纤连蛋白对肿瘤坏死因子相关凋亡诱导配体诱导肝星状细胞凋亡的影响

Effect of fibronectin on protection of rat hepatic stellate cells HSC-T6 from TRAIL-induced apoptosis

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。

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 HSCT6 cells was detected by 3 - （4, 5 -dimethytthiazole -2 -yl） -2, 5 - diphenyltetrazolium bromide （MTY） assay and apoptosis rate was detected by flow Cytometry. Western Blot analysis was performed to de- termine the FAK ,pFAK ,Ban in the HSC - T6 cells bomogenates 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 expres- sions of DFAK and low level expression Bax may olav an important role.