缺血性脑卒中相关的线粒体自噬和铁死亡关键基因筛选及实验验证*

doi:10.3870/j.issn.1672-0741.25.07.042

摘要/Abstract

摘要： 目的探讨缺血性脑卒中(ischemic stroke,IS)中线粒体自噬和铁死亡相关的关键基因,研究线粒体自噬、铁死亡与IS的关系。方法利用基因表达综合数据库、GeneCards数据库和铁死亡相关数据库分别获取IS基因表达谱、线粒体自噬和铁死亡相关基因。筛选出共同差异表达基因,并对其进行功能富集分析和蛋白-蛋白相互作用网络分析,结合2种拓扑算法分析获得候选基因。以数据集GSE137482对候选基因进行基因表达验证,经受试者工作特征(ROC)曲线评估诊断特征的准确性,得到最终靶点基因,最后通过数据集GSE61616和动物实验验证靶点基因。将SD大鼠随机分为假手术组和模型组,每组10只。运用蛋白质免疫印迹技术和实时荧光定量PCR检测靶点基因的表达水平。结果共筛选出36个共同差异表达基因,经GO和KEGG富集分析,主要参与自噬和铁死亡相关信号通路。选取SIRT3、STAT3、HMOX1和NFE2L2为最终靶点,其中SIRT3在IS中表达明显下降(P＜0.05),STAT3、HMOX1和NFE2L2表达升高(均P＜0.05),且在ROC曲线分析中AUC值均为1。动物实验中,IS模型组STAT3、HMOX1和NFE2L2的蛋白表达量上升,SIRT3表达下降(均P＜0.05)。结论 SIRT3、STAT3、HMOX1和NFE2L2可能是IS中参与线粒体自噬和铁死亡的关键共同基因。

关键词: 缺血性脑卒中, 线粒体自噬, 生物信息学, 铁死亡, 活性氧

Abstract: Objective To investigate key genes associated with mitophagy and ferroptosis in ischemic stroke(IS)and explore their relationship. Methods Gene expression profiles related to IS were obtained from the Gene Expression Omnibus(GEO),GeneCards,and ferroptosis-related databases.Commonly differentially expressed genes(co-DEGs)were identified,subjecting to functional enrichment and protein-protein interaction(PPI)network analyses.Topological algorithms were applied to screen potential candidate genes,whose diagnostic performance was evaluated by receiver operating characteristic(ROC)curve analysis.The identified target genes were further validated using independent disease-related datasets and animal experiments.Sprague-Dawley (SD)rats were randomly divided into sham(n=10)and model(n=10)groups.Western blotting and quantitative real-time PCR(qRT-PCR)were performed to assess gene expression levels. Results A total of 36 shared genes were identified,which were significantly enriched in pathways associated with autophagy and ferroptosis.Among them,SIRT3,STAT3,HMOX1,and NFE2L2 were identified as key targets.In IS,SIRT3 expression was markedly decreased(P<0.05),whereas STAT3,HMOX1,and NFE2L2 were significantly upregulated(P<0.05).ROC curve analysis yielded area under the curve(AUC)values of 1.0 for all four target genes.Consistent with the bioinformatics findings,animal experiments confirmed increased protein levels of STAT3,HMOX1,and NFE2L2,and decreased SIRT3 in the model group(P<0.05). Conclusion SIRT3,STAT3,HMOX1,and NFE2L2 may serve as critical genes linking mitophagy,ferroptosis,and IS pathogenesis.

Key words: ischemic stroke, mitophagy, bioinformatics, ferroptosis, reactive oxygen species

中图分类号:

R743.3

黄梦玲, 李瑞青, 黄金, 张丽红, 顾昌宇, 李晶晶. 缺血性脑卒中相关的线粒体自噬和铁死亡关键基因筛选及实验验证^*[J]. 华中科技大学学报（医学版）, 2026, 55(2): 164-174.

Huang Mengling, Li Ruiqing, Huang Jin et al. Bioinformatics-based Analysis of Key Genes Linking Mitophagy and Feroptosis in Ischemic Stroke[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2026, 55(2): 164-174.

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缺血性脑卒中相关的线粒体自噬和铁死亡关键基因筛选及实验验证^*

Bioinformatics-based Analysis of Key Genes Linking Mitophagy and Feroptosis in Ischemic Stroke

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