实用心电与临床诊疗2025，Vol.34Issue(4)：481-493,13.DOI:10.13308/j.issn.2097-5716.2025.04.003
circHECTD1在脂多糖诱导巨噬细胞炎症反应中的N6-甲基腺苷依赖性调控机制
N6-methyladenosine-dependent regulatory mechanism of circHECTD1 in lipopolysaccharide-induced macrophage inflammatory response
摘要
Abstract
Objective To investigate the regulatory role and underlying mechanism of circular RNA circHECTD1,a potential diagnostic biomarker for coronary heart disease(CHD),in macrophage polarization.Methods A lipopolysaccharide-induced M1 macrophage polarization model was used to simulate a low-grade inflammatory response process.Quantitative real-time PCR(qRT-PCR)was employed to detect the expression levels of circHECTD1 and N6-methyladenosine(m6A)regulatory factors.Western blotting was performed to measure the expression of M1 phenotype marker proteins and autophagy-related proteins.Intracellular reactive oxygen species(ROS)levels were assessed by flow cytometry.Results Silencing circHECTD1 significantly reduced the expression of M1 phenotype-related proteins and inflammatory cytokines,decreased ROS levels,and enhanced cellular autophagy.Disordered expression of multiple m6A regulatory factors was observed in both the cellular model and peripheral blood mononuclear cells(PBMCs)from CHD patients.Significant differences in the expression levels of multiple m6A methylation regulators were identified between CHD and control groups,suggesting their potential value in CHD diagnosis.Univariate and multivariate Logistic regression analysis revealed that the m6A regulators of METTL3,METTL14,ALKBH5,and YTHDC2 were significantly associated with CHD risk.Further analysis showed that the expression of METTL14 and ALKBH5 were significantly upregulated in advanced stages of atherosclerosis.The expression level of circHECTD1 significantly correlated with FTO,YTHDF3,ALKBH5,METTL3,METTL14,and YTHDF2.In vitro experiment results demonstrated that silencing METTL14 and ALKBH5 markedly downregulated circHECTD1 expression,indicating that METTL14 and ALKBH5 positively regulated circHECTD1 expression.Conclusion Dysregulation of m6A regulatory factors may be closely associated with the occurrence and development of CHD,potentially mediated through the regulation of circHECTD1 expression and function.关键词
冠心病/circHECTD1/M1型巨噬细胞/N6-甲基腺苷Key words
coronary heart disease/circHECTD1/M1 macrophage/N6-methyladenosine分类
医药卫生引用本文复制引用
别恒杰,符亚红,贾恩志,陶正贤..circHECTD1在脂多糖诱导巨噬细胞炎症反应中的N6-甲基腺苷依赖性调控机制[J].实用心电与临床诊疗,2025,34(4):481-493,13.基金项目
国家自然科学基金资助项目(81170180) （81170180）
