. Further, triptolide （5 μg/kg/d） also reduced the total number of hippocampal astrocytes. Our in vivo test results indicate that triptolide suppresses activation and proliferation of microglial cells and astrocytes in the hippocampus of APP/PS 1 double transgenic mice with Alzheimer＇s disease.

Use of Tripterygium wilfordii Hook F for immune-mediated inflammatory diseases: progress and future prospects

Tripterygium wilfordii Hook F has significant anti-inflammatory and immunosuppressive properties and is widely used for treating autoimmune and inflammatory diseases such as rheumatoid arthritis, systemic lupus erythematosus, and kidney disease, especially in traditional Chinese medicine. The mechanisms underlying its effects may be diverse but they remain unclear, and its toxicity and side effects limit its wider clinical application. This review summarizes the clinical application of Tripterygium wilfordii Hook F in recent years, as well as the results of studies into its mechanisms and toxicity, to provide a reference for its future clinical application.

Gene expression profile analyses of mice livers injured by Leigongteng

AIM： To analyze the gene expression profiles of mice livers injured by Leigongteng and explore the relationship between the differentially expressed genes and liver damage. METHODS： The experimental mice were randomly divided into a control group and a liver-injured group in which the mice were administrated 33 μγ, of triptolide/ kg per day for 30 d. Liver mRNAs were extracted from animals in both groups and were reverse-transcribed to cDNA with dUTP labeled by different fluorescence （Cy3, Cy5） as hybridization probes. The mixed probes were hybridized with oligonucleotide microarray chips. The fluorescent signal results were acquired by scanner and analyzed with software. RESULTS： Among the 35852 target genes, 29 genes were found to be significantly differentially expressed, with 20 genes up-regulated and 9 genes down-regulated. The reliability of the differentially expressed genes was validated by RT-PCR experiments of 5 randomly selected differentially expressed genes. CONCLUSION： Based on the biological functions of the differentially expressed genes, it is obvious that the occurrence and development of liver damage induced by Leigongteng in mice are highly associated with immune response, metabolism, apoptosis and the cell skeleton of liver cells. This might be important for elucidating the regulatory network of gene expression associated with liver damage and it may also be important for discovering the pathogenic mechanisms of liver damage induced by Leigongteng.

Decreased bone mineral density in female patients with systemic lupus erythematosus after long term administration of Tripterygium Wilfordii Hook F

Objective To investigate the possible bone changes in female patients with systemic lupus erythematosus (SLE) induced by long term administration of Tripterygium Wilfordii Hook F (TW) Methods 70 female SLE patients were divided into 4 groups accordiog to their drug history: SLE disease control group, corticosteroids treatment group, TW treatment group, and both corticosteroids and TW treatment group Bone mineral density (BMD) of the lumbar spine 2-4 and biochemical markers of bone turnover were studied Results Long term administration of TW could significantly decrease BMD levels in female SLE patients ( P <0 05) The patients receiving TW for more than 5 years had significantly lower BMD levels compared with those for less than 5 years The degree of decreased BMD induced by TW was less severe compared with that of prednisone No significant differences were observed in the biochemical markers of bone turnover among four groups ( P >0 05) Conclusion Long term administration of TW could decrease BMD levels in women Osteoporosis may be an important problem for SLE patients treated with TW

ULTRASTRUCTURAL OBSERVATION ON MACROPHAGE-LYMPHOCYTE INTERACTIONS IN SEMEN FROM TRIPTERYGIUM WILFORDII HOOK.F. TAKERS

Sixty-nine specimens from Tripterygium Wilfordii Hook.f. (TWH) users were investigated by electron microscopy. No macrophages were demonstrated in the 21 specimens collected prior to the administration of TWH. However, it was found in 23 out of the 48 semen specimens obtained following the TWH administration. The macrophages were functionally active as shown by the presence of a large number of cytoplasmic processes and pseudopodia on the surface, and primary and secondary lysosomes in the cytoplasm. The macrophages phagocytized sperm debris and degenerated or dead spermatids with formation of specific phagosomes. Around those macrophages, lymphocytes were commonly noted. The cytoplasmic processes of the two cell types could come into contact or even fuse with each other, leading to tight junction-like structure; in some of the contacts, the plasma membranes were found dissolved so as to form direct cytoplasmic linkage.

PROSTATE CANCER BONE-METASTASIS IS INHIBITEDBY CELASTROL, AN ACTIVE CONSTITUENT OF TRIPTERYGIUM WILFORDII HOOK.F.(雷公藤)

Prostate cancer(PCa)is one of the most common malignant tumours and a leading cause of cancer deaths.Treatment failure of PCa is often due to bone-metastasis.Celastrol,an active constituent of the roots of Tripterygium wilfordii,has shown antitumour effects in previous studies in accordance with its use in TCM.Here we report for the first time an in-depth study of the effects of celastrol on

Combination of Tripterygium Wilfordii Hook F With Antiretroviral Therapy Delayed Viral Rebound in A Patient of Acute HIV-1 Infection

Prolonged antiretroviral therapy(ART)-free remission post-treatment has been observed and reported in human immunodeficiency virus-1(HIV-1)infection.The primary factors for such achievement have been linked to the transcriptional inactivation of HIV-1 DNA and immune recovery.Here,we reported a patient with acute HIV-1 infection who immediately received intensified quadruple ART.At month 9(M9),an old CFDA-approved immuno-suppressive herbal medicine,Tripterygium Wilfordii Hook F(TwHF),was used in addition to ART.The patient was closely monitored.Virological and immunological tests as well as transcriptome analysis were carried out at each visit.The results showed that TwHF reduced serum IP-10 level and inhibited T cell activation.Both ART and TwHF were discontinued in M24,and levels of peripheral blood HIV-1 RNA and DNA remained suppressed for consecutive 12months.With transcriptome analysis,we found pattern changes linking immuno-activation and amino acid metabolism with viral suppression and rebound.This indicates that the intentional suppression of immuno-activation is a promising approach for a functional cure of HIV-1 infection.

Effects of GTW treatment on proteinuria and acute glomerular immune lesion in experimental mesangial proliferative glomerulonephritis

To examine the effect of multi-glycoside of Tripterygium wilfordii Hook. f. （GTW） on proteinuria and acute glomerular immune lesion in experimental mesangial proliferative glomerulonephritis （MsPGN） induced by anti-Thyl. 1 monoelonal antibody （mAb） 1-22-3. The reversible model of MsPGN with anti-Thyl. 1 mAb 1-22-3 was established. After 7 days of oral treatment with GTW （ 100 mg/kg per day） and vehicle （distilled water, 5 ml/kg per day）, its effects on proteinuria, renal functions, mesangial morphological change, glomerular macrophage accumulation, and mRNA expressions of cytokines （PDGF-BB and MCP-1 ） were evaluated by light microscope （LM）, immunofluorescence （IF）, and reverse transcription polymerase chain reaction （RT-PCR）. It was found that GTW ameliorated proteinuria （on day 3 and day 7）, mesangial proliferation （total cell number, matrix expansion, a-smooth muscle actin expression, and collagen type Ⅰ expression） and macrophage accumulation （ED3^＋ ） in experimental MsPGN. In addition, GTW significantly suppressed the increased mRNA expressions for MCP-1 （67.6% to eontrol group, P 〈 0.01） together with the tendency to reduce the expression of PDGF （24.44% to control group） on day 7. It is concluded that GTW can not only decrease proteinuria, but also ameliorate acute mesangial alterations and glomerular activated macrophage accumulation probably by reduction of cytokines. These data indicate that GTW is an effective agent for early MsPGN.

HPLC法测定雷公藤及其片剂中雷公藤红素

目的建立雷公藤及其片剂中雷公藤红素的HPLC测定方法.方法 HPLC外标法.Agilent Zorbax SC-C18柱为色谱柱,甲醇-1%醋酸溶液(87:13)为流动相,检测波长为425 nm,体积流量为1.0 mL/min.结果雷公藤红素线性范围为40.96～204.8μg/mL(r=0.999 6),药材的平均回收率为98.37%,RSD为1.01%(n=9),制剂的平均回收率为98.59%,RSD为1.18%(n=9).结论本方法操作简便、准确,可作为雷公藤类药物中雷公藤红素的定量分析方法.

雷公藤有效成分制备的研究进展

简要介绍了近年来有关中药雷公藤有效成分的制备方法及研究进展 。

系膜增殖性肾小球肾炎的新疗法——雷公藤多甙新治疗方案的疗效观察

目的：应用双倍剂量雷公藤多甙（ＴⅡ）继以间歇用药维持治疗方案治疗不同临床病理类型的原发性系膜增殖性肾小球肾炎，前瞻性观察其疗效及副反应，确定此一新疗法的临床价值。方法：７９例无氮质血症的原发性系膜增殖性肾小球肾炎患者（ＩｇＡＮ４７例，ＩｇＭＮ１５例，ＭｓＰＧＮ１７例）在连续服用ＴⅡ２ｍｇ／（ｋｇ·ｄ）４周后改为间歇疗法，每日２ｍｇ／ｋｇ，服２周停２周，交替进行，平均随访１９．３周，观察蛋白尿、血尿变化及副反应。结果：４７例尿检异常型ＩｇＡＮ患者中，１９例达临床缓解（４０．５％），５例改善（１０．６％），２３例无效（４８．９％）。其中表现为单纯蛋白尿者疗效最佳，其次为单纯血尿者，蛋白尿伴血尿者疗效最差。１７例ＭｓＰＧＮ患者中，１１例达临床缓解（６４．７％），６例无效（３５．３％）。同样以单纯蛋白尿者疗效最佳，其次为蛋白尿伴血尿者，单纯血尿疗效最差。１５例ＩｇＭＮ患者中，８例达临床缓解（５３．３％），３例改善（２０％），４例无效（２６．７％）。１９例以往曾用足量激素治疗无效的患者中有１２例（６３．２％）经双倍剂量ＴⅡ治疗后获缓解。临床缓解的３８例患者中，缓解时间在４周以内者２６例（６８．４％），６周内者３２例（?

“五子四物瓜石汤”对雷公藤多苷所致雄性大鼠生殖系统毒性的对抗作用及其机制研究

目的 研究自拟中药复方“五子四物瓜石汤”(WT)对雷公藤多苷 (GTW)所致雄性大鼠生殖系统毒性的对抗作用及其机制。方法 雄性大鼠分别 ig大、中、小剂量的 WT与固定剂量的 GTW,连续给药 80 d,以单独 igGTW或 NS为对照 ,观察各组雄性大鼠睾丸脏器指数、精子数、精子活率、睾丸病理结构的变化 ,并用免疫放射分析法 (IRMA)测定各组雄性大鼠血清性激素水平。结果 与单用 GTW组比较 ,大或中剂量 WT可使睾丸重量明显增加 ,精子数明显升高 ,精子活率显著提高 ,其中大剂量 WT可完全对抗 GTW对生殖上皮的损害 ,使睾丸结构完全正常 ,血清睾酮维持于正常水平。结论 “五子四物瓜石汤”能显著对抗雷公藤多苷对雄性大鼠生殖系统的毒性作用 ,其对抗作用与 WT保护睾丸生精上皮免受 GTW的损害 。