动脉粥样硬化炎症过程中巨噬细胞极化与自噬行为的相互影响及黄连解毒汤的干预作用研究

Atherosclerosis (AS) is a chronic inflammatory vascular disease characterized by lipid accumulation and inflammatory cell infiltration.Inflammation occurs throughout the process of AS development,is closely related to the serious acute cardiovascular and cerebrovascular events.Regulation of inflammation has become an important strategy for the prevention and treatment of AS.Macrophages (Mφ) is a key factor of the formation process of AS and the development process of AS inflammation.The type of macrophage polarization determines its pro-inflammatory / anti-inflammatory effects on AS.The intensity of macrophage autophagy has a significant effect on the stability of AS.Traditional Chinese medicine clearing away heat and detoxicating method has broad anti-inflammatory effect.The interaction between macrophage polarization and autophagy in the process of AS inflammation and the intervention effect of heat clearing and detoxicating method of traditional Chinese Medicine need to be studied.This project will examine the changes in the main biological parameters of the AS process, which are closely related to the types of macrophage polarization, macrophage autophagy and inflammation through the experiments in vivo and in vitro,and the intervention effect of Huanglian Jiedu decoction.The aim of this study was to explore the interaction between macrophage polarization and autophagy in the process of AS inflammation, and to explore the role and mechanism of TCM clearing away heat and detoxicating method in the regulation and control of AS inflammation based on the relationship between macrophage polarization and autophagy.In order to further improve the inflammatory pathology and TCM understanding of the etiology and pathogenesis of "noxious heat theory" and "detoxification" treatment method based on AS inflammation,and to accumulate scientific evidence.

动脉粥样硬化(AS)是以脂质蓄积和炎症细胞浸润为主要特征的慢性炎性血管病变。炎症反应贯穿于AS发生、发展全过程，并与危害严重的急性心脑血管事件密切相关，调控炎症已成为防治AS的重要策略。巨噬细胞（Mφ）是AS形成及其炎症发展过程的关键因素，Mφ极化类型决定着它对AS的促炎/抗炎作用，Mφ自噬强度对AS的稳定性影响显著；中医清热解毒法具有广泛抗炎作用。AS炎症过程中Mφ极化与自噬的相互影响及中医清热解毒法的干预作用如何？本课题拟通过体内外实验，检测AS过程中与Mφ极化类型、Mφ自噬强度、炎症反应密切相关的主要生物学指标的消长变化以及黄连解毒汤的干预作用，旨在探索AS炎症过程中Mφ极化与自噬的相互影响、探索中医清热解毒法基于Mφ“极化-自噬”关联调控AS炎症的作用及部分机制，为完善AS炎症病理和中医学基于炎症反应认识AS病因病机的“热毒蕴结”理论和“清热解毒”治疗方法积累科学依据。

动脉粥样硬化(AS)是以脂质蓄积和炎症细胞浸润为主要特征的慢性炎性血管病变。炎症反应贯穿于AS发生、发展全过程，并与危害严重的急性心脑血管事件密切相关，调控炎症已成为防治AS的重要策略。巨噬细胞是AS形成及其炎症发展过程的关键因素，巨噬细胞极化类型决定着它对AS的促炎/抗炎作用，巨噬细胞自噬强度对AS的稳定性影响显著；中医清热解毒法具有广泛抗炎作用。前期研究证实黄连解毒汤能够减轻AS动物模型斑块形成、炎症反应，推测本方抑制AS与调控巨噬细胞极化-自噬-炎症反应有关，但分子机制尚不清楚。故本课题利用高脂诱导ApoE-/-小鼠AS模型，结合体外泡沫化细胞模型，采用病理切片、流式、免疫荧光/组化、Western blot、RT-PCR等技术，以巨噬细胞极化、自噬、炎症信号通路为研究重点，探讨清热解毒方药黄连解毒汤调控上述通路，改善AS斑块的作用机制，为清热解毒法辨证治疗AS提供理论和实验依据。本课题通过整体动物和细胞实验的研究发现，黄连解毒汤能够抑制AS斑块进展以及泡沫化细胞形成，能够改善AS小鼠的病理变化，调节脂质代谢，下调M1型巨噬细胞标志物iNOS、CD197的表达，上调M2型标志物Arg-1、CD206的表达，诱导自噬相关因子LC3Ⅱ表达、抑制AKT/mTOR/p70S6K信号通路，抑制促炎因子hs-CRP、TNF-α、IFN-γ、IL-1β、IL-6、IL-18表达，诱导抑炎因子IL-4、IL-10表达，调控TLR7/MyD88/NF-κB信号通路以降低炎症反应。以上结果说明黄连解毒汤可通过调控M1/M2型巨噬细胞极化、诱导自噬、降低炎症反应从而起到拮抗AS炎症的作用。这些结果为完善AS炎症病理和中医学基于炎症反应认识AS病因病机的“热毒蕴结”理论以及阐释“清热解毒法”治疗AS的科学内涵提供了客观依据。