内质网应激通过JAZF1/AKT/mTOR信号途径调控巨噬细胞自噬影响易损斑块的作用研究

Endoplasmic reticulum stress and autophagy played an important role in the occurrence and development of unstable plaque of atherosclerosis. However, the mechanism is not clear. . Relative studies revealed that endoplasmic reticulum stress is impossible to regulate macrophage autophagy and unstable plaque of atherosclerosis via JAZF1/AKT/mTOR pathway. We found that JAZF1 overexpression improved glucose-lipid metabolism disorder and atherosclerosis. However, the molecular mechanism through which JAZF1/AKT/mTOR pathway regulates macrophage autophagy and unstable plaque of atherosclerosis is still largely unknown and warrants extensive studies. For this purpose/Therefore, we used endoplasmic reticulum stress activator to further validate the existence of JAZF1/AKT/mTOR pathway in macrophages. Moreover, using in vitro cell experiments in vivo animal experiment, the exact role of JAZF1/AKT/mTOR pathway under endoplasmic reticulum stress on regulating autophagy of macrophages was intensively analyzed.Subsequently, using in vivo animal experiment, we will explore JAZF1/AKT/mTOR pathway in autophagy and unstable plaque of atherosclerosis within endoplasmic reticulum stress. Finally, the mechanism of JAZF1 will be illuminated in macrophage autophagy through the interactions of NF-κB pathway. Nevertheless, AKT phosphorylation antibody array and AKT/mTOR signal pathway microarray were used to identify new moleculars involved in JAZF1/AKT/mTOR pathway-regulated autophagy of macrophage. This will provide new experiment and theoretical data for the mechanism of atherosclerosis.

内质网应激和自噬是影响动脉粥样硬化（AS）易损斑块的重要因素，但目前对其影响AS易损斑块的作用机制尚不明确。.前期结果提示内质网应激极可能经JAZF1/AKT/mTOR信号途径调控巨噬细胞自噬而影响AS易损斑块。课题组在国家自然基金的资助下发现JAZF1基因过表达小鼠的糖脂代谢紊乱及AS病变均得到改善。我们拟在此基础上：进一步证实内质网应激时JAZF1/AKT/mTOR 途径在巨噬细胞中的存在；细胞实验分析内质网应激通过JAZF1/AKT/mTOR途径对巨噬细胞自噬的调控作用；动物实验分析内质网应激介导JAZF1/AKT/mTOR 途径对斑块内巨噬细胞自噬及AS易损斑块的调控作用；最后解析内质网应激通过JAZF1/AKT/mTOR途径调控巨噬细胞自噬的分子机制，重点关注其影响NF-κB途径，并利用抗体芯片及基因芯片寻找更多与JAZF1/AKT/mTOR途径调控巨噬细胞自噬相关的信号分子。

动脉粥样硬化性相关疾病的发病率呈现逐年升高的趋势，特别是在老年人群中，导致其死亡的重大及常见病是急性冠脉综合征。而动脉粥样硬化易损斑块破裂是其发病的病理基础。研究发现内质网应激和自噬是影响动脉粥样硬化易损斑块的主要因素。但是，动脉粥样硬化易损斑块的发生机制目前仍未完全阐明。因此，本项目主要通过体内外实验探讨内质网应激和自噬影响动脉粥样硬化易损斑块的潜在分子机制。运用了多种实验手段，包括重组腺病毒载体、Real-Time PCR、Western Blot技术、病理切片染色、透射电镜等多种实验方法，在体外细胞试验以及动脉粥样硬化易损斑块模型小鼠中证实了如下立项假设：1）内质网应激时，中脑星形胶质细胞源性神经营养因子改变了巨噬细胞的AKT和mTOR的磷酸化水平；2）细胞实验发现内质网应激时中脑星形胶质细胞源性神经营养因子可能通过AKT/mTOR 信号通路调控巨噬细胞自噬；3) 动物实验发现内质网应激时中脑星形胶质细胞源性神经营养因子可能通过AKT/mTOR信号途径调控体内斑块巨噬细胞自噬和动脉粥样硬化易损斑块。此外，在本基金资助下我们也探讨了尼克酰胺磷酸核糖转移酶（NAMPT）对动脉粥样硬化的作用及分子机制。采用了NAMPT-RNAi的特异性腺病毒载体干预动物及细胞，研究发现：NAMPT基因表达抑制可能通过PPARα-LXRα-ABCA1/ABCG1通路促进巨噬细胞胆固醇逆向转运，从而发挥抗动脉粥样硬化作用。因此，在本基金的资助下，我们的研究为深入探讨动脉粥样硬化的分子机制及防治策略提供了新的靶点。