Molecular mechanism of apoptosis in rat hepatic stellate cells HSC-T6 induced by TRAIL
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摘要: 目的研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导肝星状细胞凋亡情况及调控机制。方法用RT-PCR检测大鼠肝星状细胞(HSC-T6)中α-SMAmRNA和DR5mRNA的表达;MTT比色法和流式细胞术检测外源性TRAIL对HSC-T6细胞增殖和诱导细胞凋亡的影响;采用Western blot检测Bax、Caspase3蛋白的表达。结果培养的HSC-T6细胞表达α-SMA mRNA和DR5mRNA随作用时间的延长逐渐增加,TRAIL可以抑制其细胞增殖。TRAIL与对照组比较可以诱导活化的HSC-T6细胞凋亡明显增加(P<0.05),Western-blotting分析显示TRAIL作用下,HSC-T6细胞中的线粒体Bax蛋白、细胞浆Caspase3蛋白表达均上调。结论外源性TRAIL可诱导活化的HSC-T6细胞凋亡,可能与DR5及线粒体Bax表达上调有关。Abstract: Objective To investigate the effect and mechanism of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on apoptosis in cultured rat hepatic stellate cells (HSCs) HSC-T6. Methods The gene expressions of α-SMA and DR5 in HSC-T6cells were measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) .The proliferation and apoptosis of HSC-T6 cells treated with TRAIL were detected by 3- (4, 5-dimethytthiazole-2-yl) -2, 5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. The Bax, Caspase3 in the HSC-T6 cell s homogenates and mitochondrion were detected by western blot. Results The gene expressions of α-SMA and DR5 in cultured rat HSC-T6cells was low in the 2nd and 4th days, increased significantly in the 8th and 12th days (P<0.05) . The proliferation of HSC-T6 cells was reduced by TRAIL.TRAIL also induced apoptosis of HSC-T6 cell s dramatically than control (P<0.05) .The mitochondrion bax and cleaved active subunits of caspase-3 were highly expressed in treated HSC-T6 cells than in the control. Conclusion TRAIL can induce apoptosis of HSC-T6 cells effectively and the high expression of DR5 and Bax may play an important role in the apoptosis.
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Key words:
- tumor necrosis factor-alpha /
- apoptosis /
- astrocytes /
- mitochondria
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