miRNA-223-3p靶向线粒体介导川崎病血管炎性损伤的机制研究

Kawasaki Disease (KD) is a systemic autoimmune vasculitis, involving coronary artery and is one of the high risk factors for adult coronary heart Disease. Our previous studies have confirmed that mitochondria injury is an early event in KD endothelial inflammatory injury. We used Affymetrix microRNA chip to find out that miR-223-3p expression increased significantly in KD plasma; Overexpression of miR-223-3p could lead to endothelial cell growth limitation and mitochondrial morphological change. Bioinformatics analysis indicated that its target genes is ANXA6 which regulates mitochondrial. So we assume that miR-223-3p targets ANXA6 and regulates mitochondrial of KD vascular endothelial cell. We plan to clarify this mechanism by following researches: 1) Clinical research: investigate the relationship between miR-223-3p, ANXA6 expression level, mitochondrial function and the coronary artery injury in KD patients. 2) Cell research: study the effects of miR-223-3p overexpression and silencing on ANXA6 expression and mitochondrial morphology; the effects on mitochondria when it expression with ANXA6. 3) animal experiment: verify its regulation of ANXA6 by overexpress it to KD mice vascular endothelial cells . This mechanism hasn’t been studied yet, and may provide potential targets for early prevention and treatment of KD.

川崎病(Kawasaki Disease KD) 是全身性免疫性血管炎，累及心脏冠脉，是成年后冠心病的高危因素。我们前期实验证实：线粒体损伤是KD血管内皮细胞炎的早期事件。进一步应用芯片等技术发现：KD血浆 miR-223-3p表达明显增加；过表达miR-223-3p可导致内皮细胞生长受限及线粒体形态改变。生物信息学分析发现其靶基因是ANXA6，可调控线粒体。由此推测：miR-223-3p调控ANXA6靶向线粒体致KD血管内皮炎性损伤的机制。本项目拟进行①临床研究：miR-223-3p与ANXA6表达水平、线粒体功能改变及川崎病冠脉损伤的相关性；②体外研究：其过表达/表达沉默对内皮细胞ANXA6 和线粒体的影响；其与ANXA6共表达对线粒体影响③动物实验：通过其损伤KD血管内皮细胞，验证其对ANXA6的调控。该机制未见报道，可能为KD早期防治提供潜在靶标。

川崎病(Kawasaki Disease KD) 是全身性免疫性中小血管炎，常累及心脏冠脉，是成年后冠心病的高危因素。我们前期实验证实：线粒体损伤是KD血管内皮细胞炎的早期事件。进一步应用芯片等技术发现：KD血浆 miR-223-3p表达明显增加；过表达miR-223-3p可导致内皮细胞生长受限及线粒体形态改变。生物信息学分析miR-223-3p靶基因是ANXA6，可调控线粒体。由此推测：miR-223-3p调控ANXA6靶向线粒体致KD血管内皮炎性损伤的机制。本研究通过（1）临床实验：川崎病患儿PBMC中miR-223-3p及ANXA6基因和蛋白表达检测，KD患儿血浆中ROS水平检测，发现KD患儿血浆miR-223-3p 水平升高，ROS水平及ANXA6基因及蛋白表达均升高，且冠脉扩张组较冠脉未扩张组升高更明显，KD患儿线粒体功能有障碍。（2）体外实验：TNF-α和LPS刺激人脐静脉内皮细胞有剂量和时间依赖；TNF-α刺激内皮细胞损伤后，miR-223-3p水平升高，线粒体膜电位改变，ROS水平升高，但细胞中ANXA6水平检测较低。miRNA-223过表达后，与对照组比较无明显差异，miRNA223干扰病毒感染组与对照组比较，靶基因annexin是显著上调的。并通过荧光素酶基因报告分析系统观察到miRNA-223与ANXA6的3' UTR结合，抑制其表达。为KD早期防治提供可能的潜在靶标。