Naringenin Ameliorates STZ-induced Oxidative Damage In SH-SY5Y Cells And Regulates Nrf2 And MTOR Expression

Objective: The intent of our study was to explore the mechanismof naringenin in reducing the oxidative damage induced by STZ by regulating Nrf2 and mTOR signaling pathways.Methods: CCK-8 was used to evaluate STZ treatment,STZ co-treatment with naringenin,and normal cultured SH-SY5 Y cells to assess cell viability;qPCR was used to detect the expression of gene;Evaluation of protein levels by Western blotting;flow cytometry Detect the cell apoptosis rate;Ed U proliferation staining suggested that the cells in Control group,STZ group,and STZ+Nar group all showed proliferation activity,but the cell density in STZ group was the lowest,and the cell density in STZ+Nar group was significantly higher than that in STZ group,inferior to Control group.The results of reactive oxygen staining showed that the STZ group had the most abundant ROS production,which was higher than that of the positive control.The ROS production of cells in the STZ+Nar group was significantly lower than that of the STZ group,and slightly higher than that of the Control group.TUNEL apoptosis staining showed that the control group had the least apoptosis,the STZ group had serious apoptosis,and most of the cells were shed.The STZ+Nar group had a high degree of apoptosis but Cells adhered well,indicating that naringenin played a protective role;Knockdown expression of Nrf2 and mTOR with siRNA and subsequent detection of gene expression and related proteins.Results: In this study,the CCK-8 test results implied that the higher concentration of STZ reduced the cell viability of SH-SY5 Y cells more significantly;this experiment also found 200 μ M naringenin It can improve the oxidative stress response induced by 8mM STZ.The results of fluorescent staining also provide evidence for our conclusion fromthe morphological aspect.Further,Western blotting was used to detect signaling pathway-related proteins,and it was found that naringenin regulates the expression of downstreamrelated proteins by enhancing Nrf2 expression and mTOR expression.Reverse transcriptase real-time quantitative PCR method proved that naringenin up-regulated the expression of Nrf2 and mTOR at the gene level.After knocking down Nrf2 by siRNA,it was found that it significantly affected the expression of mTOR pathway-related genes and proteins.Similarly,knocking down mTOR expression with siRNA affected the expression of Nrf2-related genes and proteins.Conclusion: The results of this experimental study indicate thatnaringenin can enhance the anti-oxidative stress capacity of SH-SY5 Y cells by enhancing Nrf2 and regulating the expression of mTOR,and improves the oxidative stress damage caused by STZ.