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Effects of glutamine on the growth, proliferation and expression of HSP70 in bovine mammary epithelial cells#br#

  


  1. (1 College of Animal Science and Technology, Nanjing Agricultural University, Jiangsu 210095, China; 2 Jiangsu Agri\|animal Husbandry Vocational College, Taizhou 225300, China)
  • Online:2015-09-25 Published:2015-09-23

Abstract: The present study was performed to investigate the effects of glutamine on the growth, proliferation and expression of HSP70 gene in bovine mammary epithelial cells. Bovine mammary epithelial cells in logarithmic growth phase were used for test. The cells were cultured in DMEM with different concentrations (0, 05, 10, 20, 40, 80 and 160 mmol·L-1) of glutamine for 24 h, then cells and supernatant were collected. The cells were cultured in DMEM with 8 mmol·L-1 glutamine at 37℃ for 0, 3, 6, 12, 24, 48, 72 h,respectively, then cells and supernatant were collected. The survival rate of cells was detected by MTT assay, LDH activity was detected by colorimetric assay, and the expression levels of HSF\|1 and HSP70 were tested by SYBR GreenⅠFQ\|PCR. The results showed that glutamine promoted the growth and proliferation of bovine mammary epithelial cells. The survival rate of cells was the highest at the glutamine concentration of 20 mmol·L-1, and it maintained higher level at the glutamine concentration of 40-80 mmol·L-1. The expression trend of HSP70 was the same with HSF\|1 and both reached the peak with the glutamine concentration of 80 mmol·L-1 at 24 h. Compared with the optimal dosage of 20 mmol·L-1, it was 849 (P<001) and 590 times (P<001). These results suggested that glutamine could improve the growth and proliferation of bovine mammary epithelial cells in vitro, and it could up\|regulate the expression of HSP70 through promoting the transcriptional activity of HSF\|1 mRNA.

Key words: glutamine, mammary epithelial cell, HSP70, bovine