| 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24 h，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。 |
| 关键词: 牛疱疹病毒Ⅰ型 MDBK细胞 细胞凋亡 |
| DOI： |
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| 基金项目: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） |
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| Preliminary study of apoptosis of MDBK cells induced by BHV-Ⅰ |
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ZHANG Kuan, XU Xinggang, TONG Dewen
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College of Veterinary Medicine ,Northwest A&F University
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| Abstract: |
| 【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. |
| Key words: bovine herpes virus-Ⅰ MDBK cell apoptosis |
