基于AMPK/mTOR自噬轴探讨四妙勇安汤干预动脉粥样硬化机制研究

Atherosclerosis (AS) is a chronic and persistent inflammatory vascular disease, and it is the pathological basis of various cardiovascular and cerebrovascular diseases. Recent studies have shown that macrophage autophagy plays an important role in the pathophysiology of AS. Selective regulation of macrophage autophagy can promote the stability of AS plaques. AMPK/mTOR is one of the important signal pathways regulating autophagy. Previous studies and clinical data have confirmed that Si-Miao-Yong-An-Decoction has anti-inflammatory and anti-AS effects, but the mechanism has not yet been elucidated. Therefore, we hypothesized that Si-Miao-Yong-An-Decoction may play an anti-AS role by regulating AMPK/mTOR-mediated macrophage autophagy. In vitro and in vivo experiments, ApoE-/-mice AS model and ox-LDL-induced macrophage injury model were replicated to detect the AMPK/mTOR signaling pathway and downstream autophagy-specific proteins S6K1,Beclin1,and LC3. The expression of p62,and the intervention effect of Si-Miao-Yong-An-Decoction on the above mentioned parameters, aims to explore the effect and mechanism of Si-Miao-Yong-An-Decoction on the above mentioned parameters, aims to explore the effect and mechanism of Si-Miao-Yong-An-Decoction on the regulation of macrophage autophagy in the intervention of AS, and to provide an experiment for the prevention and treatment of AS by Si-Miao-Yong-An-Decoction. Based on, and accumulate scientific basis for the prevention and treatment of AS in Qingrejiedu prescriptions.

动脉粥样硬化(AS)是一种慢性持续进展的炎症性血管病变，是多种心脑血管疾病的病理基础。最新研究表明，巨噬细胞自噬在AS的病理生理中扮演着重要角色，通过选择性的调控巨噬细胞自噬能够促进AS斑块的稳定性。AMPK/mTOR信号转导通路的激活在自噬中发挥关键的调节作用。前期研究和临床资料均证实了四妙勇安汤具有抗AS等作用，但机制尚未阐明。因此，我们假说四妙勇安汤可能通过调节AMPK/mTOR介导的巨噬细胞自噬而起到抗AS的作用。在体和离体实验相结合，拟复制ApoE-/-小鼠AS模型和ox-LDL致泡沫细胞模型，检测AMPK/mTOR信号通路及其下游自噬特异性蛋白S6K1、Beclin1、LC3、p62的表达，并检测四妙勇安汤对上述参数的干预效应，旨在探索四妙勇安汤基于调控巨噬细胞自噬干预AS的作用与机制，为四妙勇安汤防治AS提供实验依据，并为清热解毒类方药防治AS积累科学依据。

动脉粥样硬化(AS)是一种慢性持续进展的炎症性血管病变，是多种心脑血管疾病的病理基础。巨噬细胞自噬在AS的病理生理中扮演着重要角色，通过选择性的调控巨噬细胞自噬能够促进AS斑块的稳定性。前期研究和临床资料均证实了四妙勇安汤具有抗AS等作用，但机制尚未阐明。本课题采用了在体动物实验和离体细胞实验相结合，复制了ApoE-/-小鼠AS模型及ox-LDL致泡沫细胞损伤模型，采用了透射电镜、病理切片、免疫荧光/组化、RT-PCR、Western blot等技术，检测了四妙勇安汤对AS小鼠及巨噬细胞自噬以及炎症反应的影响，旨在探索四妙勇安汤基于调控巨噬细胞自噬干预AS的作用与机制，为四妙勇安汤防治AS提供实验依据。本课题研究发现，四妙勇安汤能够抑制AS斑块以及泡沫细胞形成，能够改善AS斑块病理变化，调节脂质代谢，诱导自噬小体产生，调控了AMPK/mTOR/p70S6K及AMPK/ULK1自噬信号通路，增强了自噬相关因子LC3Ⅱ、Atg5、Beclin1的表达、抑制了p62的表达，降低了促炎因子IL-6、IL-18、IFN-γ和CRP的过表达，诱导了抑炎因子IL-10的表达。以上结果说明四妙勇安汤可通过调控巨噬细胞自噬、降低炎症反应从而起到改善AS的作用。这些结果为四妙勇安汤防治AS提供实验依据，并为清热解毒类方药防治AS积累科学依据。