lncRNA AP001605募集DNA甲基转移酶影响损伤的粥样硬化动脉再内皮化的机制研究

Early re-endothelializaiton is the key for prevention of coronary stent thrombosis. In our previous study, we applicated lncRNAs microarray technology to detect and screen the specific lncRNAs of injured atherosclerotic artery, and validated them using quantitative real-time polymerase chain reaction. Results demonstrated that lncRNA AP001605 was significantly upregulated in injured atherosclerotic artery, and was closely associated with HUVECs apoptosis, down regulation of eNOS and blockage of re-endothelialization. Further RNA pull-down experiments showed that lncRNA AP001605 combined with the DNA transmethylase DNMT3b. Therefore, we speculate that lncRNA AP001605 combines with DNA transmethylase 3b directly inhibit the production of NO and EC re-endothelializaiton progress in HUVEC cells. All will finally lead to injured atherosclerotic artery damage and impaired EC re-endothelializaiton. The aim of the present study was to firstly confirm the direct combination effect of lncRNA AP001605 with the DNA transmethylase DNMT3b, and then explore the involved the possible mechanism of blockage of ECs re-endothelializaiton. This research may help to discover the new mechanism of HUVEVC apoptosis in injured atherosclerotic artery and represent potential targets for drug interventions to improve re-endothelializaiton.

促进早期血管再内皮化是防治冠脉支架内血栓的重要策略。本课题组前期研究发现，lncRNA AP001605在损伤的动脉粥样硬化血管中异常高表达，且与内皮细胞凋亡及eNOS表达异常、损伤血管再内皮化障碍密切相关。但lncRNA AP001605如何介导上述病生过程，目前尚不明确。本项目拟在前期研究基础上，采用体内实验和体外研究结合的方法，以lncRNA AP001605为切入点，从细胞、组织和整体三个层次深入研究，采用RNA-IP等手段首先证实lncRNA AP001605通过募集DNA甲基转移酶DNMT3b，使eNOS启动子甲基化从而使NO生成减少，最终影响血管再内皮化进程。本项目拟通过严谨的科学实验验证所提出的科学假设，项目成果有望对长非编码RNA AP001605影响损伤的粥样硬化动脉再内皮化的分子机制详尽阐释，为寻找损伤的粥样硬化血管再内皮化的靶点提供科学依据。