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目的研究睾酮对氧化型低密度脂蛋白(ox-LDL)诱导大鼠血管平滑肌细胞(VSMC)表型转化和增殖的抑制作用,并探讨其可能机制。方法培养大鼠VSMC,利用血清饥饿法进行同步化处理。将细胞分为对照组:不进行任何处理;ox-LDL组:给予50μg/m L ox-LDL处理;睾酮组:分别加入5×10-8mol/L和5×10-7mol/L睾酮预处理12 h,然后与50μg/m L ox-LDL共培养;另外设置血清组,用含100 m L/L胎牛血清的培养基培养。水溶性四甲基偶氮唑盐(WST-1)法检测各组细胞增殖的变化;流式细胞术检测各组细胞周期的变化;Western blot法检测各组细胞线粒体融合素2(Mfn2)、磷酸化的细胞外信号调节激酶1/2(p-ERK1/2)、增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)及骨桥蛋白(OPN)表达的变化。结果与对照组比较,ox-LDL组细胞增殖能力增强,G0/G1期细胞比例降低,S期细胞比例增加,Mfn2表达降低,p-ERK1/2表达增加,PCNA表达增加,α-SMA表达降低,OPN表达增加。与ox-LDL组比较,不同浓度睾酮组细胞增殖受抑制,G0/G1期细胞比例增加,S期细胞比例降低,Mfn2表达增加,p-ERK1/2表达降低,PCNA表达降低,α-SMA表达增加,OPN表达降低,以上作用呈现一定的浓度依赖性。结论睾酮可抑制ox-LDL诱导的大鼠VSMC发生表型转化及增殖,可能与其上调Mfn2抑制ERK1/2信号通路有关。
Objective To investigate the inhibitory effect of testosterone on the phenotypic changes and proliferation of rat vascular smooth muscle cells (VSMCs) induced by ox-LDL, and to explore its possible mechanism. Methods Rat VSMCs were cultured and synchronized by serum starvation. The cells were divided into control group: no treatment; ox-LDL group: 50μg / m L ox-LDL treatment; testosterone group: 5 × 10-8mol / L and 5 × 10-7mol / L testosterone pretreatment 12 h, then co-cultured with 50 μg / m L ox-LDL. Serum groups were also set up and cultured with 100 m L / L fetal bovine serum. The changes of cell proliferation in each group were detected by WST-1 method. The changes of cell cycle in each group were detected by flow cytometry. The expressions of Mfn2, The expression of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1 / 2), proliferating cell nuclear antigen (PCNA), α-smooth muscle actin and osteopontin (OPN) Results Compared with the control group, the proliferation of ox-LDL group was enhanced, the proportion of cells in G0 / G1 phase decreased, the proportion of S phase cells increased, the expression of Mfn2 decreased, the expression of p-ERK1 / 2 increased, , OPN expression increased. Compared with ox-LDL group, the proliferation of testosterone group was inhibited, the proportion of cells in G0 / G1 phase increased, the proportion of S phase cells decreased, the expression of Mfn2 increased, the expression of p-ERK1 / 2 decreased, Increase, OPN expression decreased, the above effect showed a certain concentration-dependent. Conclusion Testosterone can inhibit ox-LDL-induced VSMC phenotypic transformation and proliferation, which may be related to its up-regulation of ERK1 / 2 signaling pathway Mfn2.