miR-24-3p靶向HIF-1α体外调节AngiotensinⅡ诱导的小鼠血管平滑肌细胞增殖、凋亡、迁移  

miR-24-3p-targeted HIF-1α in vitro regulation of proliferation,apoptosis and migration of rat vascular smooth muscle cells induced by AngiotensinⅡ

作　　者：张永安 徐建平 陈旭 龚绍慧 余斌 熊成英 詹剑 ZHANG Yong-An;XU Jian-Ping;CHEN Xu;GONG Shao-Hui;YU Bin;XIONG Cheng-Ying;ZHAN Jian(Department of Neurosurgery,Shangrao People′s Hospital,Shangrao 334000,China)

机构地区：[1]上饶市人民医院神经外科

出　　处：《中国免疫学杂志》2020年第1期31-35,共5页Chinese Journal of Immunology

基　　金：江西省卫生计生委科技计划项目(20167268)

摘　　要：目的:探讨miR-24-3p靶向HIF-1α对血管紧张素Ⅱ(AngiotensinⅡ)体外诱导的小鼠血管平滑肌细胞增殖、凋亡及迁移的影响。方法:分离C57BL/6小鼠原代动脉血管平滑肌细胞(VSMCs)体外培养,分为对照组和诱导组,诱导组用10-8 mol/L AngiotensinⅡ诱导VSMCs,采用qRT-PCR法检测miR-24-3p和HIF-1α表达,运用生物信息学预测和荧光素酶实验验证miR-24-3p和HIF-1α的靶向关系;体外原代培养VSMCs分为对照组,诱导组,诱导组+mimic,诱导组+pc-HIF-1α,采用Western blot法检测各组的HIF-1α蛋白表达水平,Colony forming法检测细胞增殖,Hoechest法检测细胞凋亡,划痕实验和Transwell小室迁移实验检测细胞迁移侵袭。结果:诱导组miR-24-3p mRNA的表达极显著低于对照组,HIF-1α的表达极显著高于对照组(P<0.01);HIF-1α是miR-24-3p的预测靶点;与对照组相比,诱导组和诱导组+pc-HIF-1α组HIF-1α的表达极显著增加,诱导组+mimic组HIF-1α的表达极显著下降(P<0.01);与对照组相比,诱导组和诱导组+pc-HIF-1α组细胞的集落率和迁移率显著上调、凋亡率显著下调,诱导组+mimic组细胞的集落率和迁移率显著下调、凋亡率显著上调。结论:miR-24-3p通过抑制HIF-1α的表达,抑制AngiotensinⅡ诱导的小鼠血管平滑肌细胞增殖和迁移,促进其凋亡。Objective:To explore the effects of miR-24-3p-targeted HIF-1αon proliferation,apoptosis and migration of rat vascular smooth muscle cells induced by AngiotensinⅡin vitro.Methods:Primary arterial vascular smooth muscle cells(VSMCs)of C57BL/6 rats were isolated and cultured in vitro,and they were divided into control group and induction group.Induction group was given 10-8 mol/L AngiotensinⅡto induce VSMCs,and the expression levels of miR-24-3p and HIF-1αwere measured by qRT-PCR,and bioinformatics prediction and luciferase assay were used to verify the targeting relationship between miR-24-3p and HIF-1α.In vitro primary cultured VSMCs were divided into control group,induction group,induction+mimic group and induction+pc-HIF-1αgroup,and Western blot was used to detect the expression level of HIF-1αprotein in each group,and Colony forming test was used to detect cell proliferation,and Hoechest method was used to detect apoptosis,and scratch test and Transwell chamber migration assay were used to detect cell migration and invasion.Results:The expression level of miR-24-3p mRNA in induction group was extremely significantly lower than that in control group,and the expression level of HIF-1αwas extremely significantly higher than that in control group(P<0.01).HIF-1αwas the predicted target of miR-24-3p.Compared with control group,the expression level of HIF-1αin induction group and induction+pc-HIF-1αgroup was extremely significantly increased while the expression of HIF-1αin induction group+mimic group was significantly decreased(P<0.01).Compared with control group,the colony rate and migration rate of cells were significantly up-regulated while the apoptosis rate was significantly down-regulated in induction group and induction+pc-HIF-1αgroup,and the colony rate and migration rate in induction+mimic group were significantly down-regulated while the apoptosis rate was significantly up-regulated.Conclusion:miR-24-3p inhibits the proliferation and migration and promotes the apoptosis of rat vascular smooth

关 键 词：HIF-1Α 动脉血管平滑肌细胞 细胞增殖 细胞凋亡 细胞迁移 

分 类 号：R363[医药卫生—病理学]