Hedgehog信号通路在缺氧肠上皮屏障功能调控的作用  

Regulatory effects of hedgehog pathway on intestinal epithelial barrier function under hypoxia

作　　者：杨光[1] 杨学超[1] 陈国庆[1] 孙力华[1] 杨桦[1] 

机构地区：[1]第三军医大学新桥医院普通外科,重庆400037

出　　处：《中华消化外科杂志》2013年第6期465-468,共4页Chinese Journal of Digestive Surgery

基　　金：国家自然科学基金重大国际合作项目（81020108023）;国家自然科学基金面上项目（30973113、81272078）;重庆市科委国际合作重点项目（201110008）,

摘　　要：目的探讨Hedgehog（Hh）信号通路在缺氧肠上皮屏障功能调控的作用。方法大鼠小肠上皮细胞系IEC-6细胞分为3组：常氧组（21％氧浓度）、缺氧组（2％氧浓度）、缺氧＋环巴胺组（用5mmol／L的环巴胺预处理30min后，再给予2％氧浓度进行缺氧处理）。RT-PCR检测Hh信号通路IHH、PTCH、GLI-1mRNA的表达变化，电阻测定仪检测跨上皮电阻（TER），Westernblot检测IHH、紧密连接蛋白经典表达分子胞质附着蛋白（ZO-1）、咬合蛋白（Occludin）、闭合蛋白（Claudin-1）的表达情况。组间比较采取单因素方差分析，两两比较采用LSD-t检验。结果RT-PCR检测结果表明：常氧组Hh信号通路IHH、PTCH、GLI-1mRNA的相对表达量分别为0．056±0．009、0．459±0．087、0．142±0．023；缺氧组分别为0．303±0．052、0．678±0．073、0．483±0．061，两组比较，差异有统计学意义（t=一14．05，一11．85，一6．52，P〈0．05）。Westernblot检测结果显示：常氧组和缺氧组的IHH相对蛋白表达量分别为0．39±0．06和0．91±0．15，两组比较，差异有统计学意义（t=一8．08，P〈0．05）。常氧组、缺氧组和缺氧＋环巴胺组的TER分别为（134±5）Ohm／cm。、（100±6）Ohm／cm2、（118±5）Ohm／cm2，3组比较，差异有统计学意义（F=1．0d，P〈0．05）。与常氧组比较，缺氧组下降约27．7％（t=7．84，P〈0．05）；与缺氧组比较，缺氧＋环巴胺组回升约16．4％，但仍低于常氧组（t=4．23，P〈0．05）。常氧组胞质附着蛋白一1、咬合蛋白、闭合蛋白一1的表达分别为1．184-0．24、0．80±0．13、0．90±0．09，缺氧组分别为0．58±0．08、0．32±0．05、0．50±0．09，缺氧＋环巴胺组分别为0．92±0．21、0．43±0．10、0．82±0．11，3组比较，差异有统计学意义（F=4．95，2．88，10．09，P〈0．05）。缺氧组较常氧组分别降低48．7％、40．0％、55．6％（t=12．86，Objective To investigate the regulatory effects of hedgehog pathway on intestinal epithelial barrier function under hypoxia. Methods The IEC-6 cells of rats were divided into 3 groups： the normoxia group （21% oxygen concentration）, the hypoxia group （2% oxygen concentration） and the hypoxia ＋ cyclopamine group （cells pretreated by 5 mmol/L of cyclopamine, and then exposed in an atmosphere with 2% oxygen concentration）. The mRNA expressions of IHH, PTCH and GLI-1 were detected, and the transepithelial electrical resistance （TER） was determined. The protein expressions of tight junction proteins （ ZO-1, Occludin, Clandin-1 ） and IHH were assayed by using the Western blot. All data were analyzed using the one-way analysis of variance or LSD-t test. Results The relative mRNA expressions of IHH, PTCH and GLI-1 were 0. 056 -＋ 0. 009, 0. 459±0. 087, 0. 142 ± 0.023 in the normoxia group, and 0.303 ± 0. 052, 0. 678 ± 0. 073, 0.483± 0.061 in the hypoxia group, with significant difference between the 2 groups （t = - 14. 05, - 11.85, -6. 52, P 〈0.05）. The relative protein expressions of IHH in the normoxia group and the hypoxia group were 0.39 ± 0.06 and 0.91 ±0.15, with a signi- ficant difference between the 2 groups （t = - 8. 08, P 〈 0.05 ）. The TERs of the normoxia group, the hypoxia group and the hypoxia ＋ cyclopamine group were （ 134± 5 ） Ohm/cm3, （ 100 ± 6 ） Ohm/cm3 and （ 118 ± 5 ） Ohm/cm3,with significant difference between the 3 groups （F = 1.04, P 〈 0.05 ）. Compared with the normoxia group, the TER of the hypoxia group was decreased by 27.7% （ t = 7.84, P 〈 0.05） ; compared with the hypoxia group, the TER of the hypoxia ＋ cyciopamine group were increased by 16.4% , but it was still significantly lower than the normoxia group （t = 4. 23, P 〈 0.05）. The expressions of ZO=I, Occludin and Claudin=l were 1.18±0.24, 0.80 ±0.13 and 0.90 ±0.09 in the normoxia group, and 0.58±0.08, 0.32±0.05 and 0.50 ±0.09 in the hypoxia group, and 0.92 �

关 键 词：HEDGEHOG信号通路 环巴胺 肠上皮屏障功能 跨上皮电子阻 紧密连接蛋白 

分 类 号：R[医药卫生]