干扰素α通过P204和RAS信号途径诱导大鼠血管平滑肌细胞凋亡  

Inducement Effect of Interferon Alpha on Apoptosis of Rat Vascular Smooth Muscle Cells via P204 and RAS Signal Pathway

作　　者：龙向淑[1] 吴强[1] 刘太河[2] 宋方[1] 黄晶[1] 田茂波[1] 肖燕[1] 

机构地区：[1]贵州省人民医院心血管内科,贵阳550002 [2]安顺学院数学与计算机科学学院,安顺561000

出　　处：《医药导报》2015年第12期1555-1558,共4页Herald of Medicine

基　　金：国家自然科学基金资助项目(81260030);贵州省科技攻关项目(黔科合LH字[2014]7023号)

摘　　要：目的观察干扰素-α(IFN-α)对大鼠血管平滑肌细胞(VSMCs)凋亡的影响,并探讨其部分机制。方法将VSMCs分为A、B、C 3组,A组转染非特异性siRNA,B组予IFN-α干预后转染非特异性siRNA,C组予IFN-α干预后转染IFN诱导蛋白204基因(IFI204)siRNA,均培养至48 h收集细胞。反转录聚合酶链反应(RT-PCR)法检测IFN诱导蛋白204(P204)mRNA表达,Western blot检测P204、RAS蛋白表达及RAF/ERK磷酸化变化,流式细胞仪Annexin-V FITC/PI法检测细胞凋亡。结果与A组比较,B组P204 mRNA和蛋白表达上调(P<0.05),细胞凋亡增多(P<0.05),伴RAS蛋白表达减少(P<0.05),RAF/ERK磷酸化水平下降(P<0.05);C组P204 mRNA及蛋白表达下调(P<0.05),细胞凋亡减少(P<0.05),RAS蛋白表达增多和RAF/ERK磷酸化水平上调(P<0.05)。结论 P204及RAS信号途径参与IFN-α诱导大鼠VSMCs凋亡过程的调控。Objective To investigate the effect of interferon alpha（ IFN-α） on apoptosis of vascular smooth muscle cells（ VSMCs） in rats and the related mechanism. Methods The cells were divided into three group： group A,group B and group C.Group A was transfected with nonspecific siRNA,group B was intervened with IFN-α and transfected with nonspecific siRNA,and group C was intervened with IFN-α and transfected with IFI204 siRNA. All the cells were cultured for 48 h. The expression of P204 mRNA was determined by semiquantitative reverse transcription polymerase chain reaction（ RT-PCR）.P204,RAS protein levels,and phosphorylation levels of RAF and ERK were analyzed by Western blotting. The cell apoptosis was analyzed by flow cytometry with Annexin-V FITC / PI method. Results As compared with group A,the expression of P204 mRNA and protein in group B was up-regulated（ P〈0.05）,the cell apoptosis was increased（ P〈0.05）,in the process of the above,the expression of RAS protein was decreased（ P〈0. 05） and the phosphorylation levels of RAF and ERK were dropped（ P〈0.05）.In group C,the expression levels of P204 mRNA and protein were down-regulated（ P〈0.05）,and cell apoptosis was decreased（ P〈0.05）,the expression of RAS protein and the phosphorylation levels of RAF and ERK were increased（ P〈0.05）.Conclusion P204 and RAS signal pathway participates in IFN-α regulation of apoptosis of VSMCs in rats.

关 键 词：干扰素 血管平滑肌细胞 干扰素诱导蛋白204 RAS信号途径 凋亡 

分 类 号：R965[医药卫生—药理学] R972[医药卫生—药学]