微RNA-140-5p靶向调节血管内皮生长因子表达对高糖诱导的人视网膜血管内皮细胞增殖、迁移和管腔形成的影响  

Effect of microRNA-140-5p targeted regulating vascular endothelial growth factor expression on proliferation,migration and lumen formation of human retinal vascular endothelial cells induced by high glucose

作　　者：郑洪祥 李庆航 李亚男 蒋曦曦 吕建平[1] 张婷娉 王淼[1,2] ZHENG Hongxiang;LI Qinghang;LI Yanan;JIANG Xixi;LYU Jianping;ZHANG Tingping;WANG Miao(Department of Ophthalmology,Jining NO.2 People′s Hospital,Jining 272000,Shandong Province,China;Department of Ophthalmology,Yingjisha County People′s Hospital,Yingjisha 834800,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]济宁市第二人民医院眼科,山东济宁272000 [2]英吉沙县人民医院眼科,新疆英吉沙834800

出　　处：《新乡医学院学报》2023年第10期917-925,共9页Journal of Xinxiang Medical University

基　　金：新疆维吾尔自治区自然科学基金资金资助项目(编号:2020D01A127)。

摘　　要：目的 探讨微RNA(miR)-140-5p靶向调节血管内皮生长因子(VEGF)表达对高糖诱导的人视网膜血管内皮细胞(hRECs)增殖、迁移和管腔形成的影响。方法 将对数生长期hRECs细胞分为正常对照组、高糖组、miR-140-5p组、阴性对照(NC)组、高糖+miR-140-5p组。正常对照组和高糖组细胞不做任何转染,miR-140-5p组和高糖+miR-140-5p组细胞转染miR-140-5p模拟物(miR-140-5p mimics),NC组细胞转染阴性对照模拟物(mimics NC)。高糖组和高糖+miR-140-5p组细胞用含30 mmol·L^(-1)葡萄糖的培养基制备高糖模型。应用实时荧光定量聚合酶链式反应法检测各组细胞中miR-140-5p表达水平,噻唑蓝法检测各组细胞增殖能力,Transwell法检测各组细胞迁移能力,管腔形成实验检测各组细胞成管能力,Western blot法检测各组细胞中VEGF蛋白的表达。将40只Sprague Dawley大鼠随机分成正常对照组、糖尿病模型组、糖尿病+NC组、糖尿病+miR-140-5p组,每组10只。除正常对照组外,其余各组大鼠腹腔注射65 mg·kg^(-1)链脲佐菌素制备糖尿病模型,正常对照组和糖尿病模型组大鼠尾静脉注射200μL生理盐水,糖尿病+miR-140-5p组大鼠尾静脉注射200μL miR-140-5p,糖尿病+NC组大鼠尾静脉注射200μL mimics NC,每日1次,持续给药7 d;再继续饲养8周,大鼠麻醉后,取视网膜,苏木精-伊红染色观察miR-140-5p对糖尿病模型大鼠视网膜血管生成的影响。结果 高糖组、高糖+miR-140-5p组细胞中miR-140-5p相对表达量显著低于正常对照组,miR-140-5p组细胞中miR-140-5p相对表达量显著高于正常对照组(P<0.01);NC组与正常对照组细胞中miR-140-5p相对表达量比较差异无统计学意义(P>0.05)。miR-140-5p组、NC组、高糖+miR-140-5p组细胞中miR-140-5p相对表达量显著高于高糖组(P<0.01)。NC组、高糖+miR-140-5p组细胞中miR-140-5p相对表达量显著低于miR-140-5p组(P<0.01)。高糖+miR-140-5p组细胞中miR-140-5p相对表达量显著低于NC组(Objective To investigate the effect of microRNA(miR)-140-5p targeted regulating vascular endothelial growth factor expression on proliferation,migration and lumen formation of human retinal vascular endothelial cells(hRECs)induced by the high glucose.Methods The hRECs at logarithmic growth phase were divided into the normal control group,high glucose group,miR-140-5p group,negative control(NC)group and high glucose+miR-140-5p group.The cells in the normal control group and high glucose group did not recieve any transfection.The cells in the miR-140-5p group and high glucose+miR-140-5p group were transfected with miR-140-5p mimics.The cells in the NC group were transfected with negative control mimics(mimics NC).The cells in high glucose group and high glucose+miR-140-5p group were used to prepare high glucose models by using a culture medium containing 30 mmol·L^(-1) glucose.The expression level of miR-140-5p in cells in each group was detected by real time fluorescence quantitative polymerase chain reaction.The proliferation ability of cells in each group was detected by thiazole blue method.The migration ability of cells in each group was detected by Transwell method.The tubular ability of cells in each group was detected by lumen formation experiment.The expression of VEGF in the cells in each group was detected by Western blot method.Forty Sprague Dawley rats were randomly divided into the normal control group,diabetes model group,diabetes+NC group,diabetes+miR-140-5p group,with 10 rats in each group.Except for the normal control group,rats in the other groups were intraperitoneally injected with 65 mg·kg^(-1) streptozotocin to prepare the diabetes model,and rats in the normal control group and the diabetes model group were injected with 200μL normal saline through the tail vein,the rats in the diabetes+miR-140-5p group were injected with 200μL miR-140-5p through the tail vein,the rats in the diabetes+NC group were injected with 200μL mimics NC,once a day for 7 days,and then continued to be fed for 8 wee

关 键 词：微RNA-140-5p 血管内皮生长因子 视网膜血管病变 人视网膜血管内皮细胞 

分 类 号：R587.2[医药卫生—内分泌]