Objective To explore the effect of tri pterygium glycoside (TG) on the skeletal muscle atrophy and apoptosis after nerve allograft. Methods Twenty Wistar male rats were adopted as donors, weighing 200-250 g, and the sciatic nerves were harvested. Fifty SD male rats were adopted as recipients, weighing 200-250 g. Fifty SD rats were made the models of10 mm right sciatic nerve defect randomly divided into five groups (n=10): group A, group B, group C, group D and group E.groups A and B received fresh nerve allograft, groups C and D received sciatic nerve allograft pretreated with TG, and group E received autograft. The SD rats were given medicine for 5 weeks from the second day after the transplantation: groups A and E were given physiological sal ine, groups B and D TG 5 mg/ (kg·d), and group C TG 2.5 mg/ (kg·d). At 3 and 6 weeks, respectively, after nerve transplantation, general observation was performed; the structure of skeletal muscles was observed by HE staining; the diameter of skeletal muscles was analyzed with Image-Pro Plus v5.2; the ultrastructure of skeletal muscles was observed by TEM; the expressions of Bax and Bcl-2 were detected by immunohistochemical staining; and the apoptosis of skeletal muscles was detected by TUNEL. Results All rats survived to the end of the experiment. In general observation, the skeletal muscles of SD rates atrophied to different degrees 3 weeks after operation. The muscular atrophy in group A was more serious at 6 weeks, and that in the other groups improved. The wet weight, fiber diameter and expression of Bcl-2 in group A were significantly lower than those in groups B, C, D and E (P lt; 0.01)；those in groups B, C and D were lower than those in group E (P lt; 0.05); and there were no significant differences among groups B, C and D (P gt; 0.05). The apoptosis index and expression of Bax in group A were significantly higher than those in groups B, C, D and E (P lt; 0.01)；those in groups B, C and D were higher than in groupE (Plt; 0.05); and there were no significant differences among groups B, C and D (P gt; 0.05). Three weeks after nerve allograft, under the l ight microscope, the muscle fibers became thin; under the TEM, the sarcoplasmic reticulum was expanded. Six weeks after nerve allograft, under the l ight microscope, the gap of the muscle fibers in group A was found to broaden and connective tissue hyperplasia occurred obviously; under the TEM, sarcomere damage, serious silk dissolution and fragmentary Z l ines were seen in group A, but the myofibrils were arranged tidily in the other groups, and the l ight band, dark band and sarcomere were clear. Conclusion TG can decrease the skeletal muscle atrophy and apoptosis after nerve allograft. The donor’s nerve that is pretreated with TG can reduce the dosage of immunosuppressant for the recipient after allograft.
Objective To investigate the anti-rejection effect and the mechanism of triptolide （TPT） on islet allo- grafts in a murine model. Methods BALB/c mice were used as islet donor. C57BL/6 mice were rendered diabetic by streptozotocin （STZ） injection， and transplanted with islets under the left kidney capsule. The recipients were randomly （method of random digits table） divided into three groups （n=8）. The mice in the treatment groups were injected intrap-eritoneally with TPT at 50 μg/kg （low-dose TPT group， L-TPT group） or 100 μg/kg （high-dose TPT group， H-TPT group） daily in the first 5 days and then on alternate days until 14 days；while the mice in control group were given vehicles （1% tween 80）. Blood glucose after operation were monitored. The grafts were defined as rejection when two consecutive reading of blood glucose＞20 mmol/L. The left kidney of three recipients in each group were resected for pathological examination. The proportion of CD4＋CD25＋Foxp3＋ regulatory T cells in spleen tissues were tested by flow cytometry. Results The median survival time of islet allografts from the control group， L-TPT group， and H-TPT group were 12.6 days （9-16 days）， 21.4 days （14-27 days） ， and 27.6 days （19-34 days）， respectivly. The percentageof CD4＋CD25＋Foxp3＋regulatory T cells in spleen tissues of three groups were （5.2±0.6）%， （12.0±1.3）%， and（15.7±1.8）%， respectivly. Compared with control group， the median survival time of islet transplantation in mice exte-nded and the proportion of CD4＋CD25＋Foxp3＋ regulatory T cells in spleen tissues increased （P＜0.05）. Conclusions TPT could increase the percentage of CD4＋CD25＋Foxp3＋ regulatory T cells， reduce the rejection after islet transplanta-tion， and prolong the survival time of islet transplantation in mice. The immunosuppressive effect of TPT shows a dose-dependent.
ObjectiveTo investigate the effects of celastrol on the growth and apoptosis of huamn hepatoma SMMC-7721 cells, and investigate its preliminary action mechansim.
MethodsSMMC-7721 cells were cultured in vitro, CCK-8 assay and Annexin V-FITC/PI staining method were conducted to investigate the effects of celastrol on the growth and apoptosis of huamn hepatoma SMMC-7721 cells after the cells were treated with drugs, and then the Caspase-3 activity and NF-κB protein expression were determined by Caspase-3 activity determination kit and Western blot. Huamn hepatoma SMMC-7721 cells transplantation tumor models in nude mice were established and the effect of celastrol on the growth of transplantation tumor were observed.
ResultsCelastrol could inhibit the SMMC-7721 cells growth in a dose and time dependent manner. Annexin-V/PI staining showed that SMMC-7721 cells were induced to death with the concentration increasing of celastrol. Caspase-3 activity was measured after treatment with celastrol and the results indicated that the activity of caspase-3 was significantly enhanced. Western blot experiments showed that the expression of NF-κB protein decreased in a time-dependent manner after treatment with celastrol. Celastrol could inhibit SMMC-7721 cells transplantation tumor growth in nude mice.
ConclusionsCelastrol could inhibit the proliferation of human hepatoma SMMC-7721 cells and induces apoptosis, and inhibit SMMC-7721 cell transplantation tumor growth in nude mice. Celastrol induce apoptosis of SMMC-7721 cells might through activating Caspase-3 pathway and NF-κB pathway.
Objective To evaluate the efficacy and safety of tripterygium for diabetic nephropathy. Methods All randomized or quasi-randomized controlled trials (RCTs or quasi-RCTs) of tripterygium for biabetic nephropathy were collected from The Cochrane Library (Issue 1, 2010), MEDLINE (1996 to March 2010), CNKI (1994 to March 2010), and CBM (1978 to March 2010). Two reviewers evaluated the quality of the trials and extracted the data independently. RevMan 5.0 software was used for meta-analyses. Results A total of 12 RCTs involving 862 patients were identified. The methodology of the included trials was poor and potential publication bias existed. The meta-analyses results showed: (1) Compared with the conventional treatment, the tripterygium showed more effects in reducing the 24-hour urinary protein (Clinical phase: WMD= –0.49, 95%CI –0.63 to –0.34, No phase: WMD= –0.60, 95%CI –0.96 to –0.24), and the urinary albumin excretion rate (UAER) (WMD= –148.75, 95%CI –238.01 to –59.48) was higher than that of the conventional treatment. (2) There were no significant differences between the two groups in the effect on the serum creatinine (Clinical phase: WMD= –8.43, 95%CI –18.15 to 1.29, No phase: WMD= –0.66, 95%CI –2.12 to 0.79) and creatinine clearance rate (WMD= 1.74, 95%CI –6.34 to 9.83). (3) Without enough data, it was uncertain to define the effect of tripterygium on lipids, blood pressure of the DN patients. (4) No severe adverse events or allergic reactions were reported. Conclusion Tripterygium may be a kind of medicine relatively safe and effective for diabetic nephropathy. However, the evidence is not b enough because of some low-quality trials and publication bias. Rigorously-designed, randomized, double-blind, and placebo-controlled trials of tripterygium for diabetic nephropathy are needed to further assess the effect.
Objective Tri ptol ide can suppress immunological rejection reaction. To investigate the effect of tri ptol ide on allogenic tendon transplantation in repairing tendon defect in chicken. Methods The defect model of the third toes tendon was establ ished in 64 healthy-cleaning male Leghorn chickens (4-month-old, weighing 1.9-2.3 kg), which underwent allogenic tendon transplantation for repairing and were divided into 2 groups randomly (n=32). Tri ptol ide feeding[100 μg/(kg·d)] was given for 3 weeks in the experimental group and normal feeding in the control group. General condition of the chickens was observed after operation. The transplanted tendons were harvested from 4 chickens in each group for gross observation at 1, 2, 3, and 4 weeks after operation; the histological observation was performed at 1 and 3 weeks, and transmission electron microscope observation at 2 and 4 weeks. The blood and tendon were harvested from another 8 chickens in each group for flow cytometry and biomechanical tests respectively at 3 and 6 weeks. Results All chickens survived to the experiment end. Gross observation: with time extending, hyperemia and edema around transplanted tendon were rel ieved. Rarefaction adhering zone was seen in experimental group, and pyknotic adhering zone in control group. Histological observation: inflammatory reaction in experimental group was sl ighter than that in control group at 1 and 3 weeks. Transmission electron microscope observation: at 2 and 4 weeks, fibroblasts had big cell nucleus, more euchromatin, and l ittle heterochromatin in experimental group; however, there were small amount of rough endocytoplasmic reticulums with gentle expanded capsular space in control group, which contained sparse content. Flow cytometry test: at 3 and 6 weeks, peri pheral blood contained less CD4+ and CD8+ T lymphocytes in experimental group than in control group, and the ratio of CD4+ to CD8+ T lymphocyte significantly decreased in experimental group when compared with control group (P lt; 0.05). Biomechanical examination: at 3and 6 weeks, the maximum tensile strength in experimental group was bigger than that in control group, and tensile adhesion power in experimental group was smaller than that in control group. There were significant differences in the indexes between 2 groups (P lt; 0.05). Conclusion Tri ptol ide can suppress immunological rejection reaction, strengthen tendon healing strength, and reduce tendon adhesion in allogenic tendon transplantation.
Objective To investigate the appropriate concentration of tripterygium wilfordii and immunological rejection of rats’ sciatic nerve allograft with the tripterygium wilfordii’s pretreatment so as to explore tripterygium wilfordii’ s suppression. Methods Sixty SD rats (male, weighing 270-290 g), as sciatic nerve allograft acceptor were randomized into5 groups (groups A, B, C, D and E, n=12). To repair the sciatic nerve defect of SD rats, the Wistar rats’ sciatic nerve allografts about 15 mm long were used with 24 hours’ soak of different concentrations of tripterygium wilfordii (group A: 200 mg/L, group B: 400 mg/L, group C: 800 mg/L). The control groups (group D: the fresh sciatic nerve allograft from donors; group E: the fresh sciatic nerve allograft from themselves) were establ ished. At different time points after operation, the morphological examinations (the observation of histology, l ight microscope, electron microscope), the detection of myelin basic protein’s (MBP) content and the analyses of CD4+ and CD8+ T cells on the allografts in the acute phase were performed Results There was no significant difference in morphology among groups A, B and C, the adhesions between allografts and connective tissue were milder than that of group D, and the allografts’ morphous and the inflammatory cell infiltration were better than that of group D. The degeneration of myel in sheath was observed at different levels and there was no significant difference between group B and group E (P gt; 0.05). There was a significant difference in immunological rejection between groups A, B, C and group D (P lt; 0.05). Conclusion Tripterygium wilfordii can effectively suppress the acute immunological rejection in the early stage after operation, and protect the myel in sheath to a certain extent.
