miR-140-5p靶向HMGB1/IκB-α轴对糖尿病大鼠视网膜病变的影响  

Effect of miR-140-5p targeting HMGB1/IκB-αaxis on retinopathy of diabetic rats

作　　者：张梦瑶 牛姝 蔡静 赵志刚[1] 周艳[2] ZHANG Mengyao;NIU Shu;CAI Jing;ZHAO Zhigang;ZHOU Yan(the Second Department of Endocrinology,Shijiazhuang People's Hospital,Shijiazhuang 050000;Department of Clinical Laboratory,the Second Affiliated Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]石家庄市人民医院内分泌二科,河北石家庄050000 [2]河北医科大学附属第二医院检验科,河北石家庄050000

出　　处：《基础医学与临床》2023年第4期615-620,共6页Basic and Clinical Medicine

基　　金：石家庄市卫生健康委员会资助项目(20190161)。

摘　　要：目的探讨微小RNA(miRNA)miR-140-5p通过靶向调节高迁移率族蛋白1(HMGB1)/核因子κB抑制蛋白α(IκB-α)轴对糖尿病大鼠视网膜病变的影响。方法将糖尿病大鼠随机分为模型组、miR-140-5p激动剂(miR-140-5p agomir)组、激动剂阴性对照(agomir-NC)组,10只/组,另取10只正常大鼠作为对照组。分离血清及视网膜组织,RT-qPCR检测血清和视网膜组织miR-140-5p、HMGB1 mRNA表达水平;HE染色检测病理学变化;ELISA检测血清单核细胞趋化因子-1(MCP-1)、肿瘤坏死因子-ɑ(TNF-ɑ)、细胞间黏附分子-1(ICAM-1)含量;TUNEL染色检测视网膜细胞凋亡;双荧光素酶报告基因实验检测miR-140-5p、HMGB1的靶向关系;Western blot检测视网膜组织HMGB1、IκB-α、NF-κB p65蛋白表达。结果HMGB1为miR-140-5p的靶基因。模型组、agomir-NC组MCP-1、TNF-ɑ、ICAM-1含量、HMGB1、NF-κB p65表达、凋亡细胞及组织损伤较对照组增加,miR-140-5p agomir组较模型组降低(P<0.05);模型组、agomir-NC组miR-140-5p、IκB-α表达较对照组降低,miR-140-5p agomir组较模型组增加(P<0.05)。结论上调miR-140-5p可能通过靶向负调控HMGB1,提高IκB-α表达缓解糖尿病大鼠视网膜病变。Objective To investigate the effect of miR-140-5p on retinopathy in diabetic rats through targeted regulation of high mobility group protein 1(HMGB1)/nuclear factorκB inhibitory protein-α(IκB-α)axis.Methods Diabetic rats were randomly divided into model group,miR-140-5p agomir group,agomir-NC group,and control group with 10 in each.Serum and retinal tissue were isolated,and the expression of miR-140-5p and HMGB1 mRNA inserum and retinal tissue was detected by RT-qPCR.HE staining was used to evaluate pathological changes.The level ofserum monocyte chemoattractant protein-1(MCP-1),tumor necrosis factor-ɑ(TNF-ɑ)and intercellular adhesion molecule-1(ICAM-1)was detected by ELISA.Retinal cell apoptosis was examined by TUNEL staining.Dual luciferase reporter gene assay was used to detect the targeting relationship between miR-140-5p and HMGB1.The protein expressions of HMGB1,IκB-αand NF-κb p65 in retinal tissue were detected by Western blot.Results HMGB1 was the target gene of miR-140-5p.The content of MCP-1,TNF-ɑ,and ICAM-1,the expression of HMGB1 and NF-κB p65,apoptotic cells counting and tissue damage in the model group and agomir-NC group were found to be all higher than those in the control group.The miR-140-5p agomir group was lower than that in the model group(P<0.05).The expression of miR-140-5p and IκB-αin the model group and agomir-NC group were lower than those in the control group and the miR-140-5p agomir group was higher than that in the model group(P<0.05).Conclusions Up-regulation of miR-140-5p may negatively regulate HMGB1 and improve the expression of IκB-αto alleviate retinopathy of diabetic rats.

关 键 词：miR-140-5p 糖尿病视网膜病变 HMGB1/IκB-α 细胞凋亡 

分 类 号：R587.2[医药卫生—内分泌]