miR-375靶向PI3K/AKT通路在高糖诱导的人视网膜内皮细胞增殖和血管生成中的作用  

Effect of micro ribonucleic acid-375 targeting phosphatidylinositol 3-kinase/protein kinase B pathway on high glucose-induced proliferation and angiogenesis of human retinal endothelial cells

作　　者：张悦之 殷小龙[1] 邓燕[1] 熊晓顺 ZHANG Yuezhi;YIN Xiaolong;DENG Yan;XIONG Xiaoshun(Ophthalmology Center of the Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi Province,China;Department of Laboratory Medicine,Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi Province,China)

机构地区：[1]南昌大学第二附属医院眼科中心,江西省南昌市330006 [2]南昌大学第二附属医院检验科,江西省南昌市330006

出　　处：《眼科新进展》2024年第2期89-93,共5页Recent Advances in Ophthalmology

基　　金：江西省自然科学基金项目(编号:20212BAB206049)。

摘　　要：目的探讨miR-375靶向磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路对高糖诱导的人视网膜内皮细胞(hRECs)增殖和血管生成的影响机制。方法体外培养hRECs,对hRECs进行转染及双荧光素酶检测。hRECs分为对照组、高糖组、高糖+miR-375组、高糖+miR-375+LM22B-10组。采用CCK-8实验检测细胞增殖能力,采用血管形成实验检测细胞血管形成能力,采用RT-qPCR检测hRECs内miR-375和PI3K mRNA的表达情况,采用Western blot检测hRECs内PI3K、p-AKT/AKT蛋白表达的情况。结果与对照组比较,高糖组、高糖+miR-375组、高糖+miR-375+LM22B-10组培养48 h和72 h时hRECs增殖活力、PI3K和p-AKT/AKT的蛋白表达水平、血管形成能力和PI3K mRNA表达均显著升高,而miR-375表达降低(均为P<0.05);与高糖组比较,高糖+miR-375组培养48 h和72 h时hRECs增殖活力、PI3K mRNA和蛋白以及p-AKT/AKT的蛋白表达水平、血管形成能力均降低,而miR-375表达升高(均为P<0.05);与高糖+miR-375组比较,高糖+miR-375+LM22B-10组培养48 h和72 h时hRECs增殖活力、血管形成能力和p-AKT/AKT蛋白表达水平均升高(均为P<0.05),而miR-375、PI3K mRNA差异均无统计学意义(均为P>0.05)。转染miR-375模拟物和PI3K野生型载体后,hRECs中相对荧光素酶活性分别较转染miR-375阴性对照、转染PI3K突变型载体后显著降低(均为P<0.05)。结论miR-375靶向抑制PI3K/AKT通路可抑制高糖诱导的hRECs增殖和血管生成,进而缓解DR。Objective To investigate the influencing mechanism of micro ribonucleic acid(miR)-375 targeting phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway on high glucose-induced proliferation and angiogenesis in human retinal endothelial cells(hRECs).Methods The hRECs were cultured in vitro,and transfection and dual luciferase assay were performed on them.These hRECs were divided into the control group,high glucose group,high glucose+miR-375 group,and high glucose+miR-375+LM22B-10 group.The Cell Counting Kit-8 was used to detect the cell proliferation ability,the angiogenesis assay was used to detect the vascular formation ability,real-time quantitative PCR was used to detect the miR-375 and PI3K mRNA expressions in hRECs,and Western blot was used to detect the PI3K and p-AKT/AKT protein expressions in hRECs.Results At 48 h and 72 h after the cultivation,compared with the control group,the proliferation viability,PI3K and p-AKT/AKT protein expressions,vascular formation ability,and PI3K mRNA expression in hRECs significantly increased,and the miR-375 expression in hRECs significantly decreased in the high glucose group,high glucose+miR-375 group and high glucose+miR-375+LM22B-10 group(all P<0.05).Compared with the high glucose group,the proliferation viability,PI3K mRNA and protein expressions,p-AKT/AKT protein expression and vascular formation ability in hRECs were significantly reduced,and miR-375 expression significantly increased in the high glucose+miR-375 group(all P<0.05).Compared with the high glucose+miR-375 group,the proliferation viability,vascular formation ability and p-AKT/AKT protein expression in hRECs significantly increased in the high glucose+miR-375+LM22B-10 group(all P<0.05);there was no significant difference in the miR-375 and PI3K mRNA(all P>0.05).After transfected with miR-375 mimic and wt-PI3K-pGL4,the relative luciferase activity in hRECs significantly decreased compared with transfection with miR-375 NC and mut-PI3K-pGL4(all P<0.05).Conclusion The targeted inhibition of the PI3K

关 键 词：高血糖 糖尿病视网膜病变 血管生成 miR-375 PI3K/AKT通路 

分 类 号：R774.1[医药卫生—眼科]