Bioinformatics Analysis And Validation Of Ferroptosis-related Genes In Ischemic Cardiomyopathy

Background: Ischemic cardiomyopathy(ICM)is a disorder that results from a persistent imbalance between the heart’s oxygen supply and demand.As the illness worsens,cardiomyocytes are gradually lost,which causes myocardial scarring and ventricular failure.The pathophysiology of ICM comprises a variety of metabolic,neurohumoral,and inflammatory alterations including cardiomyocyte hypertrophy,fibrosis,inflammation,oxidative stress,aberrant calcium metabolism,and cell death.ICM may be significantly impacted by ferroptosis,a kind of iron-dependent cell death regulation different from apoptosis and necrosis.In order to investigate the potential function of ferroptosis in ICM,the expression of genes associated to ferroptosis in ICM was screened and examined in this work.Objective: To inquire into the biological function of genes associated with ferroptosis and how they are expressed in ICM.Method:(1)Screening and classification of differentially expressed genes in ICM;(2)Co-screening and WGCNA of differentially expressed genes and ferroptosis-related genes;(3)Build protein-protein interaction networks;(4)GO analysis and KEGG analysis of differentially expressed ferroptosis-related genes in ICM;(5)ROC validation of hub genes;(6)Differential expression of hub genes was verified by CCK-8 experiments,Quantitative real-time reverse transcription PCR(q RT-PCR),and Western blotting analysis;(7)Drug prediction for hub genes.Result:(1)A total of 19 DEFRGs were identified through bioinformatics analysis,and GO and KEGG analysis revealed that these genes’ primary functions involved protein phosphorylation,regulation of enzyme activity,and conditional smooth muscle cell proliferation.They may also have been involved in the HIF-1signaling pathway;(2)The top 10 hub genes were generated by building protein-protein interaction networks and using the MCC algorithm.After performing ROC verification,7 hub genes with effective diagnostic performance were selected.;(3)In the hypoxia / reoxygenation model,q PCR experiments revealed that the expression of SLC40A1,SNCA,and GJA1 was significantly different from that in the control group,and the difference could be suppressed by the ferroptosis inhibitor Fer-1;(4)The SLC40A1,SNCA,and GJA1 genes may be affected therapeutically by tretinoin,according to the DSig DB database.Conclusion: We found three hub genes in this study — SLC40A1,SNCA,and GJA1 — through bioinformatics analysis of the microarray dataset GSE26887.We then confirmed through experiments that the aforementioned genes are indeed expressed differently in ICM and can be inhibited by the ferroptosis inhibitor Fer-1,demonstrating that the aforementioned genes may become potential therapeutic targets in the pathophysiological process of ICM.