Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Dinitrochlorobenzene Inhibiting Thioredoxin Reductase Activity

GONG Jian; YAN Sumei; SHI Binlin; GUO Xiaoyu; JIN Lu; GUO Yongmei

doi:10.3969/j.issn.1006-267x.2015.12.041

2015 , Vol. 27 >Issue 12: 3984 - 3990

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.12.041

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Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Dinitrochlorobenzene Inhibiting Thioredoxin Reductase Activity

GONG Jian ,
YAN Sumei ,
SHI Binlin ,
GUO Xiaoyu ,
JIN Lu ,
GUO Yongmei

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1. College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. College of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, China

Received date: 2015-07-02

Online published: 2015-12-14

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Abstract

Dinitrochlorobenzene(DNCB) was selected as the inhibitor of thioredoxin reductase(TrxR) activity to induce oxidative damage of bovine mammary epithelial cells(BMECs), and oxidative damage model was established by detecting cell proliferation rate, TrxR activity and antioxidant parameters in BMECs. The experiment was divided into two parts. The experiment 1 was conducted as a single factor randomized arrangement, and the 3th passage BMECs were divided into 35 groups with 8 replicates in each group. Cells were incubated with culture medium containing different concentrations of DNCB[0(control), 10, 30, 50, 100, 300 and 500μmol/L] for 2, 4, 6, 8 and 12 h to determine the appropriate action time of DNCB by detecting the cell proliferation rate. Based on the results in experiment 1, the experiment 2 was conducted as a single factor randomized arrangement, and the cells were divided into 7 groups with 6 replicates in each group. Cells were incubated with culture medium containing different concentrations of DNCB[0(control), 100, 200, 400, 600, 800 and 1000μmol/L] for 2 h to select the appropriate concentration of DNCB by measuring TrxR activity and antioxidant parameters. The results showed that compared with control group, the group of 300μmol/L DNCB for 2 h resulted in significant oxidative damage of BMECs, and cell proliferation rate reduced to 73.31%, TrxR activity decreased to 56.23%, the activities of glutathione peroxidase, superoxide dismutase and catalase also significantly decreased(P<0.05), and malondialdehyde content significantly increased(P<0.05). The results indicate that DNCB concentration of 300μmol/L and the time of 2 h can be selected as the appropriate action concentration and time to establish the oxidative damage model of BMECs.

Key words： bovine mammary epithelial cells; dinitrochlorobenzene; oxidative damage model

Cite this article

GONG Jian , YAN Sumei , SHI Binlin , GUO Xiaoyu , JIN Lu , GUO Yongmei . Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Dinitrochlorobenzene Inhibiting Thioredoxin Reductase Activity[J]. Chinese Journal of Animal Nutrition, 2015 , 27(12) : 3984 -3990 . DOI: 10.3969/j.issn.1006-267x.2015.12.041

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