AMPK介导的细胞自噬炎性反应信号通路对痰瘀互结证AS的调控机制及化痰祛瘀解毒方药的干预作用

Atherosclerosis(AS) is one of the major diseases menacing human’s health. Autophagy is well recognized as a survival mechanism and not as a death pathway. Moreover, evidence is accumulating that autophagy occurs in advanced atherosclerotic plaques and pharmacological approaches have recently been developed to stabilize vulnerable, rupture-prone lesions and anti-inflammation through induction of autophagy. According to the pathogenesis of ‘deficient root and excessive superficial’ and the combination with ‘phlegm, blood stasis, toxicity’ in AS, meanwhile ,and based on our preliminary study that we explore the different therapeutic principle compatibility and strictly select the prescription and dose screening, we find that the Huatan Quyu Jiedu Tongmai Formula which has the function of “removing blood stasis, reducing phlegm; detoxifying, Tongmai” have a remarkable and safe effect on anti-inflammation, anti-AS . So, we propose the hypothesis that AMPK-mediated Autophagy-Inflammatory reaction Signaling Pathway might be one of the biological foundation, reducing phlegm ,removing blood stasis; detoxifying, Tongmai and the formula might be activate AMPK-autophagy-inflammatory reaction signaling pathway to anti-AS. On the animal experiment, we use ApoE-/- mouse with syndrome of intermin-gled phlegm and blood stasis AS model and on classic cell models to observe the change of AMPK signaling pathway, cell autophagy, inflammatory reaction. Therefore ,this research can provide new ideas for the Chinese Herb study in the field of autophgy, supply new methods for the clinic treatment of AS and it certainly can offer the experimental and theoretical basis to interpret and develop traditonal Chinese medicine.

动脉粥样硬化(AS)是危害人类健康的主要疾病之一。现代医学研究发现AMPK介导的自噬炎性反应信号通路在AS 中起重要作用，有望成为AS 治疗的新靶点。中医认为AS 具有本虚标实，痰、毒、瘀相互促生的病机特点，前期实验证明以‘化痰祛瘀、解毒通脉’为治法的化痰祛瘀解毒通脉方具有显著的抗炎、抗AS 作用。据此，结合相关研究进展提出科学假说“AMPK介导的细胞自噬炎性反应信号通路可能是AS痰、瘀、毒病理因素的生物学基础之一，化痰祛瘀、解毒通脉治法和配伍方药发挥的明显抗AS作用可能与激活AMPK诱导自噬---炎性反应信号传导通路密切相关。”课题拟采用ApoE-/-小鼠痰瘀互结证AS 模型及体外相关细胞病理模型，观察AMPK通路、细胞自噬、炎性反应等指标的变化，探寻复方及方中主要活性成分对自噬-炎性反应信号通路的作用环节及靶点。为认识中医药与细胞自噬及AS 的相互关系及AS的中医药防治提供实验依据。

随着社会经济的发展和国民生活方式的改变，尤其是社会老龄化及城镇化进程的加速，我国人群心血管病的危险因素普遍暴露，冠状动脉粥样硬化性心脏病的发病率持续增加。研究发现AMPK介导的自噬炎性反应信号通路在动脉粥样硬化（AS） 发病机制中起重要作用，有望成为AS 治疗的新靶点。中医认为AS 具有本虚标实，痰、毒、瘀相互促生的病机特点，与炎症关系非常的密切。本课题在前期研究的基础上，认为冠心病病机中，毒邪袭脉是痰瘀互结证病机演化的重要环节。因此创立了化痰祛瘀解毒通脉颗粒用于冠心病痰瘀互结证的治疗。通过建立ApoE-/-小鼠动脉粥样硬化痰瘀互结证动物模型，并采用化痰祛瘀解毒通脉颗粒进行干预，评价其对AS 和血脂代谢、炎症反应的影响。随后分别以J774A.1小鼠巨噬细胞和人急性单核白血病细胞为研究对象，观察化痰祛瘀解毒通脉颗粒主要有效成分对单核/巨噬细胞炎症反应和自噬的影响，并分析其作用机制。本课题结果显示，炎症反应在动脉粥样硬化发展过程中具有重要的作用。化痰祛瘀解毒通脉颗粒能够明显抑制ApoE-/-小鼠AS斑块形成及炎症反应。进一步的研究证实化痰祛瘀解毒通脉颗粒主要活性成分能够抑制单核/巨噬细胞自噬作用，从而抑制内毒素（LPS）诱导的炎症反应。其作用机制在于对LPS诱导的单核/巨噬细胞炎症反应过程中AMPK及Akt信号通路的激活作用。以上研究结果不仅进一步证实冠心病病机演变过程中痰、瘀、毒病理因素的关联性。而且从对证治疗的角度阐明了动脉粥样硬化痰、瘀、毒病机演变过程。并深入探讨中药复方中主要活性成分对AMPK诱导自噬抑制炎性反应通路抗AS作用途径，探寻化痰祛瘀解毒治法的分子机制，有助于治疗冠心病痰瘀互结证中药复方新药的研发。. 本研究参与并获得国家科技进步二等奖1项，省部级科技进步奖2项，参加国内外学术会议报告15次，获得专利授权2项，发表学术论文6篇，获得中药新药临床研究批复1项，培养硕士7名，博士3名，出站博士后4名。