摘要
目的研究miR-129-5p对酒精诱导的肠上皮细胞屏障功能障碍的作用.方法实时定量PCR检测酒精性肝病患者和健康人乙状结肠活检组织以及酒精诱导的分化的Caco-2细胞中miR-129-5p的表达.然后将分化的Caco-2细胞分成4组:对照组、酒精处理组(50 mmol/L酒精处理1 h)、酒精联合miR-129-5p抑制物处理组(酒精处理前,先转染miR-129-5p抑制物)和酒精联合miR-129-5p抑制物对照组(酒精处理前,先转染miR-129-5p抑制物对照).酒精处理后使用电阻仪检测跨上皮电阻(TER)值,Western blot法检测紧密连接相关蛋白闭合蛋白(occludin)和闭锁小带蛋白1(ZO-1)表达.结果与健康人相比,酒精性肝病患者乙状结肠活检组织中miR-129-5p的表达升高.酒精诱导后分化的Caco-2细胞中miR-129-5p的表达也上调.与对照组相比,酒精处理组中TER值、occludin和ZO-1表达均降低;与酒精处理组相比,酒精联合miR-129-5p抑制物处理组中TER值、occludin和ZO-1表达均升高.结论miR-129-5p在酒精性肝病患者乙状结肠和酒精诱导的分化的Caco-2细胞中的表达均上调,下调miR-129-5p的表达可减轻酒精诱导的肠上皮细胞屏障功能障碍.
Objective To investigate the effect of miR-129-5p on alcohol-induced intestinal epithelial permeability.Methods Real-time quantitative PCR was used to detect the expression level of miR-129-5p in sigmoid colon biopsies and alcohol-induced differentiated Caco-2 cells from patients with alcoholic liver disease and healthy people.Then,the differentiated Caco-2 cells were divided into 4 groups:the control group,the alcohol group(treatment with 50 mmol/L alcohol for 60 minutes),alcohol combined with miR-129-5p inhibitor group(transfection with miR-129-5p inhibitor before alcohol treatment),and alcohol combined with miR-129-5p inhibitor control group(transfection with miR-129-5p inhibitor control before alcohol treatment).After the alcohol treatment,the transepithelial electrical resista nee(TER)was analyzed by a resista nee meter,and the protein levels relevant to tight junction(occludin and ZO-1)were determined by Western blotting.Results The expressiion level of miR-129-5p was elevated in the biopsies of the patients with alcoholic liver disease compared with that in the healthy controls.Alcohol treatment in creased miR-129-5p expressi on in the differe ntiated Caco-2 cells.Compared with the control group,TER and the protein levels of occludin and ZO-1 decreased in the alcohol group.Compared with the alcohol group,TER and the protein levels of occludin and ZO-1 were enhanced in the alcohol combined with miR-129-5p group;however,these changes were not altered in the alcohol combined with miR-129-5p inhibitor control group.Conclusion The miR-129-5p expression is elevated in sigmoid colon biopsies and alcohol-induced differentiated Caco-2 cells from patients with alcoholic liver disease.Inhibition of miR-129-5p can suppress alcohol-induced intestinal epithelial permeability.
作者
任莉
屈阿敏
刘亚萍
罗妙莎
HEN li;QV Amin;LIU Yaping;LUO Miaosha(Department of Gastroenterology,First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2019年第10期892-896,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
陕西省国际科技合作计划项目(2018KW-042)。
