miR-143抑制川崎病血管内皮损伤的作用及机制研究

Kawasaki disease (KD) is a major cause of acquired heart disease in children, and the immune response induced by T cells is a key pathophysiologic process leading to vasculitis. Our previous research suggests the increasing level of miR-143 and decreasing expression of NF-ATC1 in peripheral blood mononucletide cells （PBMC) in KD children, and bioinformatics predicts NF-ATC1 is a downstream target of.miR-143. Therefore, we proposed the hypothesis that miR143 downregulates vascular immune response by targeting NF-ATC1 in the process of vascular endothelial injury. We intend to validate this mechanism by the following research : 1) Clinical research : investigate expression levels of miR-143 and NF-ATC1 in PBMC and correlations between both of them and coronary arterial lesions and vascular endothelial injury in KD children; 2) Cell research : observe expression of miR-143 and NF-ATC1 using different stimuli to handle human umbilical vein endothelial cells (HUVEC), then evaluate effects of over expression and silencing of miR-143 on pre-handled HUVEC; 3) Analysis on miR-143 targeting NF-ATC1 gene using luciferase reporter gene systems. New mechanism of miR-143 via targeting NF-ATC1 downregulaing vascular endothelial injury in KD, which has not been reported before, is a kind of original design that can provide novel clues and potential therapeutic targets for KD children.

川崎病(KD)是儿童后天性心脏病的主要病因，T细胞活化诱导的免疫血管炎是主要病理过程。前期研究提示KD患儿血管内皮功能持续损伤；外周血单个核细胞中miR-143水平下降，NF-ATC1升高，生物信息学分析发现NF-ATC1是其下游靶标，由此我们提出理论假设：川崎病miR-143通过靶向NF-ATC1抑制血管免疫炎症。拟进行以下研究验证该假设：①临床研究：KD患儿外周血miR-143和NF-AT表达水平与川崎病冠状动脉病变和血管内皮损伤的相关性研究；②细胞研究：不同刺激物刺激人脐静脉血管内皮细胞（HUVEC）观察miR-143、NF-ATC1表达；再对刺激后HUVEC进行miR-143过表达和表达沉默处理，比较内皮细胞的损伤和NF-ATC1的表达变化；③荧光素酶报告基因系统分析miR-143对NF-ATC1基因的表达调控。该机制未见报道，具有源头创新性，可为川崎病早期预警和防控提供新的靶标。

川崎病(Kawasaki Disease)是儿童后天性心脏病的主要病因，与成年后心血管疾病密切相关。其基本病理特征是T细胞参与的全身性免疫性血管炎，钙调神经磷酸酶（Calcineurin，CaN）-活化T细胞核因子(Nuclear factor of activated T cell ，NFAT)信号通路参与的血管损害是KD发病的重要致病过程。前期研究提示KD患儿血管内皮功能功能障碍持续存在；运用系统生物学方法，明确NFAT和白细胞交互作用的调控通路在KD血管损伤中发挥作用；川崎病患儿外周血单个核细胞中NFATC1升高，本研究在此基础上，运用细胞模型，构建了miR-143过表达、表达沉默慢病毒载体，并通过荧光素酶基因报告系统，明确了miR-143对NFATC1的抑制作用；小规模研究了急性期KD患儿外周血miR-143表达水平，结果发现：KD患儿外周血miR-143明显降低，NFATC1（NFAT2）增高，为川崎病血管内皮损伤机制提供新线索，为川崎病早期诊断及早干预提供新理论依据和潜在干预靶标。