RAS信号途径参与干扰素-α抑制大鼠血管平滑肌细胞增殖  

RAS signal pathway participates in interferon-α mediated inhibition of proliferation of vascular smooth muscle cells in rats

作　　者：龙向淑[1] 吴强[1] 宋方[1] 黄晶[1] 张林军[1] 

机构地区：[1]贵州省人民医院心血管内科,贵州贵阳550002

出　　处：《基础医学与临床》2014年第7期882-885,共4页Basic and Clinical Medicine

基　　金：国家自然科学基金项目(81260030);贵州省优秀科技教育人才省长专项资金[黔省专合字(2009)30号]

摘　　要：目的探讨RAS信号途径在干扰素-α(IFN-α)抑制大鼠血管平滑肌细胞(VSMCs)增殖中的作用。方法应用转染IFI204 siRNA和/或IFN-α瞬时干预体外培养的大鼠VSMCs,以非特异性siRNA转染组为对照组,用MTT法测定细胞活力,流式细胞仪分析细胞周期,RT-PCR法和Western blot分别检测P204 mRNA、P204和Ras蛋白的表达及RAF与ERK的磷酸化水平。结果与对照组比较,IFN-α组P204 mRNA和蛋白表达上调(P<0.01),细胞活力和细胞周期G1/S转换下调(P<0.01),伴RAS蛋白表达减少(P<0.01),RAF及ERK磷酸化水平下降(P<0.01);转染IFI204 siRNA后再加IFN-α干预,P204 mRNA及蛋白表达下调(P<0.01),而G0/G1期细胞增多,S期细胞减少(P<0.01),且RAS蛋白表达和RAF及ERK磷酸化水平亦下调(P<0.01)。结论 RAS信号途径参与IFN-α抑制大鼠VSMCs增殖。Objective To study the action of RAS signaling pathway in interferon alpha （IFN-α） mediated to inhibition proliferation of vascular smooth muscle cells （VSMCs） in rats.Methods Cultured VSMCs were treated with transfection IFI204 siRNA and/or IFN-α.Nonspecific siRNA transfection group was as control group.The cell vitality was detected by MTT method,and the cell cycle was analyzed by flow cytometry.The expression of P204 mRNA was determined by semiquantitative RT-PCR.P204,RAS protein and the phosphorylation levels of RAF and ERK was analyzed by Western blot.Results Compared with control group,the expression of P204 mRNA and protein up-regulated（P 〈 0.01）,the cell vitality and the cell cycle of G1/S transition down-regulated （P 〈 0.01）.In the process of the above,the expression of RAS protein decreased（P 〈0.01） and the phosphorylation levels of RAF and ERK droped（P 〈 0.01） ; When intervented with IFN-α after transfection IFI204 siRNA,down-regulated the expression of P204 mRNA and protein were down-regulated（P 〈 0.01）,and cells of G0/G1 stage increased and S stage decreased （P 〈0.01）,but the expression of RAS protein and the phosphorylation levels of RAF and ERK did not increase correspondingly（P 〈0.01）.Conclusions RAS signal pathway participates in interferon-α mediated inhibition of proliferation of vascular smooth muscle cells in rats.

关 键 词：干扰素 血管平滑肌细胞 增殖 干扰素诱导蛋白 P204 RAS信号途径 

分 类 号：R363[医药卫生—病理学]