趋炎脂质通过调控microRNA影响动脉粥样硬化形成

Our previous study found that proinflammatory lipids were related to endothelial dysfunction and atherosclerosis(AS), but the mechanisms of AS formation are still not entirely clear. Recent studies have shown that microRNAs (miRNAs) are involved in the regulation of AS formation. Our preliminary experiments showed that proinflammatory lipids can induce miRNA expression differently in the endothelial cells. Some of these different expression miRNAs may be involved in vascular endothelial cells proliferation and migration. Therefore, we hypothesized that proinflammatory lipids promote AS by regulating the miRNA expression in vascular endothelial cells. In this project, vascular endothelial cells will be treated with proinflammatory lipids. The different expression of miRNA will be found out by miRNA array、qRT-PCR、RNAi and bioinformatics. And the target genes which are related to the endothelial dysfunction and AS formation will also be searched. Finally, one of the most different expression miRNA will be chosen. Low-density lipoprotein receptor-null(LDLr-/-) mice will be treated with this miRNA and the AS formation, the target genes and proteins related to vascular endothelial dysfunction will be measured. The aim of this study is to obtain reliable evidence that proinflammatory lipids promote AS by regulating the miRNA expression and illustrate the novel mechanism of AS formation. Findings from this study may provide scientific evidence that the miRNA regulated by proinflammatory lipids are the new target for treating AS in the future.

我们前期研究发现趋炎脂质和血管内皮功能失调与动脉粥样硬化(AS) 形成有关，但具体机制尚未完全清楚。microRNA(miRNA)参与AS的调控过程。我们初步实验显示趋炎脂质能诱导血管内皮细胞的miRNA差异表达，其差异表达的miRNA可能与血管内皮细胞增殖、迁移有关。因此，我们提出趋炎脂质可能通过调控血管内皮细胞的miRNA来影响AS形成。本项目通过趋炎脂质处理血管内皮细胞，利用miRNA 芯片、qRT-PCR、RNAi、生物信息学等方法，筛选出差异表达的miRNA和寻找相应的与血管内皮功能失调和AS有关的靶基因。再挑选一个miRNA处理低密度脂蛋白受体缺乏小鼠，检测AS形成情况和相应的血管内皮功能失调的靶基因表达及蛋白活动情况。旨在获得趋炎脂质通过调控miRNA影响AS形成的可靠依据，阐明趋炎脂质促进AS形成的新机制，为将来确立趋炎脂质调控的miRNA作为新的治疗AS靶标提供科学依据。

趋炎脂质能诱导血管内皮细胞的microRNA(miRNA)差异表达，但调控机制尚未清楚。本项目通过趋炎脂质处理血管内皮细胞，利用miRNA 芯片、qRT-PCR、RNAi、生物信息学等方法，筛选出差异表达的miRNA和寻找其相应的与血管内皮功能失调有关的靶基因。在细胞、动物模型上验证趋炎脂质通过调控miRNA来促进AS形成的可能机制。项目已按原计划完成.结果发现POVPC抑制血管内皮细胞的迁移、增殖以及管状形成能力，使血管内皮源性一氧化氮合酶处于失偶联状态，与HSP90结合减少，从而使超氧自由基产生明显增加，抑制血管舒张功能。而POVPC可能通过促进miR-1278表达下调血管内皮细胞中GJA4的表达来抑制血管内皮功能，从而促进AS的形成。另外研究还发现冠心病患者中，失调的高密度脂蛋白(HDL)趋炎脂质含量增多，尤其是POVPC，并引起长链非编码的 RNAs差异性表达，而在冠心病、心脏手术患者中失调的HDL趋炎脂质可促进miR-15a-5p表达，损伤心功能。而我们分别在患者的血清标本、细胞水平和动物模型上验证，发现趋炎 HDL可以通过影响vinculin及miR-24-3P来调控血管新生。另外我们还发现先天性心脏病患者中内皮衍生颗粒中含有趋炎脂质并能引起血管内皮功能失调，而心脏瓣膜病患者，血浆中的微粒中包含趋炎脂质，在心脏体外循环手术时可诱导急性肺脏和肾脏损伤。而糖尿病患者HDL变成趋炎HDL，丧失了心肌缺血/再灌注（PTC）对心脏的保护作用。这些研究发现提示趋炎脂质的存在明显损伤血管内皮功能，促进疾病的发生、发展，影响心脏手术的预后，明确其发病机制，对确立趋炎脂质调控的miRNA作为新的治疗AS靶标，指导未来的临床研究和用药有十分重要的意义。. 在国际权威期刊JMCC、AJP- Endocrinology and Metabolism、The Journal of Thoracic and Cardiovascular Surgery、 Journal of Cardiovascular Pharmacology、Journal of Translational Medicine、BBRC、Shock、Redox Biology等发表11篇SCI文章，发表3篇中文论文，并多次在国内、外会议上交流。2篇论著在送审中。