miR-146a/b靶向调控TLR4/NF-κB/CXCL16信号通路在LAA型脑梗死的作用机制研究

It is widely believed that atherosclerosis is an inflammatory disease with immune responses during both its initiation and progression, and atherosclerosis is the most important and main mechanism of ischemic stroke. Toll-like receptor 4 plays important roles in the development and progression of atherosclerotic lesion, and TLR4/NF-κB/CXCL16 signal transduction pathway was presumed as the tie between infection and atherosclerosis. .MicroRNAs (miRNAs) are an emerging class of highly conserved, non-coding small RNAs that regulate gene expression on the post-transcriptional level by inhibiting the translation of protein from mRNA or by promoting the degradation of mRNA. Recent works have proposed that miRNAs play crucial roles in the pathological process of atherosclerosis. Also, our detection of serum miRNAs with high-throughput genome sequencing technology showed aberrant expression of miR-146a and miR-146b-5p, which may be related with TLR4/NF-κB/CXCL16 signal transduction pathway by target gene prediction or literature search on pubmed..To explore the correlation between miR-146a/b and TLR4/NF-κB/CXCL16 signal transduction pathway, and their roles in stroke of large artery atherosclerosis subtype（LAA）, we are going to detect the expression of TLR4、NF-κB、TRAF6、IRAKl、CXCL16 mRNA and protein in peripheral blood mononuclear cells from the patients with LAA using methods of real-time quantitative reverse transcription polymerase chain reaction（qRT-PCR）and West-blot respectively. Furthermore，we explore the regulation targets of miR-146a/b on TLR4/NF-κB/CXCL16 signal transduction pathway in human umbilical vein endothelial cells induced by lipopolysaccharide, and test their roles invivo. This would be helpful to elucidate the regulation mechanism of miRNAs on TLR4/NF-κB/CXCL16 signal transduction pathway in atherosclerosis, which may be helpful to find new targets for the prevention and therapy of LAA.

目前认为动脉粥样硬化（AS）是炎症性疾病，是缺血性脑卒中的主要发病机制。我们以往的研究显示CXCL16等多种炎症因子与大动脉粥样硬化型脑卒中（LAA）密切相关,提示炎症机制在LAA发病过程中发挥重要作用。本课题着眼于TLR4/NF-κB/CXCL16信号通路在AS炎性过程中的作用，结合前期高通量基因测序结果：LAA患者血浆miR-146a/b表达显著下降，进行miR-146a/b与该通路中炎症因子和脑动脉粥样硬化程度相关性的临床研究；应用LPS诱导HUVEC体外实验明确miR-146a/b对该途径的调控靶点及作用机制；并通过在体实验证实其对颈动脉AS的干预作用，明确miR-146通过TLR4/NF-κB/CXCL16信号通路在血管内皮炎症过程中的负反馈调控作用，为阐明脑动脉粥样硬化的分子发病机制夯实理论基础，为LAA的防治寻找新的靶点。

目前认为动脉粥样硬化（AS） 是炎症性疾病， 是缺血性脑卒中的主要发病机制， 而miR-146a/b在炎症反应过程中发挥重要调控作用。 本项目着眼于TLR4/NF-κB/CXCL16信号通路在AS炎性过程中的作用，结合前期高通量基因测序结果：LAA患者血浆miR-146a/b表达显著下降，进行miR-146a/b与该通路中炎症因子和脑动脉粥样硬化程度相关性的临床研究；应用LPS诱导HUVEC体外实验明确miR-146a/b对该途径的调控靶点及作用机制；并建立颈动脉粥样硬化模型， 在体证实miR-146a/b对颈动脉AS的干预作用，阐明了脑动脉粥样硬化的分子发病机制，为LAA的防治找到了新的靶点。