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.

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

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.

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.

“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 。

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（?

双倍剂量雷公藤多甙治疗原发性肾病综合征的近期疗效

目的：首次应用双倍剂量雷公藤多甙（ＴⅡ）治疗原发性肾病综合征，前瞻性观察其疗效。方法：１８例单纯肾病综合征患者（８例ＩｇＡＮ，４例ＩｇＭＮ，６例ＭｓＰＧＮ）均服用ＴⅡ２ｍｇ／（ｋｇ．ｄ）４周，观察蛋白尿变化及副反应。结果：１５例肾病综合征获得完全缓解，总缓解率达８３３％，其中ＩｇＡＮ（７例），ＩｇＭＮ（３例），ＭｓＰＧＮ（５例）的缓解率分别为８７５％，８３３％，７５０％。１５例中１２例在服药２周内获得缓解，部分对皮质激素不敏感者双倍剂量ＴⅡ仍然有效，无效仅２例。尿蛋白平板电泳分析标明对ＴⅡ敏感者尿蛋白呈高度选择性，而对ＴⅡ不敏感者含有较多大分子及小分子量蛋白质。所有患者均能耐受双倍剂量ＴⅡ，消化道反应及对肝细胞的损伤作用轻微，无一例出现严重白细胞减少。分别有２例患者在ＴⅡ迅速减量及继发上呼吸道感染后肾病综合征复发，但重新采用大剂量ＴⅡ或延长治疗时间仍有效。结论：双倍剂量ＴⅡ对临床表现为单纯肾病综合征，组织学病变为系膜增生的多种肾小球疾病具有良好的疗效。

雷公藤与昆明山海棠采样调查报告

目的:通过采样调查,获取雷公藤与昆明山海棠野生资源现状、种植、商品流通等方面的资料。方法:在全国13个样地采集样品,比较雷公藤和昆明山海棠的形态特征与生长习性的差异;调查雷公藤与昆明山海棠的野生资源现状、种植信息、民间混淆种及不同市售药材的商品特征。结果:(1)雷公藤和昆明山海棠主要采集于林下或林地边缘,光照和水分对雷公藤属植物的分布有一定的影响;(2)雷公藤与昆明山海棠的野生资源量正在萎缩,两种植物种植历史较短,种植技术水平较低;(3)雷公藤的采收加工尚缺乏规范,市售药材将雷公藤属植物的根、根茎、茎均作商品药材销售,饮片规格多样。结论:雷公藤与昆明山海棠的野生资源正在萎缩,人工种植、采收加工及市售药材的管理等方面的研究尚需规范深入的开展。

雷公藤诱导细胞凋亡的研究进展

雷公藤 Tripterygium wilfordii提取物在临床上常用于治疗类风湿关节炎等多种疾病 ,其疗效已被广泛认可。近 5年来国内外学者从其提取物诱导细胞凋亡的角度 ,对其药理机制进行了细胞分子水平的探讨 。

雷公藤多甙抑制小鼠腹腔巨噬细胞产生一氧化氮

雷公藤多甙（ｐｏｌｙｇｌｙｃｏｓｉｄｅｏｆＴｒｉｐｔｅｒｙｇｉｕｍｗｉｌｆｏｒｄｉＨｏｏｋ．ｆ．，ＴＷＰ）临床广泛用于治疗类风湿性关节炎等多种自身免疫性疾病，既可抑制细胞免疫又可抑制体液免疫［１］．但其在免疫抑制过程中对一氧化氮合酶（ＮＯＳ）的影响却...

同种异体肾移植术后长期应用雷公藤多苷的临床观察

目的：观察肾移植术后长期应用雷公藤多苷（TⅡ）的临床效果。方法：对223例同期肾移植受者进行对照性临床研究，随机分为TⅡ组（n=121）和对照组（n=102），另根据TⅡ剂量的不同将TⅡ组121例患者分为常规剂量TⅡ组（n=82）和双倍剂量TⅡ组（n=39）。以糖皮质激素、环孢素和硫唑嘌呤作为防治排斥的“三联”免疫抑制基础药物。各组患者均无并发感染、环孢素肾中毒和手术并发症。各组之间的性别、年龄、透析时间、冷缺血和热缺血时间、淋巴细胞毒性试验、群体反应性抗体水平均非常接近。结果：TⅡ组与对照组术后3个月内经病理证实的急性排斥发生率有明显差异，分别为4．1％和26．5％。TⅡ双倍剂量组在术后3个月内无一例发生急性排斥。TⅡ组中发生急性排斥的病理程度也较对照组轻。根据Banff分类，TⅡ组5例患者细胞性排斥为中度急性排斥（ⅡA级）；对照组27例急性排斥中，重度急性排斥（Ⅲ级）和中度急性排斥（ⅡB级）各11例。两组之间慢性移植肾肾病（CAN）的发生率分别为7．4％和12．7％。TⅡ组移植肾5年存活率明显高于对照组（90．9％郴77．5％。P〈0．01）。TⅡ组大多数受者5年内肾功能正常，肾功能异常（SCr≥132．6μmol／L）的发生率明显低于对照组，分别为33．6％和61．8％。对血肌酐倒数到达透析需要的时间（1／SCr为0．1）进行预测，即1／SCr值由0．5～0．1，预期到达的时间（月），TⅡ组和对照组之间的预期值分别为0．5：90．14和42．67；0．2：164．71和66．79；0．1：186．91和78．08。结论：肾移植术后长期应用TⅡ能有效地降低肾移植术后急性排斥发生率，减轻排斥反应的病理程度，降低CAN的发生率，保持移植肾功能长期稳定。雷公藤多苷不良反应轻，适合于长期用药。

雷公藤免疫抑制作用研究进展

传统中药雷公藤Tripterygium wilfordii为卫矛科雷公藤属植物,在我国的使用已有几千年的历史。其化学成分复杂,具有多种药理活性,是目前国内外研究的热点天然药物之一。近年来,雷公藤的各种提取物及有效成分在各类免疫性疾病中得到了广泛应用并取得了很好的疗效。现综述近几年国内外对雷公藤免疫抑制活性成分、免疫抑制机制研究的进展和临床应用的状况,并认为雷公藤作为一种极有潜力的免疫抑制药物,其应用前景十分广阔,值得对其进行深入的开发研究。

雷公藤多甙治疗肾移植后西罗莫司相关蛋白尿的临床研究

目的：已发现西罗莫司可导致肾移植患者蛋白尿。文中对应用西罗莫司联合霉酚酸酯和激素“三联”免疫抑制方案的肾移植受者进行临床观察，探讨雷公藤多甙（T Ⅱ）治疗西罗莫司相关蛋白尿的安全性和有效性。方法选择全军肾脏病研究所2009年4月至2012年12月间176例接受西罗莫司治疗的尸肾移植受者中，西罗莫司治疗6个月内出现尿蛋白的肾移植受者52例，包括9例新接受西罗莫司治疗和43例由钙调磷酸酶抑制剂（ calcineurin inhibitors， CNIs）更换为西罗莫司治疗患者。所有患者平均尿蛋白水平（4．5±2．8）g／24 h。将52例受者根据不同治疗方案纳入雷公藤治疗组（雷公藤组，n＝27）和缬沙坦组（n＝25），持续观察12个月。雷公藤组应用常规剂量雷公藤多甙[1 mg／（kg＆#183; d），雷公藤多甙含雷公藤内酯醇30μg／（10 mg＆#183;片）]；缬沙坦组剂量为80～160 mg／d。原西罗莫司联合霉酚酸酯和激素“三联”免疫抑制方案不变，根据西罗莫司目标浓度6～10 ng／mL（ ELASA方法）调整西罗莫司剂量；霉酚酸酯750 mg，2次／d，根据霉酚酸酯AUC 0－12水平（35～45 mg＆#183; h／L）调整剂量。疗效评估包括：①完全缓解，尿蛋白下降＞50％；②部分缓解，尿蛋白下降20％～50％；③无效，尿蛋白下降＜20％。结果随访12个月，2组患者的尿蛋白水平均显著下降，雷公藤组和缬沙坦组总有效率分别为96．3％和92．0％，雷公藤组尿蛋白下降较比缬沙坦组更加明显（ P＜0．01）。雷公藤组总胆固醇和三酰甘油水平均明显低于缬沙坦组（ P＜0．05）；缬沙坦组总胆固醇和三酰甘油水平有所升高，分别为[（6．60±0．22）mmol／L vs （7．11±1．13）mmol／L]和[（2．47±1．48）mmol／L vs （2．49±0．32）mmol／L]。雷公藤组血清清蛋白水平明显高于缬沙坦组[（41．1±1．2）g／L vs （37．9±4．2）g／L，P＜0．05]。缬沙坦组不良反应的发生率明显高于雷公藤组。结论雷公藤多甙和缬沙坦均可降低西罗莫司相关的蛋白尿，雷公藤组尿蛋白下降的幅度比缬沙坦组更加明显。

雷公藤优良单株选择技术(Ⅰ)——雷公藤种内变异、生物量模型和选择性状的研究

野生雷公藤种内存在丰富的变异，变异程度：枝数量〉冠幅〉主藤长〉地径，各表型间存在极显著的相关性；人工栽培的雷公藤植株总生物量构成：根52．75％、茎37．67％、叶9．58％，根是主要收获部位；雷公藤根系的平均含水率为43．88％，下坡〉中坡〉上坡；根系生物量组成以粗根为主，在不同坡位上均呈现粗根〉中根〉细根的规律；通过逐步回归，建立了雷公藤植株总生物量与枝数量、地径、平均藤径、平均藤长的回归模型，相关系数r=0．9807，说明以雷公藤枝数量、地径、平均藤径、平均藤长为选择因子开展优良单株选择，可以实现以总生物量为最终目标的选择效果．

临床高剂量雷公藤多苷对幼年大鼠生育能力的影响

目的探讨儿科临床高剂量(1.5mg/㎏)雷公藤多苷(GTW)对幼年大鼠生育能力的影响。方法雄性、雌性SD幼鼠各50只,均随机分为空白组与GTW组,每组25只,GTW按每日9mg/kg给药,用药12周后,雄鼠按1:1与健康成年雌性大鼠合笼,雌鼠按2:1与健康成年雄鼠合笼,分3次合笼,每次合笼时间为2周,合笼后观察雌鼠受孕率、仔鼠离乳存活率。结果雌、雄鼠每次合笼后GTW组受孕率、离乳存活率与空白组比较差异均无统计学意义。结论临床高剂量GTW对幼年大鼠发育为成年大鼠后生育能力和所生仔鼠的生长发育未见明显影响。