慢病毒介导ACE2过表达对血管紧张素Ⅱ诱导的大鼠肝细胞的白蛋白表达及迁移能力的影响  

Lentivirus-mediated angiotensin-converting enzyme 2 overexpression inhibits angiotensin Ⅱ-induced albumin down-regulation and enhancement of migration in rat hepatocytes

作　　者：张莉莉[1] 张文雍[1] 李洋[1] 李旭[2] 

机构地区：[1]南方医科大学南方医院急诊科,广东广州510515 [2]南方医科大学南方医院感染内科,广东广州510515

出　　处：《南方医科大学学报》2015年第5期652-658,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(81070338);中国肝炎防治基金会王宝恩肝纤维化研究基金(20090018);广东省科技计划项目2010B060500008)~~

摘　　要：目的探讨血管紧张素转换酶2(Angiotensin-converting enzyme 2,ACE2)过表达对血管紧张素Ⅱ(Angiotensin II,AngⅡ)诱导的肝细胞白蛋白表达下调及细胞迁移性增强的抑制作用。方法常规培养大鼠肝细胞系BRL-3A细胞株,予以Ang II(10-7mol/L)进行刺激,观察不同处理时间(24、48 h)组肝细胞内白蛋白、波形蛋白(vimentin)表达及细胞迁移性的改变;通过构建lenti ACE2慢病毒载体,建立稳定过表达ACE2的细胞系,并分别用AngⅡ、A779进行干预,观察肝细胞内相应蛋白表达及其迁移性的改变。采用细胞免疫荧光、Western blot检测细胞中蛋白水平变化;Transwell迁移实验观察不同处理组细胞迁移能力的改变。结果 AngⅡ处理组肝细胞内vimentin表达显著增加、albumin表达减低,细胞迁移能力显著增强,并具有时间依赖效应。ACE2过表达大鼠肝细胞albumin表达明显升高,vimentin蛋白表达下降,该作用被MAS受体抑制剂A779阻断。同时ACE2过表达显著抑制AngⅡ的作用,vimentin合成显著下降,albumin表达水平显著升高,细胞迁移能力受到抑制。结论 ACE2过表达可抑制AngⅡ诱导的肝细胞albumin表达下调及迁移能力增强。Objective To investigate the inhibitory effect of angiotensin-converting enzyme 2 （ACE2） on angiotensin II （Ang II）-induced down-regulation of albumin expression and enhancement of cell migration in rat hepatocytes. Methods Cultured rat hepatocyte were treated with Ang II （10- 7 mol/L） for different time lengths, and the protein expressions of vimentin and albumin and cell migration were detected. The cells transfected with lentiGFP or lentiACE2 were treated with A779 for 1 h and then with Ang II, and Western blotting and immunofluorescent cytochemistry were used to detect the protein levels;the cell migration was evaluated by Transwell assay. Result Ang II induced significantly increased vimentin expression and reduced albumin expression in rat hepatocytes in a time-dependent manner. Overexpression of ACE2 obviously inhibited the up-regulation of vimentin expression, reduction of albumin expression, and enhancement of cell migration induced by Ang II. Conclusion ACE2 overexpression can inhibit Ang II-induced up-regulation of vimentin, reduction of albumin expression, and enhancement of cell migration in rat hepatocytes.

关 键 词：血管紧张素转换酶2 血管紧张素Ⅱ ALBUMIN VIMENTIN 迁移 肝细胞 肝纤维化 

分 类 号：R965[医药卫生—药理学]