Sec61α 在肝细胞脂肪变性模型中的动态变化及可能作用机制.docVIP

Sec61α在肝细胞脂肪变性模型中的动态变化及可能作用机制 王 军1，黄殿静2，赖姝婕1，熊 吉1，魏艳玲1，李 青1，李 砚1，陈潇迪1，王 斌1，陈东风1 (400042 重庆，第三军医大学大坪医院野战外科研究所消化内科1；210024，南京，武警江苏总队第二支队卫生队2) [摘要] 目的 探讨Sec61α在肝细胞脂肪变性模型中的动态变化及可能作用机制。方法 应用软脂酸与油酸混合物诱导L02肝细胞发生脂肪变性，建立非酒精性脂肪性肝病肝细胞体外模型，按0、2、4、8h及16h收获细胞；随后应用Sec61抑制剂Eeyarestatin 1处理肝细胞，分为正常对照组、脂肪变性组、DMSO组及抑制剂组，通过油红O染色、qRT-PCR及Western blot检测Sec61α、GRP78、GRP94、Bcl-2及Bax的表达变化。结果 软脂酸与油酸混合物成功诱导肝细胞发生脂肪变性。与对照细胞相比，脂肪变肝细胞Sec61αmRNA相对表达量在脂肪变早期下降（2、4h，P0.05)，晚期升高（8、12 、16h，P0.05)；GRP78在8h开始升高，16h达到高峰（P＜0.01）。Sec61α与GRP78的蛋白水平与mRNA水平表达在时间梯度上变化一致。Eeyarestatin 1处理显著减轻肝细胞脂肪变程度；与脂肪变性组相比，抑制剂组肝细胞Sec61α、GRP78与Bax明显降低（P＜0.01），GRP94、 Bcl-2则显著升高（P＜0.01）。结论 Sec61α可能通过调节肝细胞内质网应激，参与肝细胞脂肪变的形成。 [关键词] 脂肪变性；内质网应激；Sec61α；GRP78；GRP94；Bcl-2；Bax [中图法分类号] R575.5；R329.25；R363.21； [文献标志码] A Dynamic expression of Sec61 alpha and its possible mechanisms in hepatic steatosis model Wang Jun1, Huang Dianjing2, Lai Shujie1, Xiong Ji1, Wei Yanling1, Li Qing1, Li Yan1, Chen Xiaodi1, Wang Bin1, Chen Dongfeng1 (1Department of Gastroenterology, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042; 2Health Team, Detachment of the Second, Armed Police Jiangsu Corps, Nanjing, 210036, China) ［Abstract］Objective To examine the expression change of Sec61α in hepatic steatosis and its possible mechanism. Methods L02 liver cells were treated with or without palmitic acid and oleic acid mixture to establish an in vitro cellular model of NAFLD (control group or steatosis group). The steatotic hepatocytes were further incubated with or without a Sec61 inhibitor Eeyarestatin 1 (DMSO group or inhibitor group). The cells were collected at 0, 2, 4, 8, 12, 16h after induction for, oil red O staining, qRT-PCR, or Western blot analysis to detect the expression of Sec61 alpha, GRP78, GRP94, Bcl-2, and Bax. Results Liver cells steatosis was induced by palmitic acid and oleic acid mixture. As compared to control cells, steatotic hepatocytes expressed decre