高糖抑制miR-126-5p促进肾小管上皮细胞损伤  

High glucose inhibition of miR-126-5p promotes renal tubular epithelial cell injury

作　　者：蒋琼[1,2] 杨婷 李兆飞[3] 邹艳 李青春 陈小辉[1,2] 何明杰 钟爱民[1,2] Jiang Qiong;Yang Ting;Li Zhaofei;Zhou Yan;Li Qingchun;Chen Xiaohui;He Mingjie;Zhong Aimin(Department of Nephrology,Jiangxi Provincial People′s Hospital,the First Affiliated Hospital of Nanchang Medical College,Nanchang 330006,China;Jiangxi Provincial Key Laboratory of Nephrology,Nanchang 330006,China;Department of Nephrology,Leping Tongkang Traditional Chinese Medicine Hospital,Leping 333399,China)

机构地区：[1]江西省人民医院肾内科(南昌医学院第一附属医院),南昌330006 [2]江西省肾脏病重点实验室,南昌330006 [3]乐平同康中医医院肾内科,乐平333399

出　　处：《中国医师杂志》2023年第12期1829-1834,共6页Journal of Chinese Physician

基　　金：江西省自然科学基金(20202BABL206027)。

摘　　要：目的探究糖尿病肾病(DN)疾病发展过程中的miRNA差异表达谱,并且进一步探讨miR-126-5p参与肾小管上皮细胞高糖诱导损伤的作用机制。方法首先,从基因表达综合数据库(GEO)中下载已有的芯片数据,依次使用GEO2R、miRanda、基因本体论(GO)分析以及京都基因与基因组百科全书(KEGG)分析对差异miRNA进行数据挖掘。随后,采用高糖诱导HK-2细胞损伤模型,再分为3组:高糖模型组、si-HOTAIR组、si-HOTAIR+miR-126-5p inhibitor组,对3组细胞依次转染siRNA-NC、siRNA-HOTAIR以及siRNA-HOTAIR+miR-126-5p mimic,并培养于含60 mmol/L葡萄糖的培养基中。流式细胞仪检测各组细胞凋亡水平变化,CCK-8检测细胞增殖变化。结果通过GEO进行数据挖掘分析发现,相较于普通小鼠,DN小鼠的肾脏组织中存在74个miRNA表达上调,80个miRNA表达下调,富集分析结果表明miRNA可以靶向Wnt、PKG、MAPK以及Rap1等信号通路,且miR-126-5p显著下调。高糖诱导HK-2细胞损伤模型中,实验结果表明,60 mmol/L高糖浓度下细胞增殖活性抑制效果较为显著(P<0.05);高糖刺激会显著降低miR-126-5p的表达(P<0.05)。流式细胞仪结果表明,与高糖模型组相比,si-HOTAIR组细胞凋亡率显著下降(P<0.05),而si-HOTAIR+miR-126-5p inhibitor组细胞凋亡率显著上升(P<0.05)。CCK-8实验表明,与高糖模型组相比,si-HOTAIR组的细胞活力显著上升(P<0.05);而si-HOTAIR+miR-126-5p inhibitor组细胞活力则被抑制(P<0.05)。结论miR-126-5p可以抑制高糖诱导HK-2细胞凋亡,保护HK-2细胞。Objective To explore the differential expression profile of miRN in the development of diabetes nephropathy(DN),and further explore the mechanism of miR-126-5p involved in high glucose induced injury of renal tubular epithelial cells.Methods Firstly,we downloaded existing chip data from the Gene Expression Integrated Database(GEO)and used GEO2R,miRanda,gene ontology(GO)analysis,and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis to mine differential miRNAs.Subsequently,a high glucose induced HK-2 cell injury model was used and divided into three groups:high glucose model group,si-HOTAIR group,and si HOTAIR+miR-126-5p inhibitor group.The three groups of cells were sequentially transfected with siRNA-NC,siRNA-HOTAIR,and siRNA-HOTAIR+miR-126-5p mimic,and cultured in a medium containing 60 mmol/L glucose.Flow cytometry was used to detect changes in apoptosis levels in each group,while cell counting kit-8(CCK-8)was used to detect changes in cell proliferation.Results Through data mining analysis using GEO,it was found that compared to ordinary mice,DN mice had 74 upregulated miRNAs and 80 downregulated miRNAs in their kidney tissue.Enrichment analysis results showed that miRNAs could target signaling pathways such as Wnt,PKG,MAPK,and Rap1,and miR-126-5p was significantly downregulated.In the high glucose induced HK-2 cell injury model,the experimental results showed that the inhibitory effect on cell proliferation activity was more significant at a high glucose concentration of 60 mmol/L(P<0.05);High glucose stimulation significantly reduced the expression of miR-126-5p(P<0.05).The results of flow cytometry showed that compared with the high glucose model group,the apoptosis rate of the si-HOTAIR group significantly decreased(P<0.05),while the apoptosis rate of the si-HOTAIR+miR-126-5p inhibitor group significantly increased(P<0.05).The CCK-8 experiment showed that compared with the high glucose model group,the cell viability of the si-HOTAIR group significantly increased(P<0.05);The cell viability of the si-HOT

关 键 词：肾小管上皮细胞 miR-126-5p 细胞凋亡 糖尿病肾病 

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