从益气活血中药调控TLR4介导炎症反应对不稳定斑块形成的影响研究益气活血配伍的抗炎机制

Acute coronary syndrome (ACS) have the clinical high fatality rate and a marked increased incidence in our country, studies have shown that toll-like receptor (TLR) 4, mediated inflammatory response directly involved in the rupture of unstable plaques, leading to the occurrence of the disease. Our group preliminary results identified yiqi huoxue compound QidanTongmai tablet (QDTM) may be affected by PI3K - Akt – eNOS signaling pathways and ICAM - l and VCAM - l gene expression to reduce the content of TNFα- , IL – 6 and inhibit the expression of TLR4, so as to exert its anti-inflammatory and myocardial protection role. Based on previous research results, we propose a hypothesis: yiqi huoxue Chinese medicine can inhibit TLR4 mediated inflammatory response and the formation of foam cells so as to inhibit the formation of unstable plaques. This study intends to establish the unstable plaque model of ApoE gene defected mice and in vitro models of foam cell formation to research the effect of QDTM on the PI3K - Akt – eNOS signaling pathways, the readjustment of TLR4 signaling pathway mediated inflammatory response and instability of the plaque through separating the prescription and discuss the possible mechanism, explore the role of yiqi and huoxue methods in this process and the interact with each other. We want to provide the experimental basis for the anti-inflammatory mechanism of yiqi huoxue Chinese traditional medicine and revealing the scientific connotation of yiqi huoxue compatibility through this study, and provide a new effective way to prevention and treatment of Sclerosis of arterial congee appearance and coronary heart disease (CHD) at the same time.

研究表明Toll样受体介导的炎症反应直接参与不稳定斑块发生、破裂，导致急性冠脉综合征（ACS）。课题组前期研究发现益气活血复方芪丹通脉片（QDTM）通过影响PI3K-Akt-eNOS信号通路以及ICAM-l和VCAM-l 基因的表达降低缺血心肌炎性因子TNFα、IL-6的含量，抑制TLR2、4表达发挥抗炎、心肌保护作用。据此，我们提出假说：芪丹通脉片能通过抑制TLR4介导的炎症反应减少泡沫细胞形成从而抑制不稳定斑块形成。本研究拟建立ApoE基因缺陷小鼠不稳定斑块模型及体外泡沫细胞形成模型，从拆方角度研究芪丹通脉片对PI3K-Akt-eNOS信号通路、TLR4信号通路介导炎症反应的调控作用及其对斑块不稳定性的影响，初步研究其分子机制，并研究益气法、活血法在此过程中的作用靶点及相互作用，为研究益气活血中药的抗炎机制及揭示益气活血配伍的科学内涵提供实验依据，同时为早期防治冠心病提供新方法。

急性冠状动脉综合征（ACS）是目前致死率最高的冠心病临床类型，研究表明Toll样受体介导的炎症反应直接参与不稳定斑块发生、破裂，导致急性冠脉综合征（ACS）。中医益气活血法是目前冠心病治疗的基本大法，课题组前期研究发现益气活血复方芪丹通脉片（QDTM）有抗炎、保护心肌作用。为证明芪丹通脉片是否通过抑制TLR4介导的炎症反应减少泡沫细胞形成从而抑制不稳定斑块形成，本研究建立ApoE基因缺陷小鼠不稳定斑块模型及体外泡沫细胞形成模型，从拆方角度研究益气中药单体、活血中药单体、益气活血单体配伍及复方芪丹通脉片对斑块稳定性、细胞泡沫化及TLR4信号通路、PI3K-Akt-eNOS信号通路的影响。. 研究结果表明：益气活血中药复方芪丹通脉片可显著下调TLR4、 NF-κB、MMP9的表达，同时下调TRL4下游炎性因子TNF-α、C反应蛋白 和 IL-6 的表达，抑制TLR4介导的炎症反应，从而减少泡沫细胞及不稳定斑块的形成。其抑制TRL4信号通路介导的炎症反应过程可能是通过激活PI3K-Akt-eNOS信号通路实现的。同时本实验通过拆方研究方法发现芪丹通脉片及益气活血中药单体配伍组对斑块的稳定作用及抑制TLR4介导的炎症反应作用优于任一单体组，其中活血组疗效优于益气组。. 本实验研究在中医经典气血理论的指导下，首次以TLR4介导炎症通路为靶点，证实了益气活血中药的抗炎作用并阐明了相关机制，为早期防治动脉粥状硬化与冠心病提供新方法。并且通过拆方研究方法丰富了中医经典益气活血配伍的科学内涵，同时为“活血法贯穿冠心病治疗始终”提供科学依据，具有非常重要的理论意义和临床价值。