Background: Ischemic stroke (IS) is a serious cerebrovascular disease. Excessive levels of acetylated proteins are related to neuronal resistance to ischemia. Therefore, it is crucial to screen for histone acetylation regulatory-related biomarkers for IS. Methods: To identify differentially expressed genes (DEGs) in GSE16561, we executed a differential expression analysis (normal versus IS). Meanwhile, key module genes connected with single-sample gene set enrichment analysis (ssGSEA) enrichment score of histone acetylation regulatory related genes (HARGs) were identified by weighted correlation network analysis (WGCNA). Then, histone acetylation regulatory-related DEGs (HAR-DEGs) were attained by overlapping DEGs and key module genes. Three machine learning algorithms, expression and receiver operating characteristic (ROC) analysis, were employed to screen histone acetylation for regulatory related biomarkers. Afterwards, we carried out functional enrichment and immune microenvironment analysis on biomarkers.The disease association of biomarkers was analyzed. Finally, expression levels of biomarkers were assessed using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results:A total of 550 DEGs and 137 key module genes related to ssGSEA enrichment scores of HARGs were added up respectively. Through intersection of DEGs and key module genes, 44 HAR-DEGs were selected. Moreover, CREBBP and CKAP4 were identified as histone acetylation regulatory-related biomarkers in IS. We found that both biomarkers were linked with some immune-related signaling pathways, such as complement and coagulation cascade pathways. In addition, CREBBP was inversely linked with score of CD8 + T cells, while CKAP4 was positively correla t ed with score of M0 macrophages. Both biomarkers were found to be associated with brain injury and disease. Finally, in line with the outcomes of transcriptome data analysis, expression of CREBBP and CKAP4 was significantly elevated in clinical IS sample compared to versus control samples. Conclusion:In summary, we obtained two biomarkers ( CREBBP and CKAP4) that are connected with regulation of histone acetylation in relation to IS. These findings may laya theoretical basis for treatment of IS.