miR-142-3p在感染后肠易激综合征中的表达及意义  

Expression and significance of miR-142-3p in post-infectious irritable bowel syndrome

作　　者：林湫泠 张定国 熊锋 姚君 Lin Qiuling;Zhang Dingguo;Xiong Feng;Yao Jun(Department of General Practice,Shenzhen People’s Hospital(the First Affiliated Hospital,Southern University of Science and Technology,the Second Clinical Medical College,Jinan University),Shenzhen 518020,China;不详)

机构地区：[1]深圳市人民医院南方科技大学附属第一医院暨南大学第二临床医学院全科医学科,深圳518020 [2]深圳市人民医院南方科技大学附属第一医院暨南大学第二临床医学院消化内科,深圳518020

出　　处：《新医学》2022年第12期899-903,共5页Journal of New Medicine

基　　金：深圳市科创委项目(JCYJ20210324113803011)。

摘　　要：目的探讨微RNA(miR)-142-3p在感染后肠易激综合征(PI-IBS)中的表达及意义,进一步阐明miR-142-3p调控PI-IBS发生发展的分子机制。方法使用脂多糖(LPS)刺激后,在大鼠肠上皮细胞上考察miR-142-3p对高迁移率族蛋白B1(HMGB1)-Toll样受体4(TLR4)靶基因的作用;通过TargetScan预测软件分析发现HMGB1是miR-142-3p的靶基因,之后采用旋毛虫感染建立PI-IBS模型,原代肠上皮细胞进行小干扰RNA(siRNA)转染实验,提取肠上皮细胞总RNA,采用实时荧光定量PCR检测,检测细胞中miR-142-3p、HMGB1和TLR4的mRNA表达水平,考察miR-142-3p对HMGB1-TLR4靶基因的作用,并进一步验证HMGB1在miR-142-3p抗炎中的介导作用。结果炎症基因NOD样受体热蛋白结构域相关蛋白3(NLRP3)、IL-1β、IL-6、IL-18和TNF-α均为HMGB1-TLR4的靶基因。使用LPS刺激后,施加miR-142-3p大大抑制了HMGB1-TLR4靶基因(NLRP3、IL-1β、IL-6、IL-18和TNF-α)的表达(P均<0.01)。使用siRNA对肠上皮细胞中的HMGB1靶向敲低后,细胞中的HMGB1表达降低超过80%,HMGB1-TLR4靶基因(NLRP3、IL-1β、IL-6、IL-18和TNF-α)的表达均无明显变化(P均>0.05)。结论miR-142-3p在肠上皮细胞中具有抗炎作用,其抗炎作用依赖于HMGB1。Objective To explore the expression and significance of miR-142-3 p in post-infectious irritable bowel syndrome(PI-IBS),and to further clarify the molecular mechanism of miR-142-3 p in regulating the incidence and development of PI-IBSMethods After stimulation using the lipopolysaccharide(LPS),the effect of miR-142-3 p on high mobility group box 1(HMGB1)-toll-like receptor 4(TLR4)target genes was investigated in the rat intestinal epithelial cells.TargetScan prediction software analysis showed that HMGB1 was a target gene of miR-142-3 p.Subsequently,the PI-IBS model was established using trichinella infection.Primary intestinal epithelial cells were subjected to the siRNA transfection assay.Total RNA was extracted from intestinal epithelial cells.The expression levels of miR-142-3 p,HMGB1 and TLR4 mRNA in cells were quantitatively determined by real-time PCR.The effect of miR-142-3 p on the HMGB1-TLR4 target genes was evaluated.The mediating role of HMGB1 in the anti-inflammation of miR-142-3 p was further validated.Results Inflammatory genes NLRP3,IL-1β,IL-6,IL-18 and TNF-αwere all the target genes of HMCB1-TLR4.After the LPS stimulation,administration of miR-142-3 p significantly inhibited the expression levels pf HMGB1-TLR4 target genes(NLRP3,IL-1β,IL-6,IL-18 and TNF-α)(all P<0.01).After the targeted knockdown of HMCB1 in intestinal epithelial cells using siRNA,the HMGB1 expression level in the cells was down-regulated by more than 80%,whereas no significant changes were observed in the expression levels of HMGB1-TLR4 target genes(NLRP3,IL-1β,IL-6,IL-18 and TNF-α)(all P>0.05).Conclusion miR-142-3 p plays an anti-inflammatory role in intestinal epithelial cells,which is dependent on HMGB1.

关 键 词：感染后肠易激综合征 炎症 微RNA-142-3p 高迁移率族蛋白B1 

分 类 号：R574.4[医药卫生—消化系统]