Application of modified two-cuff technique and multiglycosides tripterygium wilfordii in hamster-to-rat liver xenotransplant model.
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AIM: To modify the hamster-to-rat liver xenotransplant technique to prevent postoperative complications, and to study the inhibiting effect of multiglycosides tripterygium wilfordii (T(II)) on immune rejection. METHODS: Female golden hamsters and inbred male Wistar rats were used as donors and recipients, respectively. One hundred and twelve orthotopic liver xenotransplants were performed by Kamada's cuff technique with modifications. Over 72 hour survival of the animal after operation was considered as a successful operation. When the established surgical model became stable, 30 of the latest 42 cases were divided into untreated control group (n=15) and T(II) group (n=15) at random. Survival of recipients was observed. Liver specimens were collected at 2 and 72 h from the operated animals and postmortem, respectively, for histological study. RESULTS: The successfully operative rate of the 30 operations was 80 %, and the survival of the control and T(II) group was 7.1+/-0.35 was days and 7.2+/-0.52 days, respectively (t=0.087,P=0.931). The rate of conjunctival hyperemia in control group (100 %) differed significantly from that (31 %) in T(II) group (P=0.001). Rejection did not occur in both groups within 2 h postoperatively, but became obvious in control group at 72 h after surgery and mild in T(II) group. Although rejections were obvious in both groups at death of recipients, it was less severe in T(II) group than in control group. CONCLUSION: This modified Kamada's technique can be used to establish a stable hamster-to-rat liver xenotransplant model. Monotherapy with multiglycosides tripteryguiumwilfordii (30 mg x kg(-1) x d(-1)) suppresses the rejection mildly, but fails to prolong survival of recipients. (+info)
Analysis of triptolide-regulated gene expression in Jurkat cells by complementary DNA microarray.
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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. (+info)
Inhibition by triptolide of IL-1-induced collagen degradation by corneal fibroblasts.
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PURPOSE: Extracts of the herb Tripterygium wilfordii hook f, the major component of which is triptolide, have been used in traditional Chinese medicine for the treatment of rheumatoid arthritis. Triptolide also exerts many other biological actions both in vitro and in vivo. The effect of this agent on collagen degradation by cultured corneal fibroblasts was examined. METHODS: Rabbit corneal fibroblasts were cultured in three-dimensional gels of type I collagen and in the absence or presence of interleukin (IL)-1beta or triptolide. The extent of collagen degradation was determined by measurement of the amount of hydroxyproline generated by acid-heat hydrolysis of the culture supernatants. The activities of matrix metalloproteinase (MMP)-1 and plasmin were measured with the specific substrates thiopeptolide and S-2251, respectively. The release of MMPs into the culture supernatant was assessed by immunoblot analysis and gelatin zymography, and the abundance of MMP mRNAs in the cells was determined by reverse transcription and real-time polymerase chain reaction. RESULTS: Triptolide inhibited the IL-1beta-induced degradation of collagen by corneal fibroblasts in a dose- and time-dependent manner. Neither the activity of purified recombinant MMP-1 nor that of plasmin in culture supernatants was affected by triptolide. The IL-1beta-induced expression of MMP-1, -2, -3, and -9 by corneal fibroblasts was inhibited by triptolide at the protein or mRNA level. CONCLUSIONS: Triptolide inhibits collagen degradation by corneal fibroblasts by inducing downregulation of the production of MMPs, without directly affecting the collagenolytic activity of these enzymes. (+info)
Triptolide, an active compound identified in a traditional Chinese herb, induces apoptosis of rheumatoid synovial fibroblasts.
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BACKGROUND: Extracts of Tripterygium wilfordii Hook F (TWHF), a traditional Chinese herb, have been reported to show efficacy in patients with rheumatoid arthritis (RA). Since RA is not only characterized by inflammation but also by synovial proliferation in the joints, we examined whether triptolide (a constituent of TWHF) could influence the proliferation of rheumatoid synovial fibroblasts (RSF) by induction of apoptosis. RESULTS: RSF were obtained from RA patients during surgery and were treated with triptolide under various conditions. The viability and proliferation of RSF were measured by the 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) assay and by 5-bromo-2'-deoxyuridine incorporation, respectively. Apoptosis was identified by detection of DNA fragmentation using an enzyme-linked immunosorbent assay and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL). The role of caspases in apoptosis of RSF was analyzed by measuring caspase-3 activity. Activation of the peroxisome proliferator-activated receptor (PPAR) gamma was assessed by a luciferase reporter gene assay using RSF transfected with a plasmid containing the peroxisome proliferator response element. Triptolide decreased viability, inhibited proliferation, and induced apoptosis of RSF in a concentration-dependent manner at very low (nM) concentrations. Caspase-3 activity was increased by treatment with triptolide and was suppressed by caspase inhibitors. Although PPARgamma activation was induced by 15-deoxy-Delta12,14-prostaglandin J2, triptolide did not induce it under the same experimental conditions. An extract of TWHF also induced DNA fragmentation in RSF. CONCLUSION: The mechanism of action remains to be studied; however, triptolide may possibly have a disease-modifying effect in patients with RA. (+info)
Clinical study of qingluo tongbi granules in treating 63 patients with rheumatoid arthritis of the type of yin-deficiency and heat in collaterals.
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The study is to observe the therapeutic effects of qingluo tongbi granules (QTG) in patients with rheumatoid arthritis (RA) and the changes of immune indexes. In this series there are 63 patients with RA of the type of yin-deficiency and heat in collaterals treated with QTG as the treated group and 55 patients of the same type treated with Tripterygium glycosides as the control group. As a result, in the treated group, the curative rate is 9.52% and markedly effective rate 38.10%, with a total effective rate of 90.48%, while the corresponding rates in the control group are 0, 20.00% and 83.64%, respectively. The curative effect in the treated group is better than that in the control group (P<0.05). Besides, no obvious adverse reactions are found in the treated group. Therefore it is concluded that as a new medicinal preparation QTG is safe and effective in the treatment of RA. (+info)
Tripterygium hypoglaucum (level) Hutch induces aneuploidy of chromosome 8 in mouse bone marrow cells and sperm.
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Aneuploidy of mouse chromosome 8 induced by a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH) was investigated by fluorescence in situ hybridization (FISH) in vivo. Male mice were treated with THH (single i.p. injection) at doses of 120, 240 and 480 mg/kg. Colchicine (COL, 1.5 mg/kg i.p.) was used as a positive control. Bone marrow cells and epididymal sperm were collected 24 h and 22 days after treatment, respectively. Chromosome 8 aneuploidies induced by THH in bone marrow cells and sperm were determined by FISH with a biotin-16-dUTP labelled DNA probe corresponding to the centromeric region of chromosome 8. The hybridized probe was detected with avidin-FITC. The frequencies of trisomy 8 in bone marrow cells were 0.16% in the solvent control group, 0.39% in the COL-treated group and 0.33, 0.41 and 0.41% in the THH-treated groups, respectively. The frequencies of disomy 8 sperm were 0.11% in the solvent control group, 0.27% in the COL-treated group and 0.23, 0.27 and 0.27% in the THH-treated groups, respectively. The experiment showed that induced aneuploidy frequencies were higher in bone marrow cells than in sperm with COL and the two higher doses of THH (P < 0.05). All groups were significantly different from the corresponding solvent controls (P < 0.01-0.001), but there was no dose-related increase in either cell type. Considering the present results together with our previous studies, it appears that THH is a potent mammalian aneugen which may pose a genetic risk to human patients. (+info)
Triptolide, an active component of the Chinese herbal remedy Tripterygium wilfordii Hook F, inhibits production of nitric oxide by decreasing inducible nitric oxide synthase gene transcription.
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OBJECTIVE: The ethyl acetate (EA) extract of Tripterygium wilfordii Hook F (TWHF) and its major active component, triptolide, have been reported to be effective in the treatment of rheumatoid arthritis and other autoimmune inflammatory diseases. Nitric oxide (NO) has been recognized as an important mediator of inflammation. This study was therefore undertaken to examine the effects of the EA extract and triptolide on the production of NO and inducible NO synthase (iNOS) gene expression and transcription in vivo and in vitro. METHODS: Peritoneal macrophages from C57BL/6J mice treated orally with the EA extract of TWHF were assayed for NO production and iNOS messenger RNA (mRNA) expression by reverse transcriptase-polymerase chain reaction. The murine fibroblast cell line NIH3T3 was also assessed for NO production and iNOS mRNA expression, as well as for iNOS promoter activation, Oct-1 nuclear binding capacity, and Oct-1 protein content by transient transfection, electrophoretic mobility shift assay, and immunoblotting, respectively. RESULTS: NO production and iNOS mRNA expression by macrophages from C57BL/6J mice immunized with trinitrophenyl-bovine serum albumin in Freund's complete adjuvant were significantly inhibited by oral administration of the EA extract (52.3% and 59.8% of control, respectively, at one-eighth of the dose that is lethal for 50% of the animals [LD(50)] and 21.0% and 38.1% of control, respectively, at one-fourth the LD(50)). Moreover, the EA extract and triptolide significantly inhibited NO production in vitro in activated peritoneal macrophages, which reflected a decreased level of iNOS mRNA. Finally, triptolide inhibited promoter activity of the iNOS gene and induction of the activity of the regulator of iNOS transcription, Oct-1. CONCLUSION: The EA extract of TWHF and triptolide inhibit transcription of the iNOS gene. This may contribute to the antiinflammatory effects of this traditional herbal remedy. (+info)
Therapeutic effects of glucosides of Cheanomeles speciosa on collagen-induced arthritis in mice.
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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. (+info)