不同处理方式下艾燃烧生成物调节Rho/Rock激酶信号通路干预动脉粥样硬化的机制研究

Atherosclerosis is the main pathological basis of cardiovascular and cerebrovascular diseases，which are major chronic diseases threatening human health. A large number of clinical and basic studies have shown that moxibustion played a certain role of regulating lipid metabolism on the improvement of atherosclerosis.Our previous studies found that moxibustion and moxa smoke（the major component of moxa combustion products） could delay the pathogenesis progress of atherosclerosis by regulating cholesterol reverse transport.There may be certain active ingredients in moxa combustion products(MCP) and these active ingredients are important factors of moxibustion therapy against atherosclerosis.Deepening the research to active ingredients in MCP is an important breakthrough point to reveal moxibustion mechanism and optimize traditional moxibustion therapy.The combustion products are mainly composed of volatile oil,light components (gaseous substances) and heavier components (liquid and particulate matter) produced in the combustion process.In our previous study，conventional moxa smoke were used as interventions，but there is a certain distance from the discovery of the active ingredients.Rho/ROCK signaling pathway can promote atherosclerosis by inhibiting cholesterol reversal,which is the key target for our further research.This research intend to separate four forms of MCP so as to observe the effect of moxa smoke under different conditions on atherosclerosis as well as adjusting the Rho/ROCK signaling pathways in ApoE-/- mice.Further more，to explore the existence form and range of the active ingredients in moxa combustion products.

心脑血管疾病是威胁人类健康的重大慢性疑难疾病，动脉粥样硬化（AS）是其最主要的病理基础。大量临床及基础研究表明艾灸具有确切的调节血脂改善AS的作用。课题组前期研究发现，艾灸及艾烟（艾燃烧生成物的主要成分）可通过调节胆固醇逆转运改善AS。艾燃烧生成物（MCP）中存在一定的活性物质，是艾灸抗AS的重要起效因素。深入研究MCP中活性成分是揭示艾灸作用机理及优化传统灸疗的重要切入点。MCP由艾叶挥发油、燃烧过程中产生的轻组分（气态物质）及重组分（液态、颗粒物质）组成，课题组前期研究以常规艾烟为干预手段，这与揭示其中的活性物质还有一定的距离。Rho/ROCK信号通路介导了胆固醇逆转运途径，是AS发生的关键环节，也是深化前期研究的重要靶点。本课题拟采取不同方式处理分离形成四种形式MCP，观察其对APOE-/-小鼠AS病变及Rho/ROCK信号通路影响，初步界定MCP中改善AS活性物质的存在形式和范围。

本项目观察不同处理方式下艾燃烧生成物对APOE-/-小鼠AS病变及ROCK-LXRa-ABCA1信号通路的影响，探究艾燃烧生成物改善AS的机制和起效成分。本项目分别采用普通艾烟、滤过艾烟、艾绒挥发物和艾叶精油干预APOE-/-小鼠，比较干预后各组小鼠血脂水平、主动脉病变、主动脉ROCK-LXRa-ABCA1通路mRNA及蛋白表达的变化，并使用GC-MS技术、热裂解技术检测不同处理方式下艾燃烧生成物的成分差异。研究结果显示：1.普通艾烟改善AS小鼠主动脉病理结构，降低血浆TC、TG、LDL-C、ox-LDL含量，升高HDL-C含量，疗效高于滤过艾烟、艾绒挥发物、艾叶精油。2.普通艾烟能够抑制Rho/ROCK通路表达，上调PPARγ-LXRα-ABCA1通路表达，而该效应在注射Rho激动剂后减弱。3.艾烟与常规药物能在不同程度上调节ApoE-/-小鼠血脂水平和主动脉病理结构，调节主动脉ROCK-LXRa-ABCA1通路mRNA及蛋白表达。4.不同处理方式下艾燃烧生成物的成分差异研究表明，艾叶精油与艾绒挥发物的成分相似，包括桉油精、α-侧柏酮、n-棕榈酸、十氢二甲基甲乙烯基萘酚、冰片、石竹素、樟脑、(-)-4-萜品醇、α-松油醇等。与低温艾绒挥发物比较，滤过艾烟的组分还含有艾绒燃烧后生成的新物质，如乙酸、苯酚、戊醛、乙酸甲酯、2-丁酮、乙醛、新植二烯、反式-2-壬烯醛、2-呋喃甲醇、对苯二酚、γ-巴豆酰内酯、异氰基丁烷等。综上所述，不同处理方式的艾燃烧生成物能在不同程度上降低ApoE-/-小鼠血脂水平，改善主动脉病理结构，普通艾烟和滤过艾烟的疗效高于艾绒挥发物和艾叶精油；一定处理方式下的艾燃烧生成物可能通过抑制Rho/ROCK通路，激活PPARγ-LXRα-ABCA1通路，促进胆固醇逆转运，改善动脉粥样硬化；普通艾烟与常规药物均能改善AS病变，且侧重点不同；艾燃烧生成物中改善AS的活性物质主要为艾绒燃烧后生成的物质，提示艾绒的燃烧环节可能是艾灸发挥作用的关键步骤。