This study aimed to investigate the free radical scavenging ability of diallyl disulfide(DADS)and its protective effect against hydrogen peroxide(H2O2)-induced oxidative stress in Hu sheep rumen epithelial cells(RECs).Hu sheep RECs were treated with different concentrations(0,50,100,150,200 and 250 μmol/L)of H2O2 and different concentrations(0,25,50,75,100,150 and 200 μmol/L)of DADS for 24 hours.Cell viability was assessed using the CCK-8 assay,and intracellular reactive oxygen species(ROS)content was measured using fluorescent probes.Appropriate concentrations of H2O2 for establishing an oxidative stress model in Hu sheep RECs and safe concentration range of DADS were determined.Subsequent-ly,the RECs were pretreated with different concentrations(0,25,75 and 150 μmol/L)of DADS for 2 hours,followed by treatment with 200 μmol/L H2O2 for 24 hours.Cell viability and intracellular ROS content were assessed.The total antioxidant capacity(T-AOC),the content of malondialdehyde(MDA),and the ac-tivities of catalase(CAT),superoxide dismutase(SOD)and glutathione peroxidase(GPX)in Hu sheep RECs were measured using assay kits.Real-time fluorescence quantitative PCR(qRT-PCR)was performed to evaluate the mRNA relative expression levels of SOD,CAT and GPX.The results showed as follows:com-pared with the blank control group(0 µmol/L H2O2),200 μmol/L H2O2 significantly decreased cell viability and significantly increased intracellular ROS content(P＜0.05),inducing oxidative stress in Hu sheep RECs.Compared with the blank control group(0 μmol/L DADS),DADS at concentrations ranging from 25 to 150 μmol/L did not significantly affect the cell viability(P＞0.05).Compared to the blank control group(0 μmol/L H2O2 and 0 µmol/L DADS),200 μmol/L H2O2 significantly decreased cell viability(P＜0.05),significantly increased intracellular ROS and MDA contents(P＜0.05),and significantly reduced intracellular T-AOC,the mRNA relative expression level and the activity of SOD(P＜0.05);compared with the oxidative stress model group(induced by 200 μmol/L H2O2),pretreatment of Hu sheep RECs with 75 and 150 µmol/L DADS significantly reduced intracellular ROS and MDA contents(P＜0.05),and significantly increased intracellular T-AOC,the mRNA relative expression level and the activity of SOD(P＜0.05).In con-clusion,appropriate concentrations of DADS can effectively alleviate H2O2-induced oxidative stress in Hu sheep RECs.[Chinese Journal of Animal Nutrition,2023,35(12):8036-8045]
Hu sheeprumen epithelial cellsdiallyl disulfideoxidative stress
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