摘要
为探讨犬细小病毒(CPV)引起宿主细胞凋亡的分子机制,用CPV感染MDCK细胞,感染后通过台盼蓝染色法检测不同时间的细胞存活率,用Annexin V-FITC/PI法检测磷脂酰丝氨酸外翻情况,用试剂盒检测Caspase-3活性,分析CPV诱导细胞的凋亡。并通过流式细胞术检测细胞线粒体的损伤和ROS的产生探讨其分子机制。结果显示,随着病毒感染时间的不断延长,CPV可明显降低MDCK细胞存活率,促进磷脂酰丝氨酸外翻,增强Caspase-3活性,表明CPV可引起细胞凋亡。同时还发现CPV可明显提高细胞内ROS的水平,引起线粒体的损伤,这可能是CPV诱导细胞凋亡的重要分子基础。由此可见,CPV诱导MDCK细胞凋亡与CPV增加细胞ROS的产生和线粒体的损伤有关。
To investigate the molecular mechanism of canine parvovirus(CPV)-induced apoptosis of host cells,the MDCK cells were infected by CPV, and the cell viability was detected by Trypan blue staining at different time points. The apoptosis of the cells were detected by Annexin V-FITC/PI double staining for phosphatidylserine(PS) externalization on membrane and Caspase-3 activity was detected using a kit. To explore the mechanism,the mitochondrial damage and ROS production were detected by Flow cytometry. The results showed that CPV could decrease the survival rate of MDCK cells. The PS on outside of membrane was detected and the activity of caspase-3 was increased, which indicated that CPV could induce MDCK cell apoptosis. It was also found that CPV could significantly increase ROS production and mitochondrial damage, which may be the molecular basis for CPV-induced apoptosis. Therefore, it can be concluded that CPV-induced ROS production and mitochondrial damage are involved in CPV-induced MDCK cell apoptosis.
作者
王岩
张建楼
赵轶男
王晓敏
王猛
李笑然
仲飞
WANG Yan;ZHANG Jianlou;ZHAO Yinan;WANG Xiaomin;WANG Meng;LI Xiaoran;ZHONG Fei(College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001,China)
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2019年第3期99-103,114,共6页
Journal of Hebei Agricultural University
基金
河北农业大学大学生创新创业训练计划项目(201810086034)
河北省高等学校科学技术研究项目(QN2016189)
“十三五”国家重点研发计划项目(2016YFD0501002)
关键词
犬细小病毒
细胞凋亡
MDCK
ROS
线粒体损伤
canine parvovirus
apoptosis
MDCK cell
reactive oxygen species
mitochondria damage
