牛疱疹病毒Ⅰ型诱导MDBK细胞凋亡的初步研究  

Preliminary study of apoptosis of MDBK cells induced by BHV-Ⅰ

作　　者：张宽[1] 许信刚[1] 童德文[1] 丁丽[1] 李兆才[1] 李伟[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2011年第12期21-25,30,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：教育部新世纪优秀人才支持计划项目(NCET-07-0701);西北农林科技大学青年学术骨干项目(E111020901)

摘　　要：【目的】研究牛疱疹病毒Ⅰ型(BHV-Ⅰ)对牛肾细胞(MDBK细胞)的作用,为探讨BHV-Ⅰ诱导细胞凋亡的分子机制奠定基础。【方法】常规方法培养MDBK细胞,于对数生长期感染BHV-Ⅰ,同时设立对照组,于感染后不同时间取样,采用细胞形态观察、细胞活性检测、细胞核形态观察、DNA ladder检测、细胞凋亡率检测和细胞内Caspase-3活性检测等方法,对BHV-Ⅰ感染的MDBK细胞进行凋亡指标检测。【结果】接毒后24h,MDBK细胞即出现明显的细胞病变。活性检测表明,感染BHV-Ⅰ的细胞活性降低,并且呈病毒感染量和感染时间的依赖性。感染细胞出现核固缩,并呈新月形变化。DNA电泳结果和流式细胞术检测结果均表明,感染BHV-Ⅰ的细胞DNA出现规律的片段化,细胞凋亡比例随感染时间延长而增加,细胞内Caspase-3活性逐渐升高。【结论】BHV-Ⅰ能够诱导MDBK细胞凋亡,并且在诱导细胞凋亡过程中有Caspase-3参与。【Objective】 The aim of this study was to investigate whether BHV-Ⅰ could cause MDBK cells apoptosis,which would lay theoretical foundation for the further observation of BHV-Ⅰ induced apoptosis.【Method】 MBDK cells were cultured and infected with BHV-Ⅰ.Subsequently,cells were collected at various time post infection,including cells of control group.Then AO-EB straining,flow cytometry and DNA ladder tests were used to detect the apoptotic MDBK cells.Besides,spectrophotometry was used to examine the activity of caspase-3.【Result】 24 h post infection,cytopathic effect appeared and vitality of cells began to decrease.Apoptosis appeared in MDBK cells infected with BHV-Ⅰwhich was judged by DNA laddering,Flow cytometry and chromatin condensation.With the increase of apoptotic cells,the level of caspase-3 rose.【Conclusion】 BHV-Ⅰ can induce MDBK cells to undergo apoptosis which involves caspases-3 activation.

关 键 词：牛疱疹病毒Ⅰ型 MDBK细胞 细胞凋亡 

分 类 号：S858.215.3[农业科学—临床兽医学]