Objective:To investigate the protective role of L-isoleucine in alleviating oxidative stress induced by H202 in porcine sertoli cells,to determine the optimal concentration of L-isoleucine for mitigating the decline in cell viability due to oxidative stress,and to assess,the effects on cell proliferation,apoptosis,and antioxidant levels in porcine Sertoli cells at the optinal concentranon.Methods:Cell viability was examined by using the CCK-8 assay to select the most suitable concentration of L-isoleucine for protecting porcine Sertoli cells under oxidative stress conditions.Flow cytometry was utilized to detect the cell cycle and apoptosis of Sertoli cells at the optimal L-isoleucine concentration.The levels of Superoxide dismutase(T-SOD),Total antioxidant capacity(T-AOC),Glutathione peroxidase(GSH-Px),and Malondialdehyde(MDA)were measured by using specific assay kits.Results:Compared to the oxidative stress alanine nitrogen control group and other concentrations of oxidative stress isoleucine groups,cell proliferation vitality was significantly elevated in the 32 mmol/L oxidative stress isoleucine group.Relative to the oxidative stress untreated group and the oxidative stress alanine nitrogen control group,the oxidative stress isoleucine group exhibited a comparatively lower ratio of cells in the G0/G1 phase,with no notable differences in the proportions of the S and G2/M phases.In comparison to the oxidative stress untreated group,the proportion of cell apoptosis was significantly reduced in the oxidative stress isoleucine-treated group,with no significant difference when compared to the oxidative stress nitrogen control group.Isoleucine markedly increased T-SOD activity in comparison to both the non-oxidative stress alanine nitrogen control group and the oxidative stress alanine control group.It also significantly enhanced T-AOC levels in the oxidative stress isoleucine group,though it did not have a significant effect on GSH-Px and MDA.Conclusion:Under the experimental conditions of this study,32 mmol/L of isoleucine was found to be the most effective concentration,successfully mitigating the decline in cell viability,cell cycle dysregulation,and cell apoptosis caused by oxidative stress.
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