地黄益智方保护H_(2)O_(2)诱导的PC12细胞损伤的作用机制  

Protective Mechanism of Dihuang Yizhi Formula Against H_(2)O_(2)-Induced Injury in PC12 Cells

作　　者：张彤[1] 安红梅[1] 张立敏[1] ZHANG Tong;AN Hongmei;ZHANG Limin(Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032

出　　处：《中医药学报》2025年第1期20-28,共9页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(82104744)。

摘　　要：目的:研究地黄益智方对阿尔茨海默病(AD)中H_(2)O_(2)诱导的PC12细胞氧化损伤的保护作用,并通过蛋白质组学探讨其对PC12细胞的潜在保护机制。方法:不同浓度的地黄益智方预处理PC12细胞,然后用H_(2)O_(2)诱导氧化应激损伤。试剂盒检测丙二醛(MDA)、8-羟基脱氧鸟苷(8-OHdG)含量,以及超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GR)、谷胱甘肽过氧化物酶(GSH-Px)活性。100 mg/mL的地黄益智方处理H_(2)O_(2)损伤的PC12细胞,质谱分析筛选差异表达蛋白,并对其进行GO富集分析、KEGG富集分析和蛋白互作网络分析。结果:地黄益智方预处理PC12细胞可改善H_(2)O_(2)损伤诱导的SOD、GSH-Px、GR抗氧化酶活性和细胞内GSH的水平降低(P<0.05,P<0.01);降低MDA、8-OHdG含量及细胞凋亡(P<0.05,P<0.01)。地黄益智方作用后,H_(2)O_(2)损伤的PC12细胞有58个差异蛋白表达。相较于模型组,地黄益智方组中有18个蛋白表达发生上调,40个蛋白表达发生下调。差异蛋白涉及调节氧化应激反应、调节蛋白质磷酸化和调节肽反应等生物过程及调控细胞铁死亡通路、线粒体自噬通路等信号通路的表达。地黄益智方可通过调节GPX4、GPX1、Jun等蛋白改善氧化应激诱导的细胞损伤。结论:地黄益智方可以减轻H_(2)O_(2)诱导的氧化应激损伤,筛选出的差异蛋白为探究地黄益智方防治AD的作用机制提供了新的靶点——铁死亡通路。Objective:To investigate the protective effects of Dihuang Yizhi Formula against oxidative damage induced by H_(2)O_(2)in PC12 cells,and to explore its potential protective mechanisms through proteomics.Methods:PC12 cells were pretreated with different concentrations of Dihuang Yizhi Formula and then subjected to oxidative stress damage induced by H_(2)O_(2).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 H_(2)O_(2)-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 H_(2)O_(2)-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 H_(2)O_(2)-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 H_(2)O_(2),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.

关 键 词：地黄益智方 阿尔茨海默病 PC12 氧化应激 蛋白质组学 

分 类 号：R285.5[医药卫生—中药学]