LncRNA-Gm4419对小鼠肾小球系膜细胞炎症、纤维化和增殖的影响  

lncRNA-Gm4419 on inflammation,fibrosis and proliferation induced by high glucose in mouse glomerular mesangial cells

作　　者：高清[1,2,3] 卢美琪 陈怡萱[3] 林梦舒 王科嘉 GAO Qing;LU Meiqi;CHEN Yixuan;LIN Mengshu;WANG Kejia(Department of Nephrology,Zhongshan Hospital of Xiamen University,Xiamen 361000,China;School of Medicine,Xiamen University,Xiamen 361000,China;The Third Clinical Medical College,Fujian Medical University,Fuzhou 350000,China)

机构地区：[1]厦门大学附属中山医院肾内科,厦门361000 [2]厦门大学医学院 [3]福建医科大学第三临床医学院

出　　处：《肾脏病与透析肾移植杂志》2023年第5期443-448,共6页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：福建省自然科学基金项目(2020J011206)。

摘　　要：目的:探讨在高糖条件下小鼠肾小球系膜细胞(MGMC)中lncRNA-Gm4419与miR-7214-3p结合对炎症、纤维化和增殖的影响。方法:qRT-PCR测定低糖组(5.5 mmol/L葡萄糖+19.5 mmol/L甘露醇)和高糖组(25 mmol/L葡萄糖)MGMC中miR-7214-3p的表达水平;生物信息学预测lncRNA-Gm4419与miR-7214-3p的结合位点;荧光素酶报告基因方法检测MGMC中lncRNA-Gm4419与miR-7214-3p的结合能力;qRT-PCR检测单核细胞趋化蛋白1(MCP-1)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、纤维蛋白(Fn)和胶原蛋白Ⅳ(Col.Ⅳ)的表达水平,流式细胞术和细胞计数试剂盒(CCK8)方法测定细胞增殖能力。结果:在高糖培养下,MGMC中miR-7214-3p的表达水平显著降低(P<0.05)。在低糖组中,沉默miR-7214-3p后MCP-1、TNF-α、IL-1β、Fn和Col.Ⅳ表达水平升高,系膜细胞增殖能力增强(P<0.05);在高糖组中,过表达miR-7214-3p后MCP-1、TNF-α、IL-1β、Fn和Col.Ⅳ表达水平降低,系膜细胞增殖能力减弱(P<0.05)。MGMC过表达lncRNA-Gm4419后MCP-1、TNF-α、IL-1β、Fn和Col.Ⅳ表达水平升高,系膜细胞增殖能力增强(P<0.05);同时过表达lncRNA-Gm4419和miR-7214-3p组较单独过表达lncRNA-Gm4419组,MCP-1、TNF-α、IL-1β、Fn和Col.Ⅳ表达水平降低,系膜细胞增殖能力减弱(P<0.05)。结论:在高糖条件下,lncRNA-Gm4419通过抑制miR-7214-3p的表达来促进系膜细胞的炎症反应、纤维化形成和增殖。Objective:To investigate the effects of lncRNA-Gm4419 binding with miR-7214-3p on inflammation,fibrosis,and proliferation in mouse glomerular mesangial cells(MGMCs)under high glucose conditions.Methodology:The expression of miR-7214-3p in MGMCs cultured in low glucose(5.5mmol/L glucose+19.5mmol/L mannitol)and high glucose(25mmol/L glucose)conditions was measured using qRT-PCR.The binding site of lncRNA-Gm4419 to miR-7214-3p was predicted by bioinformatics analysis.The binding ability of lncRNA-Gm4419 to miR-7214-3p in MGMCs was determined using the Luciferase reporter gene method.Additionally,the expression of monocyte chemotactic protein 1(MCP-1),tumor necrosis factor(TNF-α),interleukin-1 beta(IL-1β),fibronectin(Fn),and typeⅣcollagen(Col.Ⅳ)was assessed by qRT-PCR,and cell proliferation was evaluated using flow cytometry and Cell Counting Kit-8(CCK8)assays.Results:MiR-7214-3p expression significantly decreased in MGMCs cultured in high glucose.In the low glucose group,silencing miR-7214-3p led to increased expression of MCP-1,TNF-α,IL-1β,Fn,and Col.Ⅳ,as well as enhanced cell proliferation.Conversely,in the high glucose group,overexpression of miR-7214-3p resulted in decreased expression of MCP-1,TNF-α,IL-1β,Fn,and Col.Ⅳ,as well as decreased cell proliferation.Furthermore,overexpression of lncRNA-Gm4419 in MGMCs led to increased expression of MCP-1,TNF-α,IL-1β,Fn,and Col.Ⅳ,and enhanced cell proliferation.However compared to overexpression of lncRNA-Gm4419 alone,when both lncRNA-Gm4419 and miR-7214-3p were overexpressed,the expression of MCP-1,TNF-α,IL-1β,Fn,and Col.Ⅳdecreased,along with reduced cell proliferation.Conclusion:Under high glucose conditions,lncRNA-Gm4419 promote the inflammatory response,fibrosis,and proliferation of MGMCs by inhibiting the expression of miR-7214-3p.

关 键 词：长链非编码RNA lncRNA-Gm4419 miR-7214-3p 肾小球系膜细胞 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]