醌氧化还原酶2下调对大鼠主动脉平滑肌细胞线粒体膜电位的影响  

Effects of down-regulation of quinone reductase 2 on mitochondrial membrane potential in rat aortic smooth muscle cells

作　　者：蔡静波[1] 季鹏[1] 尹红丽[1] 王岚[1] 郭红梅[1] 

机构地区：[1]江苏省老年医学研究所心内科,江苏省南京市210024

出　　处：《实用老年医学》2015年第2期112-115,共4页Practical Geriatrics

基　　金：江苏省自然科学基金(BK2011841)

摘　　要：目的研究醌氧化还原酶2(NQO2)下调对大鼠主动脉平滑肌细胞(RASMCs)线粒体膜电位的影响。方法将RASMCs分为3组:野生型组、阴性对照组和NQO2下调组。利用RNA干扰技术下调体外培养的RASMCs中NQO2,采用5-溴脱氧尿嘧啶核苷(Brd U)渗入法检测细胞增殖,用阳离子染料JC-1标记RASMCs线粒体内膜并检测线粒体膜电位,试剂盒法检测总超氧化物歧化酶(SOD)活性和细胞色素C水平。结果在野生型组、阴性对照组和NQO2下调组中,Brd U阳性细胞比率分别为(18.94±1.43)%,(18.29±0.92)%和(9.21±1.42)%,NQO2下调组的细胞增殖明显被抑制(P<0.01);3组细胞经JC-1标记后红色/绿色荧光比值分别为15.48±0.41,15.96±0.52和13.58±0.12,NQO2下调组细胞线粒体膜电位明显下降(P<0.01)。细胞色素C水平在3组细胞中分别为(9.62±0.29)ng/L,(9.36±0.07)ng/L和(15.83±0.83)ng/L,NQO2下调组明显升高(P<0.01)。结论 NQO2的下调可降低RASMCs线粒体膜电位,致细胞色素C释放增加,抑制细胞增殖,诱导细胞凋亡,发挥抗动脉粥样硬化作用。Objective To investigate the effects of down-regulation of quinone reductase 2（ NQO2） on mitochondrial membrane potential in rat aortic smooth muscle cells（ RASMCs）. Methods RASMCs were divided into three groups,wild group,negative control group and NQO2down-regulated group. A lentiviral vector incorporating NQO2 short interference RNA of short hairpin was transduced into RASMCs. Cell proliferation was detected by bromodeoxyuridine（ Brd U） assay. Flow cytometry was used to observe mitochondrial membrane potential. The contents of superoxide dismuta（ SOD） and cytochrome C were detected by using kits. Results The proliferation of RASMCs was significantly inhibited by down-regulation of NQO2. Mitochondrial membrane potential in RASMCs with lower NQO2（ 13. 58 ± 0. 12） was obviously lower than that in wild（ 15. 48 ± 0. 41） and negative control cells（ 15. 96 ± 0. 52）（ P〈0. 01）. Compared with wild（ 9. 62 ± 0. 29 ng / L） and negative control cells（ 9. 36 ± 0. 07 ng / L）,RASMCs with suppressed NQO2 had significantly higher level of cytochrome C（ 15. 83 ± 0. 83 ng / L）（ P〈0. 01）. Conclusions This study suggests that down-regulation of NQO2 significantly suppresses RASMCs proliferation and mitochondrial membrane potential,which may be the mechanism against atherosclerosis.

关 键 词：线粒体膜电位 大鼠主动脉平滑肌细胞 醌氧化还原酶2 RNA干扰 凋亡 

分 类 号：R543.5[医药卫生—心血管疾病]