Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Hydrogen Peroxide

JIN Lu; YAN Sumei; SHI Binlin; SHI Huiyu; GUO Xiaoyu; LI Junliang

doi:10.3969/j.issn.1006-267x.2014.12.014

Chinese Journal of Animal Nutrition >

2014 , Vol. 26 >Issue 12: 3651 - 3658

DOI: https://doi.org/10.3969/j.issn.1006-267x.2014.12.014

Ruminant Nutrition

Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Hydrogen Peroxide

JIN Lu ,
YAN Sumei ,
SHI Binlin ,
SHI Huiyu ,
GUO Xiaoyu ,
LI Junliang

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College of Animal Science, Inner Mongolia Agricultural University, Huhhot 010018, China

Received date: 2014-06-23

Online published: 2014-12-05

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Abstract

This study was conducted to establish the oxidative damage model of bovine mammary epithelial cells (BMEC) using hydrogen peroxide (H₂O₂) as stress source and using survival rate and antioxidant parameters as judgment indices. Experiment 1 was conducted as a single factor randomized arrangement, and the 3rd passage BMEC were divided into 42 groups with 10 replicates in each group. Cells were incubated in different concentrations of H₂O₂ (0, 100, 200, 400, 600, 800 and 1 000 μmol/L,respectively) for 2, 4, 6, 8, 12 and 24 h, respectively. The cell count was determined, and the survival rate was calculated. On the basis of experiment 1 (optimal action time was 6 h), experiment 2 was also conducted as a single factor randomized arrangement, and the 3th passage BMEC were divided into 7 groups with 6 replicates in each group. Culture mediums containing different concentrations of H₂O₂ (0, 100, 200, 400, 600, 800 and 1 000 μmol/L, respectively) were used to act cell for 6 h. The cells and culture mediums were collected for the analysis of antioxidant parameters to select the appropriate concentration of H₂O₂ ensuring oxidative damage to cells. The results showed as follows: experiment 1, the BMEC survival rate reduced to 79.79% when the cells were induced by 600 μmol/L H₂O₂ for 6 h. Experiment 2, cell induced by 600 to 1 000 μmol/L H₂O₂ significantly decreased the activities of glutathione peroxidase, superoxide dismutase and catalase (P<0.05), and significantly increased malondialdehyde content compared with the other groups (P<0.05), but the differences among 600, 800 and 1 000 μmol/L groups were not significant (P>0.10). The condition that 600 μmol/L H₂O₂ reacted for 6 h is optimal for establishment of oxidative damage model in BMEC.

Key words： bovine mammary epithelial cells; hydrogen peroxide; oxidative damage model

Cite this article

JIN Lu , YAN Sumei , SHI Binlin , SHI Huiyu , GUO Xiaoyu , LI Junliang . Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Hydrogen Peroxide[J]. Chinese Journal of Animal Nutrition, 2014 , 26(12) : 3651 -3658 . DOI: 10.3969/j.issn.1006-267x.2014.12.014

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