连二亚硫酸钠诱导大鼠肝细胞氧化应激模型的建立

doi:10.16431/j.cnki.1671-7236.2023.06.005

Abstract

Abstract: 【Objective】 This study was aimed to explore the preparation method of rat hepatocyte (BRL-3A) oxidative stress model induced by sodium hydrosulfite (Na₂S₂O₄), in order to establish a stable oxidative stress model of BRL-3A cells.【Method】 The medium containing Na₂S₂O₄ was used to cause hypoxia of cells, and the normal medium was replaced to reoxygenate BRL-3A cells, thus causing oxidative stress damage of cells.BRL-3A cells were divided into 5 groups, and the control group was cultured normally in a cell incubator (0 mmol/L Na₂S₂O₄), the experiment group was treated with different concentrations (0.1, 1, 5 and 10 mmol/L) of Na₂S₂O₄ for 4 h of hypoxia and 2 h of reoxygenation, then the cell viabiliaty rate was detected by CCK-8 assay.The fluorescent probe DCFH-DA method was used to detect the release of reactive oxygen species (ROS) in cells.The activity of lactate dehydrogenase (LDH) in cell culture medium, the contents of malondialdehyde (MDA) and glutathione (GSH), and the activity of superoxide dismutase (SOD) in BRL-3A cells were detected by kit.【Result】 After hypoxia/reoxygenation treament, compared with blank control group, with the increase of Na₂S₂O₄ concentration, the viability rate of BRL-3A cells was gradually decreased (P<0.01), the release of ROS was gradually increased (P<0.01), the activity of LDH and the content of MDA were increased in a concentration dependent manner (P<0.01), the content of GSH and the activity of SOD were decreased in a concentration dependent manner (P<0.05 or P<0.01).【Conclusion】 This study showed that a stable oxidative stress model of BRL-3A cells could be established which was induced by 5 mmol/L Na₂S₂O₄ for hypoxia of 4 h and reoxygenation of 2 h.

Key words: BRL-3A cells; sodium hydrosulfite (Na₂S₂O₄); oxidative stress

CLC Number:

S852.21

SUN Chen, GE Sheng, NING Xiaqing, WANG Jiaqi, ZHANG Nuannuan, WANG Xiaoran, ZHANG Shixia. Oxidative Stress Model of BRL-3A Cells Induced by Sodium Hydrosulfite[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2217-2223.

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Oxidative Stress Model of BRL-3A Cells Induced by Sodium Hydrosulfite

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