晚期氧化蛋白产物在小肠上皮细胞的上皮间质转化中的作用  

The role of advanced oxidation protein products in epithelial mesenchymal transition of small intestinal epithelial cells

作　　者：郑羽[1] 聂宗强 陈鸿源[1] 王襄瑜[1] 黄海啸[1] 黄良祥[1] 曾长青[1] Zheng Yu;Nie Zongqiang;Chen Hongyuan;Wang Xiangyu;Huang Haixiao;Huang Liangxiang;Zeng Changqing(Department of Gastrointestinal Surgery,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou 350001,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院胃肠外科,福州350001

出　　处：《中华炎性肠病杂志（中英文）》2020年第2期124-130,共7页Chinese Journal of Inflammatory Bowel Diseases

基　　金：福建省自然科学基金面上项目(2016J01504)。

摘　　要：目的探讨晚期氧化蛋白产物(AOPPs)在小肠上皮细胞的上皮间质转化(EMT)中的作用,为研究炎症性肠病(IBD)患者小肠纤维化的发病机制提供依据。方法体外培养小鼠小肠隐窝上皮细胞IEC-6,采用次氯酸氧化修饰大鼠血清白蛋白(RSA)的方法体外制备AOPPs。将IEC-6细胞分为AOPPs组、RSA组和空白对照组。采用50、100、200、400μg/ml的AOPPs干预IEC-6细胞作为AOPPs组,50、100、200、400μg/ml的RSA干预IEC-6细胞作为RSA组,空白对照组不做任何干预。干预24 h后观察细胞形态并采用流式细胞术检测细胞凋亡,统计分析凋亡率最高时AOPPs的浓度。采用该浓度的AOPPs干预IEC-6细胞,流式细胞术检测48、72 h的细胞凋亡并统计分析不同时间点凋亡率的差异。荧光定量PCR检测AOPPs组、RSA组和空白对照组E-cadherin、纤维连接蛋白(Fibronectin)及参与调节细胞EMT过程的转录因子Snail、Slug的mRNA表达以及细胞外基质胶原蛋白Ⅰ(CollagenⅠ)的mRNA表达。结果AOPPs可以诱导IEC-6细胞的凋亡,并具有时间及浓度依赖效应(P<0.05);400μg/ml的AOPPs干预72 h细胞的凋亡率最高,达到(17.30±1.03)%。AOPPs明显下调IEC-6细胞E-cadherin的mRNA表达,而明显上调Fibronectin及CollagenⅠ的mRNA表达,同时转录因子Snail和Slug mRNA表达也明显增加(均P<0.05)。结论AOPPs通过诱导小肠上皮细胞凋亡,上调细胞的转录因子Snail和Slug的基因表达,抑制E-cadherin的转录,同时上调Fibronectin和CollagenⅠ基因表达,促进肠上皮细胞EMT,在IBD小肠纤维化中发挥一定作用。Objective To investigate the role of advanced oxidation protein products(AOPPs)in epithelial-mesenchymal transition(EMT)of small intestinal epithelial cells,and provide basis for the study of mechanism of intestinal fibrosis(IF)in intestinal bowel disease(IBD).Methods The small intestinal crypt epithelial cells IEC-6 were cultured in vitro.AOPPs was made by hypochlorous acid oxidation of rat serum albumin(RSA)in vitro.IEC-6 cells were divided into blank control group,AOPPs group and RSA group.The different concentrations(50,100,200,400μg/ml)of AOPPs were used to interfere with IEC-6 cells respectively in AOPPs group,50,100,200,400μg/ml RSA were used to interfere with IEC-6 cells respectively in RSA group and no intervention in blank control group.The morphological changes of IEC-6 cells were observed and apoptosis was detected by flow cytometry in 3 groups 24 h after intervention.When the apoptosis rate was highest,the concentration of AOPPs were analyzed statistically and used to interfere IEC-6 for 48 and 72 h.The difference of apoptosis rate were analyzed between the different times.The mRNA expression of E-cadherin,Fibronectin,Snail,Slug and CollagenⅠwere detected in blank control group,AOPPs group and RSA group by fluorescence quantitative PCR.Results AOPPs could induce the apoptosis of IEC-6 cells with time-and concentration-dependent effects(P<0.05).The apoptosis rate was(17.30±1.03)%,which reached the peak by the intervention of 400μg/ml AOPPs for 72 h.AOPPs significantly decreased the mRNA expression of E-cadherin in IEC-6 cells,significantly increased the mRNA expression of Fibronectin and CollagenⅠ,and significantly increased the mRNA expression of transcription factor Snail and Slug(all P<0.05).Conclusions AOPPs can promote the ECT of small intestinal epithelial cells and play a role in IF of IBD by inducing the apoptosis of IEC-6 cells,inhibiting of the transcription of E-cadherin,up-regulating the gene expression of the transcription factors Snail and Slug,while increasing the gene expressio

关 键 词：晚期氧化蛋白产物 上皮间质转化 小肠上皮细胞 小肠纤维化 炎症性肠病 

分 类 号：R57[医药卫生—消化系统]