清热解毒法调控PPARγ通路介导M2型巨噬细胞极化对动脉粥样硬化炎症的干预作用

Atherosclerosis (AS) is the main pathological basis of ischemic cardio-cerebrovascular disease. Inflammation runs through the whole process of AS from occurrence to development, and becomes a common link and pathway of various etiology. Intervening in inflammation is an effective and important strategy to control AS. Macrophages are the key regulatory factors of AS inflammation, M2 polarization of them is one of the important factors to inhibit AS inflammation. Activating PPARγ is an significant link to promote the M2 polarization of macrophages and is closely related to the regulation of AS inflammation. Our previous research shows that many heat-clearing and detoxification herbs have significant regulatory effects on inflammatory factor of macrophage. Here, it is hypothesized that heat-clearing and detoxification method could regulate macrophage polarization to M2 type by activating PPARγ pathway, thus inhibit AS inflammation and prevent and treat AS-related diseases. The purpose of this study was to observe the intervention effects of Huanglian jiedu decoction, a representative of heat-clearing and detoxification herbs, on the inflammation of AS by regulating the M2 polarization of macrophages in AS process，based on PPARγ pathway in vivo and in vitro. To partly reveal the role and mechanism of heat-clearing and detoxification method in regulating AS inflammation. Our study will accumulate scientific basis for improving the theory of pathogeny of heat toxin storage in AS, and preventing and treating AS and related diseases by combining with heat-clearing and detoxification method in traditional Chinese medicine.

动脉粥样硬化（AS）是缺血性心脑血管病的主要病理基础。炎症贯穿于AS发生、发展整个过程，成为多种病因的共同环节和通路，干预炎症是控制AS的有效途径和重要策略。巨噬细胞作为AS炎症反应的关键调控因素，其向M2型极化是抑制AS炎症的重要因素之一。PPARγ活化是促进巨噬细胞向M2型极化的重要环节，与AS的炎症调控密切关联。本课题组前期研究显示，许多清热解毒中药对巨噬细胞炎症因子调控作用显著。因此提出假设：中医清热解毒法可通过活化PPARγ通路调控巨噬细胞向M2型极化，从而抑制AS炎症、防治AS相关疾病。本课题拟通过体内外实验，基于PPARγ通路探讨AS过程中清热解毒代表方黄连解毒汤调控巨噬细胞向M2极化对AS炎症的干预作用，部分揭示清热解毒法调控AS炎症的作用及机制，为完善中医学关于AS热毒内蕴的病因病机理论和清热解毒法配合防治AS及相关疾病积累科学依据。

动脉粥样硬化（AS）是动脉硬化性血管病最常见的一种表现形式，是冠心病、脑血管病和血栓栓塞性疾病等缺血性心脑血管病的主要病理基础。炎症贯穿于AS发生、发展整个过程，成为多种病因的共同环节和通路，干预炎症是控制AS的有效途径和重要策略。巨噬细胞作为AS炎症反应的关键调控因素，其向M2型极化是抑制AS炎症的重要因素之一。PPARγ活化是促进巨噬细胞向M2型极化的重要环节，与AS的炎症调控密切关联。本课题组通过体内外实验，利用免疫组化、WB、RT-PCR、流式细胞术等技术方法，基于PPARγ通路探讨AS过程中清热解毒代表方黄连解毒汤调控巨噬细胞向M2极化对AS炎症的干预作用。体外研究结果显示，黄连解毒汤含药血清对RAW264.7巨噬细胞源泡沫细胞具有抑制作用，能促进胆固醇从RAW264.7巨噬细胞流出；并降低泡沫细胞炎性因子IL-1β、TNF-α水平，增加PPARγ、LXRα 、ABCG1 mRNA的表达，降低CD36 mRNA表达水平。体内研究结果显示，黄连解毒汤能显著降低动脉粥样硬化ApoE-/-小鼠血清中TC、TG、LDL-C、TNF-ɑ和CRP水平，升高HDL-C、PPARγ、TGF-β、IL-10水平，减少动脉硬化斑块。同时，黄连解毒汤干预使小鼠主动脉组织中PPARγ、Arg-1、TGF-β mRNA 表达增加，NF-κB p65 mRNA减少，使PPARγ、CD206蛋白表达升高，NF-κB p65、CD197蛋白表达明显降低。以上结果表明黄连解毒汤能通过调控PPARγ通路促使M1型巨噬细胞向M2极化，从而抑制AS炎症的发展。本研究部分揭示了清热解毒法调控AS炎症的作用及机制，为完善中医学关于AS热毒内蕴的病因病机理论和清热解毒法配合防治AS及相关疾病积累了科学依据。