Tong Kind-Leng, Mahmood Zuhdi Ahmad Syadi, Wong Pooi-Fong
Department of Pharmacology, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia.
Department of Medicine, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia.
EXCLI J. 2024 Aug 29;23:1073-1090. doi: 10.17179/excli2024-7342. eCollection 2024.
Atherosclerotic cardiovascular diseases are the leading causes of morbidity and mortality worldwide. In our previous study, a panel of miRNA including miR-134-5p was deregulated in young acute coronary syndrome (ACS) patients. However, the roles of these ACS-associated miRNAs in endothelial dysfunction, an early event preceding atherosclerosis, remain to be investigated. In the present study, human aortic endothelial cells (HAECs) were treated with 7-ketocholesterol (7-KC) to induce endothelial dysfunction. Following treatment with 20 μg/ml 7-KC, miR-134-5p was significantly up-regulated and endothelial nitric oxide synthase (eNOS) expression was suppressed. Endothelial barrier disruption was evidenced by the deregulation of adhesion molecules including the activation of focal adhesion kinase (FAK), down-regulation of VE-cadherin, up-regulation of adhesion molecules (E-selectin and ICAM-1), increased expression of inflammatory genes (, and ) and AKT activation. Knockdown of miR-134-5p in 7-KC-treated HAECs attenuated the suppression of eNOS, the activation of AKT, the down-regulation of VE-cadherin and the up-regulation of E-selectin. In addition, the interaction between miR-134-5p and mRNA was confirmed by the enrichment of transcripts in the pull-down miRNA-mRNA complex. Knockdown of miR-134-5p increased expression whereas transfection with mimic miR-134-5p decreased FOXM1 protein expression. In summary, the involvement of an ACS-associated miRNA, miR-134-5p in endothelial dysfunction was demonstrated. Findings from this study could pave future investigations into utilizing miRNAs as a supplementary tool in ACS diagnosis or as targets for the development of therapeutics.
动脉粥样硬化性心血管疾病是全球发病和死亡的主要原因。在我们之前的研究中，一组包括miR-134-5p在内的微小RNA（miRNA）在年轻急性冠状动脉综合征（ACS）患者中表达失调。然而，这些与ACS相关的miRNA在动脉粥样硬化前期的早期事件——内皮功能障碍中的作用仍有待研究。在本研究中，用人主动脉内皮细胞（HAECs）用7-酮胆固醇（7-KC）处理以诱导内皮功能障碍。用20μg/ml 7-KC处理后，miR-134-5p显著上调，内皮型一氧化氮合酶（eNOS）表达受到抑制。内皮屏障破坏表现为包括粘着斑激酶（FAK）激活、VE-钙粘蛋白下调、粘附分子（E-选择素和ICAM-1）上调、炎症基因（、和）表达增加以及AKT激活在内的粘附分子失调。在7-KC处理的HAECs中敲低miR-134-5p可减弱eNOS的抑制、AKT的激活、VE-钙粘蛋白的下调和E-选择素的上调。此外，通过下拉的miRNA-mRNA复合物中转录本的富集证实了miR-134-5p与mRNA之间的相互作用。敲低miR-134-5p可增加表达，而转染模拟miR-134-5p可降低FOXM1蛋白表达。总之，证明了一种与ACS相关的miRNA，miR-134-5p参与内皮功能障碍。本研究结果可为未来将miRNA作为ACS诊断的辅助工具或治疗靶点的研究铺平道路。
