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.

UItrastructural Observation on Rat Spermatozoa Treatedwith Tripchlorolide(T_4) from Tripterygium wilfordii

Adult male rats were treated orally with monomer T_4(Tripchlorolide) isolated from Tripterygium wilfordii, at the dose of 50μg/kg/day,6 days/week for6 weeks.Ultrathin section and freeze etching replica of seminiferous tubules and epididymal spermatozoa were examined with elec-tron microscope. The results showed that the spemiogenesis was inhibited T_4 in seminiferous tu-bules. However,the damage and disruption of the spermatozoa were more serious in the epididymis.Damage of the structure and function ot microtubule and microfilament may be the chief reason for sperm damage. Sperm membrane was also very sensitive to the treatiment of monomer T_4.

Progress of Botanical Resources, Chemical Ingredients and Pharmacolo- gical Study of Tripterygium Wilfordii

Tripterygium Wilfordii Hooks. f (TW) also called yellow vine, yellow vine wood, yellow wax vine, breakgut herb etc., belongs to Galactia genus, its medicinal parts mostly belong to the wooden part of the double-layer decortical root. Traditional Chinese medicine (TCM) holds that it has the function of clearing Heat and removing toxin, dispelling Wind to fr- ee collateral, relaxing tendon to activate blood circulation, subsiding swelling and analgesia, insecticide and hemostasis. The studies on botanical resources, chemical ingredients and pharmacology of TW are briefly introduced as follows.

Tripterygium Wilfordii： Structure，Chemistry and Activityof Male Antifertility Constituents

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CLINICAL STUDY ON EFFECT OF TRIPTERYGIUM WILFORDII HOOK. F. GLYCOSIDES ON UTERINE LEIOMYOMA

Objective To observe the therapeutic effect and side reactions of Tripterygium wilfordii Hook,f. ( GTW) glycosides on patients with uterine leiomyomas. Methods 65 normally cycling women with symptomatic uterine leiomyomas received 40mg daily dose GTW for 3 to 6 months. Baseline ultrasound tests were obtained to evaluate the sizes of myomas and uterus, then repeated three and six months after treatment. Blood samples were collected to determine the hormonal levels of in the mid-follicular and mid-luteal phases of the menstrual cycles before GTW therapy and at 3~4 months and 5~6 months after treatment. Results Significant decrease in leiomy-oma volume was shown in 39 of 65 (60% ) and 28 of 40 ( 70% ) patients after 3~4 months and 5~6 months of treatment, respectively. The decrease of the volume of leiomyoma was time-dependent as while 27. 84% and 51.60% in 3~4 months and 5 ~ 6months, respectively. 25 of 65 patients had amenorrhea during the course of treatment. Tripterygium wilfordii glycosides treatment induced a significant increase in LH and FSH levels (P < 0. 01) as compared with pretreatment values. In contrary, a significant decrease in E2 and P levels (P <0. 05) was found, but no changes were observed in T and PRL levels after treatment. Conclusion Tripterygium wilfordii might serve as an effective therapeutic agent for leiomyomas with fewer side effects. A reversible inhibitory effect on the ovary by Tripterygium wilfordii glycosides may be one of the mechanisms of Tripterygium wilfordii in decreasing leiomyoma volume.

16-Hydroxytriptolide" An Active Compound from Tripterygium wilfordii

From the dried roots and leaves of Tripterygium wilfordii^(1.2),a new diter- penoid triepoxide,16-hydroxytriptolide was isolated,and its structure and stereochemistry elucidated as 16-(S)-hydroxy-triptolide on the basis of spectral data(IR, MS, UV,~1HNMR,^(13)CNMR,2d-NMR,Selective Long-range DEPT)and x-ray crystallographic analysis.This compound showed definite antiinflammatory action,strong immunosuppressive and antifertile activities.In addition,a known compound,triptolide was also isolated and all the spectral signals of^1 HNMR and ^(13)CNMR were assigned.

Immunosuppressive terpenoids from Tripterygium wilfordii

Two new terpenes,triptobenzene P（1）and wilforone（2）were isolated from Tripterygium wilfordii,as well as 10 known terpenes.Their structures were elucidated by spectroscopic methods.Compounds 2-4,8,10,and 11 showed significant immunosuppressive activities.

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.

Three new sesquiterpene alkaloids from the root of Tripterygium wilfordii

Three new sesquiterpene alkaloids,1-desacetylwilforgine（1）,1-desacetylwilforine（2）,and 9′-hydroxy-2-nicotinoylwilforine （3） were isolated from the roots of Tripterygium wilfordii Hook f.,along with six known alkaloids.Their structures were established on the basis of spectral analysis.

A new triterpenoid from the roots of Tripterygium wilfordii

A new triterpenoid 3,4,6-trihydroxy-2-oxo-1(10),3,5,7-tetraen-23,24-nor-D:A-friedooleana-29-oic acid,as well as twelve known terpenes were isolated from the roots of Tripterygium wilfordii.Its structure was established on the basis of spectroscopic methods.

Two new terpenes from Tripterygium wilfordii

Two new terpenes,wilfornine H(1) and triptobenzene Q(2) were isolated from Tripterygium wilfordii,as well as 11 terpenes. Their structures were elucidated by spectroscopic methods.Compounds 5,9-13 showed significant immunosuppressive activities.

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.

Triterpenoids from Tripterygium wilfordii

A triterpenoid named 3β, 22α, 30-trihydroxy-urs-12-en (1) was isolated from the ethanol extract of Tripterygium wilfordii together with four known compounds. The structure of (1), which was for the first time isolated from nature, was established by spectral analysis and by comparison with that of the relative compound.

A Brief Review and Prospect of Treatment of Proteinuria by Tripterygium Wilfordii

Tripterygium wilfordii (TW) was reported by LI Lei-shi, et al in 1981 to have marked effect in reducing proteinuria when it was used to treat glomerular nephritis, and the conclusion they worked out has been now accepted by most of nephrologists in 20-year experimental studies and wide application in clinical practice. Its mechanism, adverse reaction and mutagenesis effects, and dosage in clinical application in the recent years are now briefly reviewed in this paper.

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.

Observations on Therapeutic Effects of Huangdan Decoctionand Tripterygium Wilfordii Compound Tablet on Membranous Glomerulonephritis in Rats

The model of membranous glomerulonephritis(MGN)in rats was successfully established using self-made cationic bovine serum albumin(C-BSA)and treated with Huangdan Decoction (HDD) and Tripterygium Wilfordii Co.tablet(TW).Results indicated that the levels of urinary protein,blood urea nitrogen(BUN)and serum creatinine(Scr) in treated groups(groups A,B and C)were significantly decreased as compared with the control group(group D)(PM0.01).By light and electron microscope and immunofluorescent technique,the damage to kidney in groups A,B and C was found much milder than that in group D with lesion in group A being slightest.These findings suggest that HDD and TW may alleviate the pathological lesions of MGN,prevent or retard its progression,and have remarkable therapeutic effects on MGN.

Comparative Potency of Seven Compounds Isolated from Tripterygium Wilfordii on Cultred Rat Leydig Cells and Sertoli Cells

The damaging effect of seven compounds isolated from Triplelygium Wilfordii Hook.F were tested on the rat Leydig cells and Sertoli cells in culture. The reslllts showed thai 10 μg/ ml orthosphenic acid (TW2), 10μg/ ml 39,22α -dihydroxy-△12 ursen- 30-oic acid(TW5), 5.0 μg/ ml 39,22a--dihydroxy-△12-oleallen--29-oic acid(TW6), 5.0 μg/ ml salaspermic acid(TW7), 0.5μg/ ml celastrol(TW27) alld 1.25μg/ ml polpunonic acid (TW28) Ivere all detrimental to Leydig cells. and Sertoli cells. Holvever, 30 μg/ ml wilforlide A (TW1) did not affect the viability of Leydig cells and Sertoli cells significantly.The most toxic compounds was TW27, and the leasl,TW1.These results suggested that the-COOH at the C-20 site of these triterpenoids may be the functional chemical group responsible for the damaging effect on the Leydig cells and Sertoli. cells.

Identification of Genes Involved in Celastrol Biosynthesis by Comparative Transcriptome Analysis in Tripterygium wilfordii

Tripterygium wilfordii has been renowned mostly because of the anticancer effects of its root extracts,which is partly ascribed to the presence of celastrol,a pentacyclic triterpenoid,as one of the main active components.Celastrol also has recently been reported as an effective prodrug in the treatment of obesity.Despite the promising activities,the pathway leading to celastrol biosynthesis,especially cytochrome P450(CYP)enzyme(s)that occur in its downstream steps,are largely unknown.This study conducted a comparative analysis of the T.wilfordii transcriptome derived from its root and leaf tissues.Differential gene expression analysis identified a number of root-specific CYP genes.Further phylogenetic analysis suggested specific family members of CYPs that may participate in the late steps during celastrol biosynthesis.Root-specific transcription factors(TFs)that may play regulatory roles in celastrol biosynthesis were also discussed.This genetic resource will aid in isolating the celastrol biosynthetic genes as well as engineering the celastrol biosynthesis pathway.

The Effect of Tripterygium Wilfordii Monomer T4 on Rat Spermatid Nuclear Protein Transition

Rat testis elongating spermatids and epididymal sperms were collected after 7 weeks of treatment with Tripterygium wilfordii monomer T4.Total nuclear basic protein(TNBP)was extracted from the elongating spermatid nuclei and the sperm nuclei isolated by sonication.Polyacrylamide gel electrophoresis has been used to separate the TNBP and individual proteins were quantified by scanning microdensitometry.It was found that the content of protamine was reduced and the TH(Total Histones)/RP(Rat Protamine)ratios were increased following treatment in the testis elongating spermatids, and same result was found in the epididymal sperms.These results suggest that the interruption of nuclear protein transition of testis spermatids induced by T4 might cause aberrant epididymal sperm nuclear protein and lead to infertility.The relationship between protamine and fertility was discussed.