MEG3、miR-210在深低温停循环所致脑损伤中的作用  

Vitro experiments on MEG3-regulated miR-210 in the pathogenesis of deep hypothermic arrested circulation-induced brain injury

作　　者：刘顺军 温小华 罗阳 李铨华 LIU Shun-jun;WEN Xiao-hua;LUO Yang;LI Quan-hua(Ganzhou Municipal Hospital,Ganzhou,Jiangxi 341000)

机构地区：[1]赣州市立医院,江西赣州341000

出　　处：《赣南医科大学学报》2024年第10期993-998,1070,共7页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：赣州市指导性科技计划项目(GZ2023ZSF183)。

摘　　要：目的:通过体外实验探讨母系表达基因3(Maternally expressed gene 3,MEG3)、微小RNA-210(microRNA-210,miR-210)在深低温停循环(Deep hypothermic circulatory arrest,DHCA)所致脑损伤发病机制中的作用。方法:将PC12细胞分为对照组(Control组)、Si-MEG3-NC组、Si-MEG3_1组、Si-MEG3_2组和Si-MEG3_3组,严格按照干扰载体转染操作规程构建干扰表达MEG3的载体细胞,采用qRT-PCR检测干扰效率,选择干扰效率高的干扰表达MEG3的载体细胞进行实验。将PC12细胞分为对照组、氧-葡萄糖剥夺/复氧(Oxygen-glucose deprivation/reoxygenation,OGD/R)组,对OGD/R组细胞进行OGD/R模型构建,对Si-MEG3-NC细胞、Si-MEG3细胞进行OGD/R模型构建分别设为OGD/R+Si-MEG3-NC组、OGD/R+Si-MEG3组。⑴采用qRT-PCR检测各组细胞MEG3 mRNA及miR-210的表达;⑵采用CCK8法检测各组细胞增殖能力;⑶采用流式细胞术法检测各组细胞凋亡能力;⑷采用双荧光素酶实验检测各组细胞荧光素酶活性。结果:⑴与对照组比较,OGD/R组、OGD/R+Si-MEG3-NC组MEG3mRNA表达增加,OGD/R+Si-MEG3组MEG3 mRNA表达降低,差异有统计学意义(P<0.01);与OGD/R组比较,OGD/R+Si-MEG3组MEG3 mRNA表达降低,差异有统计学意义(P<0.01)。与对照组比较,OGD/R组、OGD/R+Si-MEG3-NC组miR-210表达降低,OGD/R+Si-MEG3组miR-210表达增加,差异有统计学意义(P<0.01);与OGD/R组比较,OGD/R+Si-MEG3组miR-210表达增加,差异有统计学意义(P<0.01)。⑵与对照组比较,OGD/R组、OGD/R+Si-MEG3-NC组细胞增殖能力降低,OGD/R+Si-MEG3组细胞增殖能力增加,差异有统计学意义(P<0.01);与OGD/R组比较,OGD/R+Si-MEG3组细胞增殖能力增加,差异有统计学意义(P<0.01)。⑶与对照组比较,OGD/R组、OGD/R+Si-MEG3-NC组细胞凋亡能力增加,OGD/R+Si-MEG3组细胞凋亡能力降低,差异有统计学意义(P<0.05);与OGD/R组比较,OGD/R+Si-MEG3组细胞凋亡能力降低,差异有统计学意义(P<0.01)。⑷Encori数据库预测显示,MEG3与miR-210存在互补结合位点,MEGObjective:To investigate the role of maternally expressed gene 3(MEG3)and miR-210 in the pathogenesis of brain injury caused by deep hypothermic circulatory arrest(DHCA)through in vitro experiments.Methods:PC12 cells were divided into the Control group,Si-MEG3-NC group,Si-MEG3_1 group,Si-MEG3_2 group,and Si-MEG3_3 group.We strictly followed the interference vector transfection operation procedures to construct the MEG3-interfering vector cells.qRT-PCR was used to detect the interference efficiency,and the vector cells with high MEG3 interference efficiency were selected for the experiment.PC12 cells were divided into the Control group and Oxygen-glucose deprivation/reoxygenation(OGD/R)group.The OGD/R group cells were subjected to OGD/R model construction,and the Si-MEG3-NC cells and Si-MEG3 cells were subjected to OGD/R model construction,which were designated as OGD/R+Si-MEG3-NC group and OGD/R+Si-MEG3 group,respectively.⑴qRT-PCR was used to detect the expression of MEG3 mRNA and miR-210 in each group of cells;⑵The CCK8 method was used to detect the proliferation ability of each group of cells;⑶Flow cytometry was used to detect the apoptosis ability of each group of cells;⑷The dual-luciferase assay was used to detect the luciferase activity of each group of cells.Results:⑴Compared with the Control group,the expression of MEG3 mRNA in the OGD/R group and OGD/R+Si-MEG3-NC group increased,while it decreased in the OGD/R+Si-MEG3 group,with statistically significant differences(P<0.01);compared with the OGD/R group,the MEG3 mRNA expression in the OGD/R+Si-MEG3 group decreased,with statistically significant differences(P<0.01).Compared with the Control group,the expression of miR-210 in the OGD/R group and OGD/R+Si-MEG3-NC group decreased,while it increased in the OGD/R+Si-MEG3 group,with statistically significant differences(P<0.01);compared with the OGD/R group,the miR-210 expression in the OGD/R+Si-MEG3 group increased,with statistically significant differences(P<0.01).⑵Compared with the Control group,t

关 键 词：微小RNA-210 母系表达基因3 深低温停循环 脑损伤 

分 类 号：R-33[医药卫生]