TNF-α对肠上皮细胞紧密连接蛋白表达的作用
肿瘤坏死因子-α;紧密连接蛋白claudin-1;Caco-2细胞;免疫荧光;免疫印迹;实时定量聚合酶链式反应,崔巍,刘沛,刘冬妍,马力,通讯作者:,Effectoftumornecrosisfactor-αonproteinexpressionof
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崔巍, 刘沛, 中国医科大学附属第一医院消化内科 辽宁省沈阳市 110001
刘冬妍, 马力, 中国医科大学盛京医院感染科实验室 辽宁省沈阳市 110001
崔巍, 在读博士生讲师, 中国医科大学附属第一医院消化内科, 主要从事肝脏病的临床和科研工作.
国家自然科学基金资助项目, No. 30670947
通讯作者: 刘沛, 110001, 辽宁省沈阳市南京街155号, 中国医科大学附属第一医院消化内科. syliupei2003@yahoo.com.cn
电话: 024-83956962 传真: 024-83956451
收稿日期: 2007-02-25 接受日期: 2007-03-31
Effect of tumor necrosis factor-α on protein expression of tight junction protein in intestinal epithelial cells
Wei Cui, Dong-Yan Liu, Li Ma, Pei Liu
Wei Cui, Pei Liu, Department of Digestive diseases, the First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
Dong-Yan Liu, Li Ma, Department of Infectious Diseases, the Second Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
Supported by National Natural Science Foundation of China, No. 97100252
Correspondence to: Pei Liu, Department of Digestive Diseases, the First Affiliated Hospital of China Medical University, 155 Nanjing North Street, Shenyang 110001, Liaoning Province, China. syliupei2003@yahoo.com.cn
Received: 2007-02-25 Accepted: 2007-03-31
Abstract
AIM: To explore the mechanism of intestinal epithelial barrier disruption induced by tumor necrosis factor alpha (TNF-α). We detected the expression of the tight junction protein claudin-1 in Caco-2 cells with TNF-α treatment.
METHODS: Caco-2 cells between passages 20 and 30 were cultured with DMEM including 20% fetal bovine serum and 2 mmol/L glutamine for 7 days. Cells were then treated with different concentrations of TNF-α (0, 10, and 100 μg/L) for 24 hours when they became fused. Protein was extracted from cells to prepare NP-40 detergent-soluble and insoluble protein fractions. Western blot was used to measure the protein levels of phosphorylated and unphosphorylated claudin-1. Indirect immunofluorescence was applied to the localization of claudin-1 ......
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