NO体外对肠上皮细胞表达紧密连接蛋白Occludin的影响
一氧化氮;Sin1;Caco-2细胞;紧密连接蛋白Occludin;免疫印迹;实时定量聚合酶链式反应,刘冬妍,崔巍,通讯作者:,EffectsofNOonthetightjunctionproteinoccludininintestin
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刘冬妍, 中国医科大学附属盛京医院检验科 辽宁省沈阳市 110004
崔巍, 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, 副教授, 博士, 主要从事感染免疫和肠道免疫.
通讯作者: 刘冬妍, 110004, 辽宁省沈阳市, 中国医科大学附属盛京医院检验科. dongyan.liu@yahoo.com.cn
电话: 024-81805756
收稿日期: 2007-04-17 修回日期: 2007-07-20
Effects of NO on the tight junction protein occludin in intestinal epithelial cells in vitro
Dong-Yan Liu, Wei Cui
Dong-Yan Liu, Department of Medical Laboratory, the Affiliated Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
Wei Cui, Department of Digestion Diseases, the Affiliated First Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
Correspondence to: Dong-Yan Liu, Department of Medical Laboratory, the Affiliated Shengjing Hospital of China Medical University, Shenyang 110001, Liaoning Province, China. dongyan.liu@yahoo.com.cn
Received: 2007-04-07 Revised: 2007-07-20
Abstract
AIM: To study the effects of NO on the intestinal mucosal barrier and on the tight junction protein occludin in intestinal epithelial cells in vitro.
METHODS: Colon cancer cell line (Caco-2 cells) was treated with Sin1, a NO donor, in a dose-dependent manner for 24 hours. The protein and total RNA of Caco-2 cells were extracted. Changes in occludin protein mRNA in Caco-2 cells stimulated by NO were determined by Western blotting and real-time quantitative pclymerase chain reaction, respectively.
RESULTS: The killing effect of NO on Caco-2 cells was dose-dependent. When treated with Sin1 at 125, 250, 500 and 1000 μmol/L doses, the levels of occludin protein (375 ± 0.5, 374 ± 0.8, 363 ± 0.3, 363 ± 0.7) and mRNA (0.689 ± 0.01, 0.578 ± 0.09, 0.554 ± 0.03, 0.619 ± 0.04) were significantly decreased compared with those in untreated Caco-2 cells (398 ± 0.7, 1, respectively, P < 0.01).
CONCLUSION: NO can directly kill intestinal epithelial cells ......
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