热应激对体外培养奶牛乳腺上皮细胞生长、凋亡及其热休克蛋白mRNA转录的影响

畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (5): 600-607.doi:

热应激对体外培养奶牛乳腺上皮细胞生长、凋亡及其热休克蛋白mRNA转录的影响

周振峰^1,2,，崔瑞莲²，王加启²*，魏宏阳²，卜登攀²，胡菡^1,2,，周凌云²

1.甘肃农业大学动物科技学院，兰州 730070；2.中国农业科学院北京畜牧兽医研究所动物营养学国家重点实验室，北京 100193

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-05-25 发布日期:2010-05-25
通讯作者: 王加启

Cell Growth, Apoptosis and the mRNA Transcription of Heat Shock Protein:Effects of Heat Stress on Bovine Mammary Epithelial Cells

ZHOU Zhen-feng^1,2, CUI Rui-lian², WANG Jia-qi²*, WEI Hong-yang²,
BU Deng-pan², HU Han^1,2,, ZHOU Ling-yun²

1. Faculty of Animal Science & Technology, Gansu Agricultural University,Lanzhou 730070, China; 2.State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:1900-01-01 Revised:1900-01-01 Online:2010-05-25 Published:2010-05-25

摘要/Abstract

摘要： 本研究旨在探讨热应激对奶牛乳腺上皮细胞生长、凋亡及热休克蛋白（Heat shock protein，HSP）mRNA转录的影响。本文以体外高温(42 ℃)作为热应激模型温度，38 ℃作为对照温度培养奶牛乳腺上皮细胞，并采用台盼蓝染色绘制生长曲线，细胞流式检测细胞凋亡及RT-qPCR检测HSPs mRNA转录。结果表明， 42 ℃高温处理使得奶牛乳腺上皮细胞生长停滞，细胞数量在培养的第2天已显著少于对照组(38 ℃)（P<0.05），第3～7天极显著的少于对照组 (P<0.01)；细胞凋亡率在42 ℃高温培养测定的第3小时达到最大值；42 ℃高温使得乳腺上皮细胞热休克蛋白HSP27、70、90和热休克因子HSF-1（Heat shock transcriptional factor)的mRNA转录水平上调，分别为对照组的2.72、7.48、2.69和2.71倍，与对照组相比均差异极显著（P<0.01）。结果提示，高温抑制乳腺上皮细胞正常生长，促使细胞凋亡，并诱导乳腺上皮细胞HSF-1和HSPs mRNA的转录表达。

Abstract: The experiment was conducted to study the effects of heat stress on bovine mammary epithelial cells in vitro. Bovine mammary epithelial cells were exposed to 42 ℃ as the heat stress model and 38 ℃ as control, then cell growth, apoptosis and heat shock protein expression were detected by trypan blue dying, flow cytometry FCM , RT-qPCR and ELISA. The results showed as follows: cell growth stagnated at 42 ℃; the cell number decreased markedly (P<0.05) than control 38 ℃ at the second day, and decreased extremely significantly (P<0.01) from the third to the seventh day; the apoptosis rate was the highest at the 3rd hour after heat treatment; the mRNA expression level of heat shock protein 27, 70, 90 (HSP27,HSP70,HSP90) and heat shock factor-1 (HSF-1) were 2.72, 7.48, 2.69 and 2.71 folds as control group, separately, which were all significantly higher than control(P<0.01). In conclusion, heat stress could inhibit cell natural growth, induce apoptosis, and upregulate the mRNA of HSPs.

周振峰;;崔瑞莲;王加启;魏宏阳;卜登攀;胡菡;;周凌云. 热应激对体外培养奶牛乳腺上皮细胞生长、凋亡及其热休克蛋白mRNA转录的影响[J]. 畜牧兽医学报, 2010, 41(5): 600-607.

ZHOU Zhen-feng;CUI Rui-lian;WANG Jia-qi;WEI Hong-yang;BU Deng-pan;HU Han;;ZHOU Ling-yun. Cell Growth, Apoptosis and the mRNA Transcription of Heat Shock Protein:Effects of Heat Stress on Bovine Mammary Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(5): 600-607.

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