Effect of fibronectin on protection of rat hepatic stellate cells HSC-T6 from TRAIL-induced apoptosis
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摘要: 目的研究纤连蛋白(FN)对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导肝星状细胞(HSC)凋亡的影响及调控机制。方法 MTT比色法、流式细胞术检测加入FN时,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检测粘着斑激酶(FAK)、磷酸化粘着斑激酶(pFAK)、Bax的表达。结果 TRAIL可以抑制HSC-T6细胞增殖,可以诱导HSC-T6细胞凋亡,而FN的存在可以使TRAIL诱导的HSC-T6细胞凋亡明显减少(P<0.05),Western Blot分析显示TRAIL诱导HSC-T6细胞凋亡时加入FN,细胞浆pFAK表达上调而线粒体Bax表达下降。结论 FN可以使外源性TRAIL诱导的HSC-T6细胞凋亡减少,机制可能与FN促使FAK磷酸化,pFAK表达上调并减少线粒体Bax表达有关。Abstract: Objective To determine whether fibronectin can protect rat hepatic stellate cells (HSCs) HSC-T6 from TRAIL-induced apoptosis and to study the mechanism involved in this effect.Methods HSC-T6 were plated on fibronectin-uncoated or coated culture wells and treated with TRAIL.The inhibition on the proliferation of HSC-T6 cells was detected by 3- (4, 5-dimethytthiazole-2-yl) -2, 5-diphenyltetrazolium bromide (MTT) assay and apoptosis rate was detected by flow Cytometry.Western Blot analysis was performed to determine the FAK、pFAK、Bax in the HSC-T6 cells homogenates and mitochondrion.Results FN could increase cell proliferation of HSC-T6 cells treated with TRAIL aand reduce apoptosis rate dramatically (P<0.05) .Western Blot showed that FN increased pFAK expression and reduced Bax expression.Conclusion FN could reduce apoptosis of HSC-T6 cells treated with TRAIL effectively, the high expressions of pFAK and low level expression Bax may play an important role.
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Key words:
- hepatic stellate cell /
- fibronectins /
- tumor necrosis factor-alpha /
- ligands /
- apoptosis
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