GRP78-ATF6-CHOP通路相关分子在葡聚糖硫酸钠诱导的小鼠结肠炎中的表达  

Expressions of GRP78-ATF6-CHOP pathway molecules in colonal tissue of dextran sulphate sodium-induced colitis mice

作　　者：郭腾飞 轩青霞 吴玉丹[2] 董仕桢 高磊[1] 陈攀[1] 常永超[2] 高强[1,3] GUO Tengfei;XUAN Qingxia;WU Yudan;DONG Shizhen;GAO Lei;CHEN Pan;CHANG Yongchao;GAO Qiang(College of Clinical Medicine,Henan University of Science and Technology,Department of Gastroenterology and Hepatology,the First Affiliated Hospital,Henan University of Science and Technology,Luoyang,Henan,471003;College of Clinical Medicine,Henan University of Science and Technology,Department of Clinical Laboratory,the First Affiliated Hospital,Henan University of Science and Technology,Luoyang,Henan,471003;Department of Gastroenterology and Hepatology,Beijing Rehabilitation Hospital of Capital Medical University,Beijing,100144)

机构地区：[1]河南科技大学临床医学院,河南科技大学第一附属医院消化内科,河南洛阳471003 [2]河南科技大学临床医学院,河南科技大学第一附属医院检验科,河南洛阳471003 [3]首都医科大学附属北京康复医院消化内科,北京100144

出　　处：《郑州大学学报（医学版）》2019年第6期806-811,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81370487)

摘　　要：目的:探讨GRP78-ATF6-CHOP通路在葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎中的表达及意义。方法:选取6-8周龄的健康清洁级129S1/SvJ种系小鼠,随机分为DSS组和对照组(n=8),DSS组饮用30 g/L DSS溶液,对照组饮用无菌蒸馏水;于第6天处死小鼠。通过疾病活动指数评分(DAI)、结肠长度测定和HE染色等方法评估小鼠肠道炎症程度,采用qRT-PCR检测小鼠结肠组织中炎症因子IL-1β、IL-6、TNF-α以及内质网应激相关分子葡萄糖调节蛋白78(GRP78)、IRE1α、PERK、活化转录因子6(ATF6)和CHOP mRNA的表达水平,采用免疫组化法和蛋白印迹法检测小鼠结肠组织中GRP78、ATF6和CHOP蛋白的表达。结果:对照组无肠炎表现,DSS组有严重的肠炎表现。与对照组相比,DSS组结肠组织中炎症因子IL-1β、IL-6、TNF-αGRP78、ATF6和CHOP mRNA表达升高(P均<0.001.。免疫组化显示GRP78主要表达于结肠上皮细胞的胞质及胞膜,ATF6主要表达于结肠上皮细胞的胞质及胞核,CHOP主要表达于结肠上皮细胞的胞核中;DSS组GRP78、ATF6和CHOP蛋白表达水平高于对照组(P均<0.001.。结论:GRP78-ATF6-CHOP通路相关分子可能在DSS诱导的小鼠结肠炎的发生、发展中起重要作用。Aim:To investigate the expressions of endoplasmic reticulum stress(ERS)-related molecules GRP78,ATF6 and CHOP in colonal tissue of dextran sulphate sodium(DSS)-induced colitis mice and analyze their significance in pathogenesis of colitis.Method:Six-to-eight-week-old 129S1/SvJ mice were randomly divided into DSS group and control group(n=8 for each group).Mice in DSS group were given 30 g/L DSS for 6 days.Mice in control group were given diatilled water.Disease Activity Index(DAI),colon length and inflammatory scores of colon were assessed.The mRNA levels of inflammatory cytokines IL-1β,IL-6 and TNF-α,and ERS-related molecules GRP78,IRE1α,PERK,ATF6 and CHOP were measured by qRT-PCR.The protein expressions of GRP78,ATF6 and CHOP in colonal tissue were evaluated by immunohistochemistry and Western blot.Results:There was no colitis in mice of control group,whereas severe colitis in DSS group.In DSS group,IL-1β,IL-6,TNF-α,GRP78,ATF6,CHOP mRNA and GRP78,ATF6,CHOP protein were all increased significantly compared with control group(all P<0.001).GRP78 protein was mainly expressed in cytoplasm and cytomembrane of colonic epithelial cells;ATF6 protein was mainly expressed in the nucleus and cytoplasm of colonic epithelial cells;CHOP protein was mainly expressed in the nucleus of colonic epithelial cells.Conclusion:The activity of GRP78-ATF6-CHOP pathway is involved in the pathogenesis of inflammatory bowel disease.

关 键 词：炎症性肠病 葡聚糖硫酸钠 葡萄糖调节蛋白78 活化转录因子6 CHOP 小鼠 

分 类 号：R574.6[医药卫生—消化系统]