siRNA沉默ARC对低氧下大鼠肺动脉平滑肌细胞活力的影响  

Effects of ARC siRNA on growth of rat pulmonary artery smooth muscle cells under hypoxic condition

作　　者：张婕 金泉 吴素玲 管明 ZHANG Jie;JIN Quan;WU Suling;GUAN Ming(The Second Department of Internal Medicine,Hangzhou Children′s Hospital,Hangzhou,Zhejiang 310014,China;Department of Otorhinolaryngology,Hangzhou First People′s Hospital,School of Medicine,Zhejiang University,Hangzhou,Zhejiang 310006,China)

机构地区：[1]浙江省杭州市儿童医院内二科,310014 [2]浙江大学医学院附属杭州市第一人民医院耳鼻喉科,310006

出　　处：《重庆医学》2019年第16期2723-2727,2733,共6页Chongqing medicine

基　　金：浙江省医药卫生科技计划项目(2015KYB294)

摘　　要：目的利用小干扰RNA(siRNA)技术干扰大鼠肺动脉平滑肌细胞(PASMCs)含胱冬肽酶富集功能域的凋亡抑制因子(ARC)的表达,验证ARC在PASMCs活力变化中的作用。方法体外培养PASMCs,分为常氧组、缺氧组、siRNA-Negative+缺氧组、siRNA-ARC+缺氧组。采用CCK-8法检测细胞活力情况,免疫荧光染色法及Western blot检测ARC在PASMCs中的表达,RT-PCR法分别检测caspase-3、Fadd、Bcl-2、Bad mRNA表达。结果低氧下PASMCs活力较常氧下增加,并呈时间依赖性;与siRNA-Negative+缺氧组比较,siRNA-ARC+缺氧组的PASMCs活力受到抑制;siRNA-ARC+缺氧组PASMCs的caspase-3、Bad、Fadd mRNA表达较siRNA-Negative+缺氧组增加,而Bcl-2 mRNA表达较siRNA-Negative+缺氧组减少(P<0.05)。结论ARC参与调控低氧下大鼠PASMCs生长,对PASMCs活力具有一定的增强作用,可能是促凋亡因子及抗凋亡因子共同作用的结果。Objective To observe whether ARC is involved in the growth of pulmonary arterial smooth muscle cells(PASMCs)under hypoxic condition through interfering the expression of apoptosis repressor with caspase recruitment domain(ARC)by siRNA.Methods PASMCs were cultured in vitro and divided into four groups:the normoxic group(under normoxia condition),the hypoxia group(under hypoxia condition),the siRNA-Negative+hypoxia group(transfected with siRNA-Negative under hypoxia condition)and the siRNA-ARC+hypoxia group(transfected with siRNA-ARC under hypoxia condition).The viability of the PASMCs was detected by CCK-8 assay.The expression level of ARC in PASMCs was measured by immunofluorescent staining and Western blot.The expression levels of caspase-3,Fadd,Bcl-2 mRNA and Bad mRNA were detected by RT-PCR.Results Compared with undering normoxia condition,the viability of the PASMCs undering hypoxic condition was increased in a time-dependent manner.Compared with the siRNA-Negative+hypoxia group,the cell viability and expression level of Bcl-2 mRNA in the siRNA-ARC+hypoxia group was decreased,while the expression levels of caspase-3,Bad and Fadd mRNA in the siRNA-ARC+hypoxia group were increased(P<0.05).Conclusion ARC is involved in the regulation of the growth of PASMCs in rats under hypoxia,and may have a certain enhancement effect on the activity of PASMCs,which may be the result of the combination effect of pro-apoptotic factors and anti-apoptotic factors.

关 键 词：低氧 含胱冬肽酶富集功能域的凋亡抑制因子 大鼠肺动脉平滑肌细胞 细胞活力 

分 类 号：R392.1[医药卫生—免疫学]