转录因子Twist促进腹膜透析患者腹膜纤维化的作用机制研究  

Mechanism of transcription factor Twist on promoting peritoneal fibrosis in patients undergoing peritoneal dialysis

作　　者：贾真[1,2] 姬丽华[1] 陈景荷[1] 梁维[1] 车明文 何丽洁[1] 孙世仁[1] 王汉民[1] 

机构地区：[1]第四军医大学西京医院肾脏内科,西安710032 [2]西安市第一医院肾脏内科

出　　处：《解放军医学杂志》2013年第11期879-884,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81270768;81270849);陕西省科技攻关项目(2012K16-08-05)~~

摘　　要：目的探讨转录因子Twist对腹膜透析患者腹透流出液人腹膜间皮细胞(HPMCs)的作用及相关机制。方法将腹膜透析患者透出液离心后进行HPMCs培养,检测Twist、E-cadherin、α-SMA、Bmi-1的蛋白及免疫荧光表达。分别采用上调质粒pcDNA3.1-Twist和空载体pcDNA3.1转染HPMCs,检测Twist、E-cadherin、α-SMA以及Bmi-1的蛋白及免疫荧光表达。分别采用Twist的siRNA质粒和空载体pSlience转染转分化的HPMCs,检测Twist、E-cadherin、α-SMA以及Bmi-1的蛋白及免疫荧光表达。采用Bmi-1的siRNA质粒转染转分化的HPMCs,检测E-cadherin、α-SMA以及Bmi-1的蛋白及免疫荧光表达。结果免疫荧光及Western blotting检测结果显示,随着透龄增加,E-cadherin表达降低,Twist、α-SMA及Bmi-1表达增加(P<0.05)。与空载体组相比,HPMCs过表达Twist后,E-cadherin表达减弱,Twist、α-SMA及Bmi-1表达增加(P<0.05)。用小干扰RNA使Twist沉默后,E-cadherin表达增加,Twist、α-SMA及Bmi-1表达减弱(P<0.05)。用小干扰RNA使Bmi-1沉默后,E-cadherin表达增加,α-SMA、Bmi-1表达减弱(P<0.05)。结论 Twist参与了腹膜纤维化的发生发展过程,其作用部分是通过Bmi-1促进HPMCs的转分化实现的。Objective To explore the mechanism of transcription factor Twist on promoting peritoneal fibrosis of human peritoneal mesothelial cells （HPMCs） in patients undergoing peritoneal dialysis （PD）. Methods The HPMCs in peritoneal fluid after dialysis from patients underwent peritoneal dialysis was collected and cultured, the expression and subcellular localization of E-cadherin, α-SMA, Twist and Bmi-1 were assayed by immunofluorescence and Western blotting. The expression of Twist, Bmi-1, E-cadherin and α-SMA were determined by immunofluorescence and Western blotting after the plasmids pcDNA3.1-twist and empty vector pcDNA3.1 were transfected into HPMCs with Lipofectamine 2000 in vitro. Twist, Bmi-1, E-cadherin and α-SMA were determined by Western blotting and immunofluorescence after silencing the expression of Twist by small interfering RNA （siRNA） and empty vector pSlience. Bmi-1, E-cadherin and α-SMA were assessed by Western blotting and immunofluorescence after silencing the expression of Bmi-1 by siRNA. Results The expression of E-cadherin decreased gradually and the expression of Twist, α-SMA and Bmi-1 increased along with the increase in PD time as determined with immunofluorescence and Western blotting. Compared with that expressed in the HPMCs trasfected with empty vector pcDNA3.1, the expression of E-cadherin significantly reduced and the expressions of α-SMA, Twist and Bmi-1 increased significantly after transfection of the plasmids pcDNA3.1-twist （P〈0.05）. The expression of E-cadherin increased significantly and the expressions of α-SMA, Twist and Bmi-1 decreased significantly after silencing the expression of twist by siRNA. The expression of E-cadherin increased significantly and the expressions of α-SMA and Bmi-1 decreased significantly after silencing the expression Bmi- 1 by siRNA. Conclusion Twist may be involved in the process of peritoneal fibrosis, and its underlying mechanism is the promotion of the transdifferentiation of HPMCs via the regulation of Bmi-1.

关 键 词：腹膜纤维化 上皮-间质细胞转分化 TWIST转录因子 BMI-1 钙黏着糖蛋白类 肌动蛋白类 

分 类 号：R459.5[医药卫生—治疗学]