miR-4306通过MAPK/ERK信号通路抑制血管紧张素Ⅱ诱导的主动脉夹层的形成

miR-4306 Inhibits Aortic Dissection Induced by Angiotensin Ⅱ in Human Aortic Vascular Smooth Muscle Cells via MAPK/ERK Pathway

目的探讨miR-4306对血管紧张素Ⅱ(AngⅡ)诱导的主动脉血管平滑肌细胞增殖、迁移的影响及作用机制。方法将miR-4306mimic转染至人主动脉血管平滑肌细胞(HA-VSMC),按照细胞处理方式分为miRNA-NC组、miRNA-NC+AngⅡ组、miRNA-mimics组和miRNA-mimics+AngⅡ组;利用CCK-8细胞增殖法检测AngⅡ对HA-VSMC细胞的最佳作用时间、浓度以及miR-4306对AngⅡ诱导的HA-VSMC细胞增殖的影响,细胞划痕实验检测miR-4306对AngⅡ诱导的HA-VSMC细胞迁移的影响,Western blot检测HA-VSMC细胞MMP-9、MAPK/ERK信号通路相关蛋白表达。结果 AngⅡ呈剂量和浓度依赖性的方式诱导HA-VSMC细胞增殖,组间和不同时间点之间差异具有统计学意义(均P<0.05)。miR-4306过表达可以抑制AngⅡ诱导的HA-VSMC细胞增殖,与miRNA-NC+AngⅡ组比较,miRNA-mimics+AngⅡ组细胞增殖数量明显减少,组间差异具有统计学意义(P<0.05)。miR-4306过表达可以抑制AngⅡ诱导的HA-VSMC细胞迁移,与miRNA-NC+AngⅡ组比较,miRNA-mimics+AngⅡ组细胞迁移距离增加,组间差异具有统计学意义(P<0.01)。miR-4306过表达明显降低AngⅡ诱导的HA-VSMC细胞MMP-9蛋白和p-ERK1/2蛋白表达,与miRNA-NC+AngⅡ组比较,miRNA-mimics+AngⅡ组MMP-9、p-ERK1/2蛋白表达明显降低,组间差异具有统计学意义(均P<0.01)。结论 miR-4306可能通过MAPK/ERK信号通路抑制AngⅡ诱导的血管平滑肌细胞增殖、迁移功能。

Objective To explore the proliferation and migration effect of miR-4306 on AngⅡinduced human aortic vascular smooth muscle cell and its mechanism.Methods miR-4306 mimic was transfected to the HA-VSMC cells,and the cells were divided into miRNA-NC group,miRNA-NC＋AngⅡgroup,miRNA-mimics group and miRNA-mimics＋AngⅡ group according to the treatment methods.The optimum concentration and time of AngⅡtreating HA-VSMC cell were identified by CCK-8 assay,as well as effect of miR-4306 on AngⅡ induced proliferation of HA-VSMC cells.The migration effect of miR-4306 on AngⅡinduced migration of HA-VSMC cells was detected by cell wound scratch assay.Furthermore,the expression levels of MMP-9 and MAPK/ERK pathway relative protein were detected by Western blotting.Results AngⅡ significantly promoted HA-VSMC cell proliferation with time-and dose-dependent manner,and there were significant differences between the groups and different time points（P0.05）.Overexpression of miR-4306 inhibited HA-VSMC cell proliferation induced by AngⅡ,as compared to the miRNA-NC＋AngⅡ group,the proliferation cells were significantly decreased in the miRNA-mimics＋AngⅡgroup（P0.05）.Besides,overexpression of miR-4306 also inhibited HA-VSMC cell migration induced by AngⅡ,compared to the miRNA-NC＋AngⅡ group,the migration distance was increased significantly（P0.01）.Both MMP-9 and p-ERK1/2 protein was decreased remarkably after miR-4306 treatment with overexpression of miR-4306 in AngⅡ-mediated HA-VSMC cell,compared to the miRNA-NC＋AngⅡ group,the expression levels of MMP-9 and p-ERK1/2 protein were significantly decreased in the miRNA-mimics＋AngⅡ group（P0.01）.Conclusion miR-4306 inhibits proliferation and migration of human aortic vascular smooth muscle cells probably through MAPK/ERK signaling pathway.