冠心病中铁死亡和N6-甲基腺苷修饰的相关性研究

The correlation between ferroptosis and N6-methyladenosine modification in coronary heart disease

目的:探讨冠心病(CHD)患者N6-甲基腺苷(m6A)修饰与铁死亡相关基因(FAGs)的相关性。方法:检索基因表达综合(GEO)数据库中的GSE90076和GSE113079数据集,进行CHD组和对照组的差异表达基因(DEGs)分析。利用R语言对选定的DEGs执行基因本体(GO)、基因组百科全书(KEGG)和基因集变异分析(GSVA)。随后对其进行加权基因共表达网络分析(WGCNA),确定与铁死亡相关性最高的重要模块,进行GO和KEGG分析。再从DEGs中选出常规m6A修饰基因和FAGs,通过WGCNA中的数据,选出相关性最强的m6A修饰基因和FAGs。采用实时荧光定量PCR(qRT-PCR)和Western blotting(WB)方法对筛选出的相关基因进行验证。结果:在两个数据集中共同鉴定出2 159个DEGs, GO和KEGG结果表明,这些基因主要在mRNA修饰及代谢、内质网蛋白过程和炎症相关通路富集。GSVA方法发现DEGs主要富集在铁死亡等通路。WGCNA分析显示,黑色和黄绿色模块分别与铁死亡呈最强的正相关性和负相关性。最终筛选出了18个m6A相关基因和13个FAGs,分析这些基因在WGCNA中的数据,得到4组相关性强的基因,并在临床上收集CHD和正常人全血样本各10例进行了qRT-PCR和WB验证。结论:CHD患者m6A RNA甲基化与铁死亡之间存在显著的相关性。

Objective:To explore the correlation between N6-methyladenosine(m6A)modification and ferrop-tosis-associated genes(FAGs)in coronary heart disease(CHD).Methods:We performed differential expression genes(DEGs)analysis in the CHD group and the control group in the Gene Expression Omnibus(GEO)database using the GSE90076 and GSEl13079 datasets.The selected DEGs were subjected to Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),and Gene Set Variation Analysis(GSVA)using the R programming language.Subsequently,Weighted Gene Coexpression Network Analysis(WGCNA)was performed to identify the key module with the highest correlation to ferroptosis,followed by GO and KEGG analyses on these modules.Then,from the DEGs,conventional m6A modification genes and FAGs were selected,and the most highly corre-lated m6A modification genes and FAGs were screened out using the data from the WGCNA.The selected genes were validated using real-time fluorescence quantitative PCR(qRT-PCR)and Western blotting(WB)methods.Re-sults:In the two datasets,a total of 2159 DEGs were identified.The results of GO and KEGG analyses indicated that these genes were primarily enriched in pathways related to mRNA modification and metabolism,endoplasmic reticulum protein processes,and inflammatory pathways.The GSVA method revealed that the DEGs were pre-dominantly enriched in pathways such as ferroptosis.The WGCNA analysis revealed that the black and greenyel-low modules were most strongly positively and negatively correlated with ferroptosis,respectively.Eighteen m6A-related genes and thirteen FAGs were screened out.By analyzing the data of these genes in WGCNA,four groups of highly correlated genes were obtained.Subsequently,1o full blood samples each from CHD patients and normal individuals were collected for validation through qRT-PCR and WB. Conclusion: There is a significant correlationbetween m6A RNA methylation and ferroptosis in CHD.