热应激诱导奶牛乳腺上皮细胞凋亡及其对HSP70基因表达的影响

河南农业科学 ›› 2017, Vol. 46 ›› Issue (8): 142-146.

热应激诱导奶牛乳腺上皮细胞凋亡及其对HSP70基因表达的影响

张响英^1,2，唐现文¹，陈静波¹，李腾腾¹

1.江苏农牧科技职业学院，江苏泰州 225300； 2.南京农业大学，江苏南京 210095

收稿日期:2017-02-26 出版日期:2017-08-15 发布日期:2017-08-15
作者简介:张响英（1974-），女，山东烟台人，副教授，博士，主要从事动物遗传繁育与生物技术研究。E-mail:724673081@qq.com
基金资助:
江苏省科技支撑计划项目(BE20083062)；江苏农牧科技职业学院大学生创新创业项目（201612806053P）

Received:2017-02-26 Published:2017-08-15 Online:2017-08-15

摘要/Abstract

摘要： 为探讨热应激对乳腺上皮细胞凋亡的影响，以体外培养的奶牛乳腺上皮细胞为模型，取对数生长期的乳腺上皮细胞分别置于39 ℃和41 ℃热处理1 h（以37 ℃正常培养的细胞为对照），在37 ℃恢复培养0、6、12 h后收集细胞，采用MTT法检测细胞生长抑制率，Annexin V/PI 双染法检测细胞凋亡，比色法分析Caspase-3活性，荧光定量PCR检测HSP70基因的表达水平。结果表明，39 ℃热应激组细胞生长抑制率和死亡率高峰期均发生在热修复6 h，细胞死亡率为15.42%，而41 ℃热应激组发生在热修复12 h，细胞死亡率高达23.18%；高温处理后，Caspase-3活性呈上升趋势；39 ℃热应激组在热恢复6 h，HSP70基因表达量显著高于对照组，41 ℃热应激组在热恢复0、6、12 h，HSP70基因的表达量分别为对照组的3.53倍、5.61倍和2.93倍。综上可知，高温应激激活了Caspase-3活性，从而诱导乳腺上皮细胞凋亡，并随高温强度的增加而作用增强，同时高温诱导了HSP70基因过表达，协助细胞获得热耐受性。

关键词: 乳腺上皮细胞； , 热应激； , 细胞凋亡； , HSP70基因

张响英，唐现文，陈静波，李腾腾. 热应激诱导奶牛乳腺上皮细胞凋亡及其对HSP70基因表达的影响[J]. 河南农业科学, 2017, 46(8): 142-146.

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