【摘要】 目的探讨miR-425对人主动脉平滑肌细胞增殖迁移的作用及潜在的分子机制。方法实时定量RT-PCR(qRT-PCR)检测人主动脉平滑肌细胞(human aortic smooth muscle cells, HASMCs)中miR-425的表达水平。转染miR-342inhibitor改变HASMCs中内源性miR-425的表达,采用CCK-8法和流式细胞术检测细胞增殖及周期的变化,Transwell检测细胞迁移能力;Western blot检测PTEN、PI3K、p-AKT/AKT、MMP-9的蛋白表达水平。结果血管紧张素Ⅱ(angiotensionⅡ,AngⅡ)可时间及剂量依赖性地促进HASMCs中miR-425的表达。转染miR-425 inhibitor可抑制AngⅡ对HASMCs的促增殖作用,并抑制细胞迁移,同时细胞中PI3K、p-AKT/AKT及MMP-9的蛋白水平显著降低,PTEN水平显著升高。结论沉默miR-425可通过调控PTEN/PI3K/AKT信号通路抑制人主动脉平滑肌细胞的增殖及迁移,提示miR-425可作为血管重构的一个潜在诊疗靶点。更多还原

【Abstract】 Objective To investigate the regulatory effects of miR-425 on the proliferation and migration of human aortic smooth muscle cells(HASMCs) and the underlying molecular mechanisms. Methods miR-425 expression in human aortic smooth muscle cells was measured by real-time quantitative polymerase chain reaction(qRT-PCR). After down-regulation of miR-425 expression using miR-425 siRNA, HASMCs cell proliferation and cell cycle were detected by CCK-8 assay and flow cytometry, respectively. Transwell assay was used to measure cell migration ability. Furthermore, the protein expressions of PTEN, PI3 K, p-AKT/AKT and MMP-9 were measured by Western blot. Results Angiotensin II(Ang Ⅱ) promoted the expression of miR-425 in HASMCs in a time-and dose-dependent manner. miR-425 knockdown was able to reverse the Ang Ⅱ-induced cell proliferation and inhibit cell migration. Furthermore, the expressions of PI3 K, p-AKT/AKT and MMP-9 at protein level significantly decreased while PTEN expression increased after knocking down miR-425. Conclusion Silencing miR-425 inhibited HASMCs proliferation and migration likely by regulating PTEN/PI3 K/AKT signal pathway, suggesting that miR-425 may be a potential therapeutic target of vascular remodeling.更多还原