基于“心受气于脾”探讨脾虚痰浊巴马猪microRNA155-SOCS1轴介导冠脉内皮免疫炎症的调控机制

Our research group was guided by the theory of the heart receiving qi from the spleen in Inner Canon of Huangdi.We were also inspired by the theory of preventive treatment of diseases in TCM.We moved the focus of the study of atherosclerosis formerly,and found that the Chinese medicine of invigorating the spleen and eliminating phlegm had the functions of regulateing microRNA155 level, improving the function of coronary endothelial and inflammation level, delaying the formation of atherosclerotic plaque in preliminary study of our group.But the mechanism is still unclear. Now we put forward the following hypotheses based on previous research and combination with the research progress at home and abroad: dysfunction of spleen in transport can cause endogeny of phlegm turbidity,which can lead to blood stasis. It can touch off and aggravate the immune inflammatory response of coronary atherosclerosis,then make injury to vessel endothelial cell. Thus the pathogenic factor of phlegm-turbid can enter inner membrance.It can lead to abnormal expression of endothelial microRNA155 and target gene SOCS1,then activate signal channel of JAK/STAT.This can cause differential disorders of Th cell subsets and secretion of proinflammatory cytokines,which accelerate the development of AS plaque, obstruct qi and blood circulation in vessel system.Finally it can lead to blockage stasis of heart vessel.Chinese medicine of invigorating the spleen and eliminating phlegm can affect the pathological mechanism above,and play a role in the prevention and treatment of atherosclerosis. Our project will replicate the atherosclerosis model of minipigs.Clarify the scientific connotation of the heart receiving qi from the spleen in Inner Canon of Huangdi from organ, tissue, cell and molecular level. Expatiate the relationship between dysfunction of spleen in transport and blockage stasis of heart vessel in TCM.Provide scientific basis for treating coronary heart disease based on spleen in TCM,and enrich viscera theory of spleen.

以《内经》“心受气于脾”理论为指导，以中医药“治未病”理念为启示，将动脉粥样硬化研究重心前移。课题组前期研究发现健脾祛痰中药能够调节microRNA155水平改善冠脉内皮功能及炎症水平，延缓粥样斑块形成，但作用机制仍不明确。基于前期基础结合国内外研究进展，提出假说：脾失健运，痰浊内生，因痰致瘀，触发和加剧冠状动脉粥样硬化免疫炎症反应，引起血管内皮损伤，痰浊之邪进入内膜，致内皮下microRNA155及靶基因SOCS1表达异常，活化JAK/STAT信号通路，致Th细胞亚群分化失衡，分泌促炎性细胞因子而加速As斑块发生发展，阻碍脉道气血运行，心脉阻滞。健脾祛痰中药可能影响上述病理机制防治动脉粥样硬化。本项目将复制巴马猪动脉粥样硬化模型，从器官、组织、细胞、分子水平阐明《内经》“心受气于脾”的科学内涵，阐述“脾虚生痰”与“心脉阻滞”的关系，为中医“从脾论治”冠心病提供科学依据，丰富脾脏象理论。

1.项目背景：以《内经》“心受气于脾”理论为指导，以中医药“治未病”理念为启示，结合课题组前期研究发现健脾祛痰中药能够调节microRNA155水平改善冠脉内皮功能及炎症水平，本项目提出假说：脾失健运，痰浊内生，触发冠状动脉粥样硬化免疫炎症反应，致内皮下microRNA155及靶基因SOCS1表达异常，活化JAK/STAT信号通路，分泌促炎性细胞因子而加速As斑块发生发展。.2.主要研究内容：体内实验：建立动脉粥样硬化巴马猪/大鼠模型，通过健脾化痰法干预，探讨对血清炎症因子、冠脉血管内皮、心肌结构及功能和冠状动脉粥样硬化斑块的影响机制以及保护血管内皮细胞、延缓动脉粥样硬化炎症进展的效应机制。体外实验：通过密度梯度法、Western-blot、RT-PCR等方法阐述脾失健运、痰浊内生对冠状动脉粥样硬化斑块的影响机制。.3.重要结果：健脾祛痰中药从组织、细胞、分子、细胞水平阐明其可能是通过microRNA155-1 SOCS1轴来保护冠脉血管内皮、延缓动脉粥样硬化炎症进展的。.4.关键数据：模型组血脂和炎症因子水平升高，PBMCs细胞炎症因子水平升高，血管超微结构发生改变，脂质形成，炎性细胞浸润，主动脉SOCS1、p-JAK1和p-STAT1蛋白表达降低，组织miR155表达升高、SOCS1 mRNA表达下降，PBMCs细胞SOCS1、p-JAK1、p-STAT1蛋白表达降低、miR155和的表达升高、SOCS1 mRNA表达下降；中药组和西药组血脂和炎症因子水平降低，PBMCs细胞炎症因子水平降低，脂质减少，主动脉SOCS1蛋白表达升高，组织的miR155的表达下降，SOCS1 mRNA表达上升，PBMCs细胞中p-JAK1和p-STAT1蛋白表达升高，PBMCs细胞中miR155表达下降；西药组可改变炎性细胞浸润，PBMCs 细胞的SOCS1 蛋白表达升高；中药组PBMCs细胞中SOCS1 mRNA 表达上升。.5.科学意义：本项目将动脉粥样硬化研究重心前移，复制巴马猪动脉粥样硬化模型，探讨脾失健运，痰浊内生对动脉粥样硬化早期病理变化的影响机制，从器官、组织、细胞、分子水平阐明《内经》“心受气于脾”的科学内涵，阐述“脾虚生痰”与“心脉阻滞”的关系，为中医“从脾论治”冠心病提供科学依据，丰富脾脏象理论，促进中医学术水平进步