apelin抑制肺动脉平滑肌细胞增殖的分子机制研究

Molecular Mechanism of apelin Inhibiting Proliferation of Pulmonary Arterial Smooth Muscle Cells

目的:明确apelin对PASMC增殖的影响,并探讨相关的分子机制,为肺动脉高压新药的研发提供一定的理论和实验依据。方法:体外培养大鼠PASMC,采用TGF-β1刺激原代PASMC增殖,给予apelin干预,应用Western blot检测细胞中Smad2/3的磷酸化水平以及cyclin D1的蛋白水平,BrdU掺入法检测各组细胞增殖情况。结果:apelin干预组Smad2/3的磷酸化水平低于TGF-β1刺激组(P<0.05)。APJ siRNA转染组Smad2/3的磷酸化水平高于apelin干预组(P<0.05)。apelin干预组cyclin D1蛋白水平低于TGF-β1刺激组(P<0.05)。SB431542干预组cyclin D1蛋白水平高于apelin干预组(P<0.05)。apelin干预组PASMC增殖率低于TGF-β1刺激组(P<0.05)。APJ siRNA转染组、SB431542干预组及cyclin D1 siRNA转染组PASMC增殖率均高于apelin干预组(P<0.05)。结论:apelin可通过与其受体APJ结合,进而调节TGF-β1/Smad/cyclin D1轴抑制PASMC的增殖。apelin对PASMC增殖的抑制作用以及潜在机制为研发新的靶向药物提供了思路。

Objective:To clarify the effect of apelin on the proliferation of PASMC,and to explore the related molecular mechanism,and to provide a theoretical and experimental basis for the research and development of new drugs for pulmonary hypertension.Method:PASMC was cultured in vitro,and the proliferation of primary PASMC was stimulated by TGF-β1,and apelin intervention was given.The phosphorylation level of Smad2/3 and the protein level of cyclin D1 in the cells were detected by Western blot.BrdU incorporation method was used to detect cell proliferation in each group.Result:The phosphorylation level of Smad2/3 in apelin intervention group was lower than that in TGF-β1 stimulation group(P<0.05).Phosphorylation level of Smad2/3 in APJ siRNA transfection group was higher than that in apelin intervention group(P<0.05).The level of cyclin D1 protein in apelin intervention group was lower than that in TGF-β1 stimulation group(P<0.05).cyclin D1 protein level in SB431542 intervention group was higher than that in apelin intervention group(P<0.05).Proliferation rate of PASMC in apelin intervention group was lower than that in TGF-β1 stimulation group(P<0.05).The proliferation rate of PASMC in APJ siRNA transfection group,SB431542 intervention group and cyclin D1 siRNA transfection group were higher than that in apelin intervention group(P<0.05).Conclusion:apelin can regulate the TGF-β1/Smad/cyclin D1 axis to inhibit the proliferation of PASMC by binding to its receptor APJ.The inhibitory effect of apelin on the proliferation of PASMC and its potential mechanism provide an idea for the development of new targeted drugs.