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.

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.

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.

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.

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.

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.

HISTOCHEMICAL EFFECT OF THE MULTIGLYCOSIDES OF TRIPTERYGIUM WILFORDII HOOK.F.(GTW)ON THE TESTIS AND EPIDIDYMIS OF RATS

SD Twenty adult male rats were equally divided into a contro1 and an experimcntal group.GTW,10 mg/kg per day,was given to the experimental rats through gastric gavage 6 days a week for 8 weeks.To the control rats,the same amount of vehicle was given The animals were sacrificed when they became infertile and the fol1owing parameters of the testis and epididymis were examined:l)DNA(Feulgen′s method),2)RNA(Brachet′s tech-uique),3)LDH-X(Lojda′s method),4)ATPase(Wachstein′s procedure),5)Succinate dehydrc-genase(SDH,Pearson's method),6)Non-specific esterase(NSE,Lojda's technique)and 7)PAS reaction.Routine examination of the epididymal spermatozoa was also carried out.

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.

Three new sesquiterpene alkaloids from the root of Tripterygium wilfordii

三新 sesquiterpene 碱， 1-desacetylwilforgine (1 ) ， 1-desacetylwilforine (2 ) ，和 9 鈥 ? hydroxy-2-nicotinoylwilforine (3 ) 从 Tripterygium wilfordii 钩 f. 的根被孤立，与六已知的碱一起。他们的结构被建立根据光谱分析。

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.

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

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 leiomyoma 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. O1 ) as compared with pretreatment values, hi 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.

Inflammatory and Immunomodulatory Effects of Tripterygium wilfordii Multiglycoside in Mouse Models of Psoriasis Keratinocytes

Objective:To determine the role of Tripterygium wilfordii multiglycoside(TGW) in the treatment of psoriatic dermatitis from a cellular immunological perspective.Methods:Mouse models of psoriatic dermatitis were established by imiquimod(IMQ).Twelve male BALB/c mice were assigned to MQ or IMQ+TGW groups according to a random number table.Histopathological changes in vivo were assessed by hematoxylin and eosin staining.Ratios of immune cells and cytokines in mice,as well as PAM212 cell proliferation in vitro were assessed by flow cytometry.Pro-inflammatory cytokine expression was determined using reverse transcription quantitative polymerase chain reaction.Results:TGW significantly ameliorated the severity of IMQ-induced psoriasis-like mouse skin lesions and restrained the activation of CD45^(+)cells,neutrophils and T lymphocytes(all P<0.01).Moreover,TGW significantly attenuated keratinocytes(KCs) proliferation and downregulated the mRNA levels of inflammatory cytokines including interleukin(IL)-17A,IL-23,tumor necrosis factor α,and chemokine(C-X-C motif) ligand 1(P<0.01 or P<0.05).Furthermore,it reduced the number of γδ T17 cells in skin lesion of mice and draining lymph nodes(P<0.01).Conclusions:TGW improved psoriasis-like inflammation by inhibiting KCs proliferation,as well as the associated immune cells and cytokine expression.It inhibited IL-17 secretion from γδ T cells,which improved the immune-inflammatory microenvironment of psoriasis.

Anti-inflammatory sesquiterpene polyol esters from the stem and branch of Tripterygium wilfordii

The stem and branch extract of Tripterygium wilfordii(Celastraceae)afforded seven new dihydroagarofuran sesquiterpene polyesters[tripterysines A‒G(1‒7)]and eight known ones(8‒15).The chemical structures of these new compounds were established based on combinational analysis of HR-ESI-MS and NMR techniques.The absolute configurations of tripterysines A‒C(1‒3)and E‒G(5‒7)were determined by X-ray crystallographic analysis and circular dichroism spectra.All the compounds were screened for their inhibitory effect on inflammation through determining their inhibitory effect on nitric oxide production in LPS-induced RAW 264.7 cells and the secretion of inflammatory cytokines TNF-αand IL-6 in LPS-induced BV2 macrophages.Compound 9 exhibited significant inhibitory activity on NO production with an IC50 value of 8.77μmol·L−1.Moreover,compound 7 showed the strongest inhibitory effect with the secretion of IL-6 at 27.36%.

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.

Inhibitory effects of Tripterygium wilfordii multiglycoside on benign prostatic hyperplasia in rats

The present study was designed to evaluate the inhibitory effects of Tripterygium wilfordii multiglycoside(GTW) against testosterone-induced benign prostatic hyperplasia(BPH) in rats. A total of 45 rats were randomly divided into five groups: Group I, vehicle control group(sham-operated and treated with vehicle); Group II, BPH group; Group III, BPH rats treated with finasteride at a dose of 5 mg·kg-1; and Groups IV and V, BPH rats treated with GTW at dose levels of 10 and 20 mg·kg-1, respectively. The drugs were administered orally once a day for 14 days. Prostate weight, prostatic index, and the testosterone and dihydrotestosterone(DHT) levels in serum and prostate, and the serum prostate specific antigen(PSA) levels were measured; prostate tissues were taken for histopathological examination; and serum biochemical analysis was also performed. The BPH rats displayed an increase in prostate weight, prostatic index with increased testosterone and DHT levels in both the serum and prostate, and increased serum PSA levels. GTW treatment at both doses resulted in significant reductions in prostate weight, prostatic index, testosterone and DHT levels in both the serum and prostate, and serum PSA levels, compared with BPH group. Histopathological examination also indicated that GTW treatment at both doses inhibited testosterone-induced prostatic hyperplasia. Serum biochemical analysis showed that the liver and renal functions were normal. In conclusion, GTW inhibited testosterone-induced prostatic hyperplasia in rats, without host toxicity, providing a basis for the development of GTW as a novel therapy for BPH.

Discovery of β-Dihydroagarofuran-Type Sesquiterpenoids from the Leaves of Tripterygium wilfordii with Neuroprotective Activities

Nine undescribed dihydro-β-agarofuran sesquiterpenoid derivatives(1—9),along with a known analogue(10),were obtained from the leaves of Tripterygium wilfordii.Their gross structures were determined via extensive spectroscopic data,and the absolute configurations were elucidated by means of single-crystal X-ray diffraction analysis and electron circular dichroism(ECD)techniques,which include ECD exciton chirality,octant rule of saturated cyclohexanone and comparison between the experimental and calculated ECD spectra.All the isolated compounds were tested for their neuroprotective activities against H2O2-induced cell injury in human neuroblastoma SH-SY5Y cells.Compounds 5 and 6 improved cell viability by 16.15%and 15.12%compared with the H2O2 treated group at 25μmol·L^(-1),respectively.

Molecular cloning and functional identification of a cDNA encoding 4-hydroxy-3-methylbut-2-enyl diphosphate reductase from Tripterygium wilfordii

The 4-hydroxy-3-methylbut-2-enyl diphosphate reductase(HDR) is the last step key enzyme of the methylerythritol phosphate(MEP) pathway,synthesizing isopentenyl diphosphate and its allyl isomer dimethylallyl diphosphate,which is important for regulation of isoprenoid biosynthesis.Here the full-length cDNA of HDR,designated TwHDR(GenBank Accession No.KJ933412.1),was isolated from Tripterygium wilfordii for the first time.TwHDR has an open reading frame(ORF) of 1386 bp encoding461 amino acids.TwHDR exhibits high homology with HDRs of other plants,with an N-terminal conserved domain and three conserved cysteine residues.TwHDR cDNA was cloned into an expression vector and transformed into an Escherichia coli hdr mutant.Since loss-of-function E.coli hdr mutant is lethal,the result showed that transformation of TwHDR cDNA rescued the E.coli hdr mutant.This complementation assay suggests that the TwHDR cDNA encodes a functional HDR enzyme.The expression of TwHDR was induced by methyl-jasmonate(MJ) in T.wilfordii suspension cells.The expression of TwHDR reached the highest level after 1 h of MJ treatment.These results indicate that we have identified a functional TwHDR enzyme,which may play a pivotal role in the biosynthesis of diterpenoid triptolide in T.wilfordii.

Tripterygium wilfordii multiglycoside-induced hepatotoxicity via inflammation and apoptosis in zebrafish

Tripterygium wilfordii multiglycoside(GTW)is a commonly used compound for the treatment of rheumatoid arthritis(RA)and immune diseases in clinical practice.However,it can induce liver injury and the mechanism of hepatotoxicity is still not clear.This study was designed to investigate GTW-induced hepatotoxicity in zebrafish larvae and explore the mechanism involved.The 72 hpf(hours post fertilization)zebrafish larvae were administered with different concentrations of GTW for three days and their mortality,malformation rate,morphological changes in the liver,transaminase levels,and histopathological changes in the liver of zebrafish larvae were detected.The reverse transcription-polymerase chain reaction(RT-PCR)was used to examine the levels of microRNA-122(miR-122)and genes related to inflammation,apoptosis,cell proliferation and liver function.The results showed that GTW increased the mortality of zebrafish larvae,while significant malformations and liver damage occurred.The main manifestations were elevated levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST),significant liver atrophy,vacuoles in liver tissue,sparse cytoplasm,and unclear hepatocyte contours.RT-PCR results showed that the expression of miR-122 significantly decreased by GTW;the mRNA levels of inflammation-related genes il1β,il6,tnfα,il10,cox2 and ptges significantly increased;the mRNA level of tgfβsignificantly decreased;the mRNA levels of apoptosis-related genes,caspase-8 and caspase-9,significantly increased;the mRNA level of bcl2 significantly decreased;the mRNA levels of cell proliferation-related genes,top2αand uhrf1,significantly reduced;the mRNA levels of liver function-related genes,alr and cyp3c1,significantly increased;and the mRNA level of cyp3a65 significantly decreased.In zebrafish,GTW can cause increased inflammation,enhanced apoptosis,decreased cell proliferation,and abnormal expression of liver function-related genes,leading to abnormal liver structure and function and resulting in hepatotoxicity.