miR-223在尿毒症肠组织的表达及对紧密连接蛋白表达的影响  

Expression of miR-223 in uremia intestinal tissue and its effect on expression of tight junction proteins

作　　者：何荃[1] 陈蕾[1] 魏萌[1] 魏丽敏 薛瑾虹 渠宁 刘华[1] HE Quan;CHEN Lei;WEI Meng;WEI Limin;XUE Jinhong;QU Ning;LIU Hua(Department of Blood Purification, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, China)

机构地区：[1]西安交通大学第一附属医院血液净化科,陕西西安710061

出　　处：《西部医学》2021年第2期180-185,共6页Medical Journal of West China

基　　金：陕西省重点研发计划-一般项目(2019SF-006);西安交通大学第一附属医院科研发展基金项目(2017QN-26)。

摘　　要：目的探讨miR-223在尿毒症肠上皮细胞的表达情况及对尿毒症肠上皮紧密连接蛋白表达的影响及机制。方法建立5/6肾切除尿毒症大鼠模型随机分为尿毒症组10只、尿毒症+益生菌组10只,同时设立假手术组10只作为对照组,通过RT-qPCR法检测肠组织miR-223和紧密连接蛋白(occludin、claudin-1和ZO-1)mRNA的表达;对体外培养的肠上皮Caco2细胞,采用脂质体介导的miR-223模拟物(miR-223-mimics)以及抑制剂(miR-223-inhibitor)和阴性对照转染体外培养的Caco2细胞,继而检测紧密连接蛋白的表达变化。结果与对照组相比较,miR-223、occludin、claudin-1、ZO-1 mRNA表达量在尿毒症组显著下降(P<0.05),而益生菌干预可以部分恢复上述基因表达;转染miR-223-mimics可以升高肠上皮Caco2细胞紧密连接蛋白occludin、claudin-1和ZO-1 mRNA和蛋白的表达水平,而转染miR-223-inhibitor得到相反的改变。结论尿毒症状态下应用益生菌可以上调肠上皮miR-223的表达及改善肠上皮紧密连接的破坏,提示miR-223表达下降可能是引起紧密连接蛋白表达下降的因素,尿毒症状态下改善miR-223的表达可能作为改善肠上皮细胞紧密连接破坏的新途径。Objective To investigate the expression of miR-223 in uremic intestinal epithelial cells(IECs)and its influence on the expression and mechanism of uremic intestinal epithelial tight junction proteins(TJPs).Methods The rats with uremia after 5/6 nephrectomy were randomly divided into uremia group(Group UR,n=10),uremia+probiotics group(Group UP,n=10),and sham operation group(Group SH,n=10)as control group.The expressions of miR-223,occludin,claudin-1 and ZO-1 were detected by RT-qPCR.In vitro cultured intestinal epithelial Caco2 cells were transfected with liposome-mediated miR-223-mimics,miR-223-inhibitor and negative control,and then the expression changes of TJPs were detected.Results Compared with the Group SH,the mRNA expression levels of miR-223,occludin,claudin-1 and ZO-1 were significantly decreased in the Group UR(P<0.05),while the probiotic intervention could partially restore the above gene expression.Transfection of miR-223-mimics increased the mRNA and protein expression of occludin,claudin-1 and ZO-1 in intestinal epithelial Caco2 cells,whereas transfection of miR-223-inhibitor resulted in reverse changes.Conclusion The application of probiotics in uremia can up-regulate the expression of miR-223 in the intestinal epithelium and improve the destruction of the tight junction of the intestinal epithelium,suggesting that the decreased expression of miR-223 may be the cause of the decreased expression of the TJPs,and the improved expression of miR-223 in uremia may be a new way to improve the destruction of the tight junction of the intestinal epithelial cells.

关 键 词：尿毒症 肠上皮细胞 miR-223 紧密连接蛋白 肠道屏障功能障碍 

分 类 号：R692.5[医药卫生—泌尿科学]