miR-218-5p靶向HMGB1参与脂多糖诱导的足细胞损伤  

miR-218-5p targeting HMGB1 is involved in LPS-induced podocyte injury

作　　者：于群 马启启 王倩惠 林建功[1] 李燕梅 郭齐[1] YU Qun;MA Qiqi;WANG Qianhui;LIN Jiangong;LI Yanmei;GUO Qi(Department of Nephrology,Shandong Provincial Hospital Affiliated to Shandong University,Ji’nan 250021,China;Department of Nephrology,Shandong Provincial Hospital Affiliated to Shandong First Medical University,Ji’nan 250021,China)

机构地区：[1]山东大学附属省立医院肾内科,济南250021 [2]山东第一医科大学附属省立医院肾内科,济南250021

出　　处：《免疫学杂志》2022年第2期108-115,共8页Immunological Journal

摘　　要：目的探究miR-218-5p、高迁移率蛋白B1(HMGB1)在脂多糖(LPS)诱导的小鼠足细胞损伤中的作用。方法将小鼠足细胞分为4组:正常对照组、LPS诱导组、HMGB1 shRNA+LPS组、miR-218-5p mimics+LPS组。Western blot和Real-time PCR分别检测各组HMGB1、TLR4、MyD88、TNF-α、IL-6、synaptopodin、ZO-1蛋白和mRNA表达水平;免疫荧光检测synaptopodin、ZO-1的表达和定位;Transwell检测足细胞迁移能力;双荧光素酶报告基因验证miR-218-5p和HMGB1的关系。结果LPS刺激后,HMGB1、TLR4、MyD88表达明显升高,炎性因子TNF-α、IL-6表达升高,而足细胞标记蛋白synaptopodin、ZO-1表达降低,同时足细胞迁移能力明显增强。沉默HMGB1或过表达miR-218-5p可降低TLR4、MyD88,TNF-α、IL-6表达,增加synaptopodin、ZO-1表达,抑制足细胞迁移能力,缓解足细胞损伤。荧光素酶报告基因检测提示miR-218-5p可直接作用于HMGB1的3’UTR区,调控HMGB1的翻译过程。结论HMGB1在LPS诱导的足细胞损伤中发挥重要作用,沉默HMGB1或过表达miR-218-5p可通过抑制HMGB1通路,减轻LPS诱导的足细胞炎症反应,缓解足细胞损伤。To investigate the role of miR-218-5p and HMGB1 in LPS-induced mouse podocyte injury,mouse podocytes were recruited and divided into control group,LPS-induced group,HMGB1 shRNA+LPS group,and miR-218-5p mimics+LPS group.The protein and mRNA expression levels of HMGB1,TLR4,MyD88,TNF-α,IL-6,synaptopodin and ZO-1 were detected by Western blotting and Real-time PCR;the expression and localization of synaptopodin and ZO-1 were detected by immunofluorescence;the migration ability of podocytes was detected by Transwell.The relationship between miR-218-5p and HMGB1 was verified by Dual luciferase reporter assay.Data showed that LPS stimulation could up-regulate the expression of HMGB1,TLR4,MyD88 and inflammatory factors TNF-α and IL-6,down-regulate the expression of synaptopodin and ZO-1,and enhance the migration ability of podocytes.Silence of HMGB1 or overexpression of miR-218-5p could reduce the expression of TLR4,MyD88,TNF-α and IL-6,increase the expression of synaptopodin and ZO-1,inhibit the migration of podocytes and alleviate podocyte injury.The Dual luciferase reporter assay suggested that miR-218-5p could directly target on the3’ UTR region of HMGB1 and regulate the translation of HMGB1.Therefore,HMGB1 plays an important role in LPS-induced podocyte injury.Taken together,silence of HMGB1 or overexpression of miR-218-5p can reduce LPS-induced podocyte inflammatory response and alleviate podocyte injury by inhibiting HMGB1 pathway.

关 键 词：脂多糖 足细胞 高迁移率蛋白B1 miR-218-5p 

分 类 号：R392.12[医药卫生—免疫学]