Protective Mechanism of Dihuang Yizhi Formula Against H2O2-Induced Injury in PC12 Cells
Objective:To investigate the protective effects of Dihuang Yizhi Formula against oxidative damage induced by H2O2 in PC12 cells,and to explore its potential protective mechanisms through proteomics.Methods:PC 12 cells were pretreated with different concentrations of Dihuang Yizhi Formula and then subjected to oxidative stress damage induced by H2O2.The levels of malondialdehyde(MDA)and 8-hydroxydeoxyguanosine(8-OHdG),as well as the activities of superoxide dismutase(SOD),glutathione reductase(GR),and glutathione peroxidase(GSH-Px),were measured using kits.For PC12 cells treated with 100 mg/mL Dihuang Yizhi Formula and subjected to H2O2-induced damage,differential protein expression was screened through mass spectrometry,followed by GO enrichment analysis,KEGG enrichment analysis,and protein interaction network analysis.Results:Pretreatment with Dihuang Yizhi Formula improved the activity of antioxidant enzymes(SOD,GSH-Px,GR)and increased intracellular GSH levels in H2O2-damaged PC12 cells;it also reduced MDA and 8-OHdG levels and decreased cell apoptosis.Following treatment with Dihuang Yizhi Formula,58 differential proteins were identified in H2O2-damaged PC12 cells.Compared to the model group,18 proteins were upregulated and 40 proteins were downregulated in the traditional Chinese medicine group.These differential proteins are involved in regulating oxidative stress responses,protein phosphorylation,peptide responses,and modulating pathways related to ferroptosis and mitochondrial autophagy.Dihuang Yizhi Formula can improve oxidative stress-induced cellular damage by regulating proteins such as GPX4,GPX1,and Jun.Conclusion:Dihuang Yizhi Formula can alleviate oxidative stress damage induced by H2O2,and the identified differential proteins provide new targets for investigating the mechanisms of Dihuang Yizhi Formula in the prevention and treatment of Alzheimer's disease,particularly through the ferroptosis pathway.
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