Heat-induced apoptosis and gene expression in bovine mammary epithelial cells
Han Hu A B C , Jiaqi Wang A B C D , Haina Gao A B C , Songli Li A B C , Yangdong Zhang A B C and Nan Zheng A B C
+ Author Affiliations
- Author Affiliations
A Ministry of Agriculture – Milk Risk Assessment Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
B Ministry of Agriculture – Milk and Dairy Product Inspection Centre, Beijing 100193, China.
C State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
D Corresponding author. Email: wangjiaqinmqc@126.com
Animal Production Science 56(5) 918-926 https://doi.org/10.1071/AN14420
Submitted: 19 March 2014 Accepted: 20 October 2014 Published: 26 February 2015
Abstract
The objective of this study was to identify the apoptosis and cell-defence response of bovine mammary epithelial cells under heat stress (HS). Cells were exposed to either 38°C or 42°C for 0.5, 1, 3, 5, 8, or 12 h, and the transcription of heat shock proteins (Hsps), Bcl-2 family, caspases and apoptosis-regulated genes were quantified by quantitative real-time polymerase chain reaction. Caspase-3, -7 and -8 were markedly upregulated by HS and the peak gene abundance appeared at 5 h. However, the same family numbers, caspase-6 and -9 were sustained downregulated in HS. The expression of anti-apoptotic gene Bcl-2, Bcl-2A and Mcl-1 increased sharply in HS but returned to pre-HS levels after 8 h. The pro-apoptotic genes: Bax, Bak and Bid were downregulated during HS. The striking changes of signalling factors of apoptosis: tumour necrosis factor receptor, p53, Apaf-1 was upregulated, and Fas was downregulated in HS. Stress proteins Hsp genes (hsp27, hsp70 and hsp90) were generally increased at 42°C and this was especially apparent for hsp70 transcription as it was increased 14-fold at 1 h. Simultaneously, HS induced cell apoptosis, and the peak of apoptosis rate appeared at 3 and 5 h, which were assessed by flow cytometry. Our results suggest that HS induces cell apoptosis, disturbs the normal biological activity, and aroused intracellular thermotolerance responses of bovine mammary epithelial cells.
Additional keywords: bovine mammary epithelial cell, heat shock proteins, heat stress.
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