过表达BIRC5基因对氧糖剥夺/复氧诱导的脑微血管内皮细胞活性及VEGF表达的影响  

Effect of over-expression of BIRC5 gene on the activity and VEGF expression of cerebral microvascular endothelial cells induced by oxygen glucose deprivation/reoxygenation

作　　者：黄建敏[1] 陈海燕[1] 云艳芳 杨桂新 蒋勇明 李晓岚 韦宝莹 周莹杰 彭立志 莫芬 李雪斌[1] Huang Jianmin;Chen haiyan;Yun Yanfang;Yang Guixin;Jiang Yongming;Li Xiaolan;Wei Baoying;Zhou Yingjie;Peng Lizhi;Mo Fen;Li Xuebin(Department of Neurology,The Affiliated Hospital of Youjiang Medical University For Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院附属医院神经内科,广西百色533000

出　　处：《右江民族医学院学报》2024年第1期1-6,12,共7页Journal of Youjiang Medical University for Nationalities

基　　金：国家自然科学基金项目(81860226)。

摘　　要：目的探讨过表达BIRC5基因对氧糖剥夺/复氧(OGD/R)诱导的小鼠脑微血管内皮细胞损伤的保护作用,并分析其可能的机制。方法将小鼠源性脑微血管内皮细胞,根据不同干预方式分为正常对照组(细胞正常培养)、细胞损伤组(细胞正常培养24 h,随后OGD3h/R3h损伤细胞)、BIRC5干预组(细胞预先转染腺病毒-BIRC5质粒并培养24 h,随后OGD3h/R3h处理)和阴性对照组(细胞预先转染腺病毒空质粒并培养24 h,随后OGD3h/R3h处理)。采用激光共聚焦显微镜观察各组细胞形态学变化;用MTT法和流式细胞仪分别检测各组细胞存活率和凋亡率;用RT-PCR和免疫印迹法分别检测各组细胞BIRC5和VEGF mRNA及蛋白表达。结果正常对照组的细胞骨架微丝彼此连接,分布规则,丝网状有序排列;细胞损伤组和阴性对照组的细胞微丝断裂,收缩变短或移向周边,微丝网状排列紊乱,可见细胞外形皱缩,间隙加大,少部分微丝缺失出现空隙;BIRC5干预组的细胞微丝连接,形态成长梭形,排列较规则,可见细胞间隙缩小,显示过表达BIRC5基因能够减轻损伤细胞的骨架微丝紊乱。与正常对照组相比,细胞损伤组、BIRC5干预组及阴性对照组细胞存活率降低,而细胞凋亡率增高,差异均有统计学意义(P<0.05);与细胞损伤组相比,BIRC5干预组的细胞存活率增高,而细胞凋亡率降低,差异有统计学意义(P<0.05)。与正常对照组相比,细胞损伤组、BIRC5干预组和阴性对照组的BIRC5和VEGF的mRNA及蛋白表达降低,差异有统计学意义(P<0.05);与细胞损伤组相比,BIRC5干预组细胞BIRC5和VEGF的mRNA及蛋白表达增高,差异有统计学意义(P<0.05)。结论过表达BIRC5基因对OGD/R诱导的脑微血管内皮细胞损伤具有保护作用,机制可能与上调VEGF表达有关,提示BIRC5调控VEGF表达促进血管新生可能是脑侧支循环建立和形成的重要机制之一。Objective To investigate the protective effect of over-expression of BIRC5 gene on microvascular endothelial cell injury induced by oxygen glucose deprivation/reoxygenation(OGD/R)in mouse brain,and to analyze its possible mechanism.Methods According to different intervention methods,mouse brain microvascular endothelial cells were divided into normal control group(normal cell culture),cell damage group(normal cell culture for 24 hours,followed by OGD3h/R3h injury),BIRC5 intervention group(The cells were transfected with BIRC5 plasmids of adenovirus and cultured for 24 hours and then underwent OGD for 3 hours/R for 3 hours.)and negative control group(The cells were transfected with empty plasmids of adenovirus and cultured for 24 hours and then underwent OGD for 3 hours/R for 3 hours.).Laser confocal microscope was used to observe the morphological changes of cells in each group.The cell survival rate and apoptosis rate were detected by MTT and flow cytometry,respectively.The mRNA and protein expressions of BIRC5 and VEGF were detected by RT-PCR and Western blot,respectively.Results The cytoskeletal microfilaments of the normal control group were connected with each other and regularly distributed in an orderly network.In the injury group and negative control group,the microfilaments of the cells were broken,and the contraction of the cells became shorter or moved to the periphery;the microfilaments were arranged in a disordered network;the cell appearance was shrunk,the gap enlarged and a small number of microfilaments missing with gaps;In the BIRC5 intervention group,the microfilaments of cells were connected,spindle shaped with regular arrangement and reduced gap between cells,indicating that the overexpression of BIRC5 gene could reduce the disorder of cytoskeletal microfilaments of damaged cells.Compared with the normal control group,the cell damage group,the BIRC5 intervention group and the negative control group had significantly reduced cell survival rates,but had significantly increased apoptosis rates,sh

关 键 词：过表达BIRC5基因 脑微血管内皮细胞 存活 凋亡 氧糖剥夺/复氧 血管内皮生长因子 

分 类 号：R743[医药卫生—神经病学与精神病学]