TRAIL通过内质网应激对肝星状细胞凋亡的调控及部分机制的研究  

Effects of TRAIL on the hepatic stellate cells apoptosis in hepatic stellate cells and relevant mechanism

作　　者：李晓慧[1,2] 李俊[1,2] 黄艳[1,2] 黄成[1,2] 王雅蕊 王欢[1,2] 孙仕伟 黄蒙蒙[1,2] 

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学肝病研究所,合肥230032

出　　处：《安徽医科大学学报》2014年第7期863-867,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81072686;81273526;81102493);高等学校博士学科点专项科研基金(编号:20103420120001);安徽医科大学2013年国家级大学生创新创业训练计划(编号:201310366030)

摘　　要：目的探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)与内质网应激(ERS)相关蛋白对大鼠肝星状细胞系(HSC-T6)凋亡的影响及内在联系。方法体外血小板源性生长因子(PDGF)诱导活化的肝星状细胞(HSC),再分别加入TRAIL与ERS诱导剂衣霉素(TN)处理后,Hoechst33258染色和流式细胞仪检测细胞凋亡变化;RT-PCR法检测mRNA的表达;Western blot检测蛋白表达变化。结果与PDGF诱导活化的HSC组作为对照组比较,荧光显微镜观察到TRAIL与TN分别处理的HSC,Hoechst 33258染色细胞呈核固缩等凋亡形态变化,同流式细胞仪检测结果;与对照组相比,RT-PCR和Western blot结果显示,TRAIL组与TN组细胞凋亡基因Bax mRNA、凋亡蛋白C-Caspase-3的表达明显上调,而且ERS蛋白C-Caspase-12、CHOP、GRP78的表达均明显上调。结论 TRAIL能通过明显上调凋亡基因Bax mRNA、凋亡蛋白C-Caspase-3的表达,诱导PDGF活化的HSC凋亡,其机制可能与诱导ERS相关蛋白CHOP、GRP78、Caspase-12的表达有关。Objective To detect the effect of ER stress and TRAIL in rat hepatic stellate cell lines （ HSC-T6 ） and to explore the possible apoptosis pathway. Methods Hepatic stellate cell （ HSC） apoptosis was measured by Hoechst staining apoptosis detection kit and flow cytometric analysis. CHOP,GRP78,C-Caspase-12 and-3 protein expression were analyzed by Western blot;Bax mRNA expression was evaluated by RT-PCR. Results It showed that PDGF-induced HSC as control. The increased of HSC apoptosis was measured by Hoechst 33258 and flow cytometry. In addition, TRAIL increased the expression of Bax and C-Caspase-3 in activated HSC. TRAIL also increased C-Caspase-12,CHOP and GRP78 in activated HSC. These results suggested that TRAIL induced apoptosis through ERS-mediated pathways in HSC. Conclusion These results demonstrate that TRAIL induces apoptosis through ERS-mediated pathways in HSC.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 肝星状细胞 内质网应激 凋亡 CASPASE-12 

分 类 号：R575.2[医药卫生—消化系统]