摘要
目的:探讨NOX1对TNF-α诱导的A549细胞凋亡的影响。方法:将A549细胞分为3组,空白组、TNF-α组和TNF-α+NOX1 siRNA组。TNF-α+NOX1 siRNA组细胞采用瞬时转染技术转染特异性NOX1 siRNA,转染12 h后用含TNF-α(10μg/L)的培养基继续培养;空白组和TNF-α组细胞分别用培养基和含TNF-α的培养基培养。48h后,采用Annexin V-FITC和PI双染法检测细胞凋亡率,DCFH荧光标记法检测细胞中ROS的表达量,Western blot法检测NOX1及p-JNK蛋白的表达水平。结果:与空白组相比,TNF-α组细胞凋亡率和ROS表达量升高(P<0.05),NOX1和p-JNK蛋白表达水平升高(P<0.05);与TNF-α组比较,TNF-α+NOX1 siRNA组细胞凋亡率和ROS表达量降低(P<0.05),细胞中NOX1和p-JNK蛋白表达水平也降低(P<0.05)。结论:NOX1可能通过增加ROS的表达,进而激活JNK/MAPK信号通路,引起A549细胞的凋亡。
Aim： To investigate the effect of NOX1 on apoptosis of A549 cells induced by TNF-α.Methods： A549 cells were allocated into 3 groups：blank control group,TNF-α group and TNF-α＋NOX1 siRNA group. Cells in TNF-α＋NOX1 siRNA group were transfected with NOX1 siRNA through the transient transfection technology for 12 h,then cultured with TNF-α（10 μg/L）for 48 h. Cells in blank control group and TNF-α group were only cultured with medium and TNF-α（10 μg/L）for 48 h,respectively.After culture, the apoptosis rate was detected by Annexin V-FITC and PI staining,the ROS level in cells was detected by DCFH fluorescent probe method, and the expressions of NOX1 and p-JNK protein were detected through Western blot.Results： Compared with blank control group,the apoptosis rate,ROS level and the expressions of NOX1 and p-JNK were increased in TNF-α group（P〈0.05）; while compared with TNF-α group, the apoptosis rate,ROS level and the expressions of NOX1 and p-JNK were decreased in TNF-α＋NOX1 siRNA group（P〈0.05）.Conclusion： NOX1 could increase ROS level, then active JNK/MAPK signal pathway, and induce A549 cell apoptosis.
作者
夏艳秋
王娜
阙菡雅
徐小艳
薛腾
燕贞
姚武
刘莹
周舫
XIA Yanqiu WANG Na QUE Hanya XU Xiaoyan XUE Teng YAN Zhen YAO Wu LIU Ying ZHOU Fang(Department of Occupational Health, College of Public Health ,Zhengzhou University,Zhengzhou 450001 Department of Public Health, Center for Disease Prevention and Control of Zhengzhou ,Zhengzhou 450000 Department of Respiratory Diseases ,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2016年第5期591-594,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省高等学校重点科研项目 16A330008
