YAP修复TNF-α诱导的肠上皮细胞损伤  

YAP ameliorates TNF-α induced intestinal epithelial barrier damage

作　　者：纪莲[1] 叶晓琳[2] Ji Lian;Ye Xiaolin(Medical Research Center,Shengjing Hospital of China Medical University,Key Laboratory of Research and Application of Animal Models for Environmental and Metabolic Diseases,Shenyang 110004,China;Department of Pediatric Gastroenterology,Shengjing Hospital of China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院实验研究中心辽宁省环境与代谢疾病动物模型研究与应用重点实验室,沈阳110004 [2]中国医科大学附属盛京医院小儿消化科,沈阳110004

出　　处：《国际儿科学杂志》2020年第8期584-588,共5页International Journal of Pediatrics

基　　金：辽宁省科技厅博士科研启动基金计划项目(2020-BS-092)。

摘　　要：目的探讨Yes相关蛋白(Yes-associated protein,YAP)在肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导的肠黏膜屏障损伤中的保护作用及潜在的分子机制。方法培养人克隆结肠癌Caco-2细胞14 d体外建立肠黏膜屏障模型,随后转染YAP质粒和对照质粒,24 h后加入TNF-α刺激损伤肠黏膜屏障,48 h后通过细胞计数试剂盒(cell counting kit WST-8,CCK-8)检测肠上皮细胞增殖情况,流式细胞术检测细胞周期,迁移小室(Transwell)检测细胞迁移能力。结果与正常对照组相比,加入TNF-α刺激后,肠上皮细胞增殖活性明显下降,细胞周期G0/G1期比例增加,S期和G2/M期比例减少,而YAP过表达可抑制TNF-α引起的细胞增殖活性下降,使细胞周期顺利从G0/G1期向S期和G2/M期过渡。Transwell结果表明,正常对照组细胞迁移数目为(172.4±13.1)个细胞,加入TNF-α刺激后,细胞的迁移数目降至(80.4±9.3)个细胞,而预先转染YAP可抑制TNF-α引起的细胞迁移能力下降,细胞迁移数目为(124.2±9.2)个细胞,差异有统计学意义(P<0.01)。结论YAP通过促进肠上皮细胞增殖和迁移从而抑制TNF-α诱导的肠黏膜屏障损伤。Objective To investigate the protective role of YAP in TNF-αinduced intestinal epithelial barrier injury.Methods Caco-2 cells was cultured 14d to establish the intestinal mucosal barrier model,and then they were transfected with YAP plasmid and control plasmid vector,and 24h later they were exposed to TNF-α,and 48h later cell viability was quantified by CCK-8 assays.Cell cycle was determined by flow cytometric analysis.Cell migration was monitored by Transwell.Results Compared to the normal control group,after exposed to TNF-α,the growth rate of Caco-2 cells was significantly decreased,and the ratio of G0/G1 phase cells was significantly higher,while the S phase and G2/M phase proportion decreased,and markedly reduced cells proliferation.The over expression of YAP significantly ameliorated the decrease of the growth rate induced by TNF-α,and promoted the cell cycle in G0/G1 phase to S phase and G2/M phase transition.Transwell results showed that the number of cell migration in control group was(172.4±13.1),after adding TNF-αstimulation decreased significantly(80.4±9.3),while the number of cell migration induced by transfection of YAP was(124.2±9.2).The difference was statistically significant(P<0.01).Conclusion YAP inhibits intestinal epithelial barrier injury induced by TNF-αthrough regulating intestinal epithelial cell survival and migration.

关 键 词：肠黏膜屏障 Yes相关蛋白 肿瘤坏死因子-Α 细胞增殖 细胞迁移 

分 类 号：R574[医药卫生—消化系统]