山羊乳腺上皮细胞氧化损伤模型的建立

doi:10.16431/j.cnki.1671-7236.2022.05.032

Abstract

Abstract: 【Objective】 This experiment was conducted to investigate the optimal conditions of oxidative damage induced by H₂O₂ in goat mammary epithelial cell (GMEC),and construct a reliable and stable oxidative damage model,in order to provide model conditions for exploring the protective mechanism of anthocyanins on oxidative damage in Dioscorea alata L.【Method】 In this experiment,two factors complete random design was used,the first factor was the concentration of H₂O₂ (0 (control group),350,430,460,490,520 and 550 μmol/L),and the second factor was the treatment time of GMEC (8,11,14 and 17 h),respectively.Treatment groups with cell survival rate lower than 60% were discarded,and treatment concentration and time were readjusted according to the results.Subsequently,the activity of glutathione peroxidase (GSH-Px),the content of reactive oxygen species (ROS) and malondialdehyde (MDA) in cells,the activity of lactate dehydrogenase (LDH),catalase (CAT),superoxide dismutase (SOD) and the total antioxidant capacity (T-AOC) in culture medium were measured by the kit,so as to determine the best treatment mode of H₂O₂.【Result】 Compared with control group,the survival rate of GMEC in all treatment groups were significantly decreased after 8 h treatment (P<0.05).The survival rate of GMEC in 360,430 and 460 μmol/L treatment groups were reduced to 64.6%,72.9% and 66.2% after 8 h treatment,which basically met the screening requirement of cell survival rate (70%).Therefore,350,430,460,490 and 520 μmol/L H₂O₂ treatments were selected for 4,6,8,10 h for follow-up tests.After adjustment, the LDH activity in all treatment groups was significantly higher than that of control group at different treatment times (P<0.05), and gradually increased with the extension of time, except 430 μmol/L treatment group for 4 h. The LDH activity of all treatment groups reached the maximum value at 10 h, and was proportional to the concentration. Compared with control group, ROS content in all treatment groups at 6 h was significantly increased (P<0.05), and ROS content in all treatment groups at 8 h was significantly increased compared with 6 h (P<0.05), indicating that 8 h was the turning point of ROS content change. Compared with control group, MDA content in 430, 490 and 520 μmol/L treatment groups was significantly increased after 8 h treatment (P<0.05), and MDA content in 430 μmol/L treatment group was higher than that of control group (P<0.05). The activities of CAT, SOD, GSH-Px and T-AOC in all groups were decreased to varying degrees from 6 to 8 h after treatment, and only the activities of CAT, SOD, GSH-Px and T-AOC in 430 μmol/L treatment group were significantly lower than that of control group at 8 h (P<0.05).【Conclusion】 Therefore,the method that GMEC treated with 430 μmol/L H₂O₂ for 8 h met the need of constructing oxidative damage model.

Key words: hydrogen peroxide; goat mammary epithelial cells; oxidative damage; survival rate; antioxidant

CLC Number:

S858.27

ZHANG Yuanxin, WANG Yanyu, YUN Yanhong, CHEN Junpu, SHI Huiyu, WANG Xuemei. Establishment of Oxidative Damage Model of Goat Mammary Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1917-1925.

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Establishment of Oxidative Damage Model of Goat Mammary Epithelial Cells

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