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目的:研究线粒体融合素基因2(Mfn2)的突变体片段Mfn2-1A,对自发性高血压大鼠血管平滑肌细胞的增殖抑制作用,并进一步探讨相关作用机制。方法:用已构建的重组腺病毒Adv-Mfn2-1A感染大鼠血管平滑肌细胞,利用细胞计数、四甲基偶氮唑盐比色法检测血管平滑肌细胞增殖,流式细胞仪检测细胞生长周期,蛋白免疫印迹法分析Mfn2-1A对信号通路Mek-Erk1/2蛋白表达水平的影响。结果:重组腺病毒Adv-Mfn2-1A感染平滑肌细胞能正常表达相应蛋白;转染Adv-Mfn2-1A后能抑制平滑肌细胞增殖(P<0.05),第3天开始效果明显;Adv-Mfn2-1A较Adv-Mfn2作用效果更显著(P<0.05);转染Adv-Mfn2-1A和Adv-Mfn2后阻滞于G0/G1期的细胞明显增多,转染Mfn2-1A后阻滞在G0/G1期细胞达到(77.74±3.67)%;Mfn2-1A可降低平滑肌细胞中磷酸化Erk1/2蛋白的表达,效果比Mfn2更明显(P<0.05)。结论:Mfn2基因突变体片段1A可明显抑制血管平滑肌增殖,使细胞阻滞在G0/G1期,其机制与抑制Mek-Erk1/2信号通路有关。
AIM: To investigate the inhibitory effect of Mfn2-1A, a mutant of mitochondrial fusion gene 2 (Mfn2), on the proliferation of vascular smooth muscle cells in spontaneously hypertensive rats and to explore the underlying mechanisms. Methods: The rat vascular smooth muscle cells were infected with the recombinant adenovirus Adv-Mfn2-1A. Cell proliferation was measured by MTT assay and cell growth cycle by flow cytometry. The effect of Mfn2-1A on the protein expression of Mek-Erk1 / 2 was analyzed by Western blotting. Results: The recombinant adenovirus Adv-Mfn2-1A infected smooth muscle cells could express the corresponding protein normally. Adv-Mfn2-1A transfected cells could inhibit the proliferation of smooth muscle cells (P <0.05) and the effect was obvious on the third day. Adv-Mfn2-1A (P <0.05). Adv-Mfn2-1A and Adv-Mfn2 transfected cells showed a marked increase in cells arrested in G0 / G1 phase after transfection with Mfn2-1A, but they were blocked in G0 / G1 (77.74 ± 3.67)%. Mfn2-1A decreased the expression of phosphorylated Erk1 / 2 protein in smooth muscle cells, which was more significant than that of Mfn2 (P <0.05). CONCLUSION: 1A fragment of Mfn2 gene mutant can significantly inhibit the proliferation of vascular smooth muscle and block the cell cycle in G0 / G1 phase. Its mechanism is related to inhibition of Mek-Erk1 / 2 signaling pathway.