下调动脉粥样硬化斑块中TRPC5的表达对人巨噬细胞凋亡的影响及机制研究  

TRPC5 expression in atherosclerotic plaque and effect of inhibition of its expression on human macrophage apoptosis

作　　者：闫博阳 赵津璋 陈虹[1] YAN Bo-Yang;ZHAO Jin-Zhang;CHEN Hong(Jiamusi City Center Hospital,Heilongjiang Province,Jiamusi 154002,China)

机构地区：[1]黑龙江省佳木斯市中心医院,佳木斯154002

出　　处：《中国免疫学杂志》2018年第10期1472-1477,共6页Chinese Journal of Immunology

摘　　要：目的:探讨动脉粥样硬化病变时瞬时受体电位通道5(TRPC5)基因表达对人巨噬细胞凋亡的影响。方法:建立小鼠动脉粥样硬化模型,通过RT-PCR检测动脉粥样硬化斑块TRPC5基因表达;将巨噬细胞分为对照组、NC组、ox-LDL组、TRPC5-siRNA组和TRPC5-siRNA+ox-LDL组,细胞处理24 h后Western blot检测各组细胞中TRPC5的蛋白表达; CCK8法及流式细胞术分别检测细胞的增殖及凋亡; Western blot检测凋亡蛋白含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase3)及丝氨酸苏氨酸激酶(AKT)信号通路AKT及磷酸化的AKT的蛋白表达。结果:成功建立动脉粥样硬化模型。TRPC5基因在动脉粥样硬化组中的表达显著高于对照组(P<0. 05);与对照组比较,TRPC5-siRNA组TRPC5、Caspase3的表达及细胞凋亡率显著降低,细胞增殖及p-AKT表达显著升高,ox-LDL组TRPC5、Caspase3的表达及细胞凋亡率显著升高,细胞增殖及p-AKT表达显著降低(P<0. 05); TRPC5-siRNA+ox-LDL组TRPC5、Caspase3的表达及细胞凋亡率显著低于ox-LDL组,高于TRPC5-siRNA组,细胞增殖及p-AKT表达显著高于ox-LDL组,低于TRPC5-siRNA组(P<0. 05); AKT在各组间表达差异无统计学意义(P>0. 05)。结论:TRPC5基因在动脉粥样硬化斑块中表达升高,下调TRPC5表达可减弱ox-LDL对巨噬细胞增殖的抑制作用和凋亡的促进作用,机制与AKT信号通路有关。Objective： To investigate the effect of TRPC5 gene expression in atherosclerosis on apoptosis of human macrophages. Methods： The mice atherosclerosis model was established,TRPC5 gene expression was detected in atherosclerotic plaques by RT-PCR; macrophages were divided into control group,NC group,ox-LDL group,TRPC5-siRNA group and TRPC5-siRNA＋ox-LDL group,the expression of TRPC5 protein was detected by Western blot cells were treated for 24 h; CCK8 assay and flow cytometry were used to detect cell proliferation and apoptosis; the expression of Caspase3,AKT,p-AKT protein were detected by Western blot. Results： The atherosclerotic model was established. The expression of TRPC5 gene in atherosclerosis group was significantly higher than that of control group（P〈0. 05）; compared with the control group,the expression of TRPC5,Caspase3 and cell apoptosis rate in TRPC5-siRNA group significantly decreased,cell proliferation and p-AKT expression increased significantly,the expression of TRPC5,Caspase3 and cell apoptosis rate in ox-LDL group significantly increased,cell proliferation and p-AKT expression significantly decreased（P〈0. 05）; the expression of TRPC5,Caspase3 and cell apoptosis rate in TRPC5-siRNA＋ox-LDL group was significantly lower than oxLDL group,higher than that of TRPC5-siRNA group,cell proliferation and p-AKT expression was significantly higher than that of oxLDL group,was lower than that of TRPC5-siRNA group（P〈0. 05）; AKT expression in each group showed no significant difference（P〈0. 05）. Conclusion： The expression of TRPC5 gene is increased in atherosclerotic plaques,and down regulation of TRPC5 expression can reduce the inhibitory effect of ox-LDL on the proliferation of macrophages and promote the apoptosis. The mechanism is related to the AKT signaling pathway.

关 键 词：动脉粥样硬化 TRPC5基因 巨噬细胞 凋亡 AKT信号通路 

分 类 号：R543.5[医药卫生—心血管疾病]