基于mTOR信号通路介导的自噬探讨益气活血法抗动脉粥样硬化作用机制

Atherosclerosis is the major pathological foundation of many cardiovascular diseases. So far, western medicine is not able to solve the problem very well. Recent studies showed that moderate autophagy can suppress the formation and development of AS. Many recipies of benefiting qi for activating blood circulation also have the function of anti-AS effect. It’s proven by our early works that the classical recipe of benefiting qi for activating blood circulation, buyang huanwu decoction, can regulate the expression of relative autophay proteins. Based on the above researches, we proposed that the anti-atherosclerosis effect of benefiting qi for activating blood circulation is in related with autophagy, and explore the participation of the possible signaling pathways. We begin with autophagy in this study, using ApoE-/- Mouse models of atherosclerosis and in vitro cultured peritoneal macrophage to observe: ① effects of buyang huanwu decoction on autophagosome and autophagy relative proteins expression, and its effects on mTOR signaling pathway relative protein phosphorylation. ② Correlations between anti-AS effects of buyang huanwu decoction and autophagy or mTOR signaling pathway through in vivo and in vitro inhibition of autophagy. The research objectives is to identify the mechanism actions ofbuyang huanwu decoction, to illustrate the micro-mechanism of therapeutic effects of benefiting qi for activating blood circulation, and to provide research strategy of Traditional Chinese Medicine preventing and treating AS.

动脉粥样硬化( Atherosclerosis, AS)是心血管系统疾病的主要病理基础，目前的西医治疗手段仍不能很好的解决这一难题。研究表明，适度自噬能抑制AS的形成和发展；多种益气活血方剂也具有显著抗AS的作用。前期工作证实，益气活血经典代表方补阳还五汤可调节自噬相关蛋白的表达。基于此，我们提出益气活血法抗AS作用与自噬有关的假说，并探讨可能参与其中的信号通路。本课题从自噬着手，拟运用ApoE-/-小鼠动脉粥样硬化模型和体外培养小鼠腹腔巨噬细胞，观察：①补阳还五汤对自噬体的影响，对自噬相关蛋白表达的影响，对mTOR信号通路相关蛋白磷酸化的影响；②通过在体、离体水平抑制和诱导自噬，研究补阳还五汤抗AS与自噬的相关性，以及与mTOR信号通路介导的相关性。以期明确补阳还五汤抗AS的作用机理，阐释益气活血法治疗疾病的微观机制，为中医药防治AS提供新的研究思路。

动脉粥样硬化(Atherosclerosis, AS)是心血管系统疾病的主要病理基础，目前的西医治疗手段仍不能很好的解决这一难题。适度自噬能抑制AS的形成和发展；多种益气活血方剂也具有显著抗AS的作用，益气活血经典代表方补阳还五汤可调节自噬相关蛋白的表达。基于此，我们提出益气活血法抗AS作用与自噬有关的假说，并探讨可能参与其中的信号通路。. 本课题从自噬着手，运用ApoE-/-小鼠动脉粥样硬化模型、AS家兔模型和AS小鼠主动脉平滑肌细胞，观察：①补阳还五汤对AS家兔主动脉斑块面积、自噬细胞情况，血脂水平，血清中NO、eNOS水平及相关炎症因子的体的影响；②Astragaloside对动脉粥样硬化小鼠主动脉VSMCs钙化、自噬的影响，Astragaloside对VSMCs钙化、自噬的影响及与mTOR信号通路的相关性。并利用响应面法和人工神经网络法优选补阳还五汤提取工艺。. 结果表明：①补阳还五汤可通过调节斑块内细胞自噬水平，调节eNOS活化，诱导NO释放，并减少炎症因子的释放，明显降低动脉硬化斑块的斑块面积，降低TC、TG、LDL-C水平，升高HDL-C水平，达到抗动脉粥样硬化作用。②Astragaloside抑制动脉粥样硬化中血管平滑肌细胞的自噬和钙化，与过表达lnc H19和抑制DUSP5有关，ERK和mTOR信号通路参与这一过程。③BP神经网络模型法仿真优化的补阳还五汤提取工艺具有提取效率高、能耗低的优点，适合工业化大生产。. 本课题明确了补阳还五汤抗AS的作用机理，阐释以补阳还五汤为代表的益气活血法治疗疾病的微观机制，为中医药防治AS提供新的研究思路。