从表观遗传学角度探讨动脉粥样硬化的中医瘀毒病机及活血解毒中药的干预

The pathogenesis of atherosclerosis (As) is a hotspot of cardiovascular disease studies in TCM. In TCM theory, it is the main pathogenesis of the incidence of plaque instability that blood stasis binds to toxican, and toxican induces acute clinical events. However, it is unclear that what phase and what mechanism blood stasis binding to toxican induces the incidence of diseases. Recent studies show that abnormal DNA methylation is an important mechanism for the incidence of As. Our previous studies showed that the representative prescriptions with the effect of activating blood circulation, and detoxicating and their compatibility may exert the effect of stabilizing plaque by increasing DNA methylation and methylase level of ApoE-/- mouse in serum. Therefore, we proposed a new hypothesis: " In the transition process from stable plaque to unstable plaque, the level of DNA methylation and aberrant methylation of related genes plays an important role, the imbalance of DNA methylation level is the important mechanism to explain blood stasis binding to toxican inducing disease at different stages of As. "Therefore, this study is designed to adopt ApoE-/ - mice as As experimental model, copy the animal models of stable plaque and unstable plaque through high-fat feeding at different periods. DNA methylation chips combined with RNA microarray are used to investigate the differences of DNA methylation and gene expression in aortic artery of ApoE-/- mice, and furtherly, the intervention effect of the representative prescriptions with the effect of activating blood circulation, and detoxicating (Xuefuzhuyu particles, Sijisanhuang capsules) and their compatibility will be observed. It will provide a scientific basis to test TCM pathogenic hypotheses of As and the prevent As by using the prescriptions with the effect of activating blood circulation and detoxicating.

动脉粥样硬化（As）的病机是中医心血管病研究的热点。中医理论认为瘀毒互结，因毒致变是导致As斑块不稳定的主要病机。然而，瘀毒在As病程通过哪些环节致病尚不清楚。新近发现，DNA甲基化异常是As发病的重要机制。我们前期发现活血、解毒方药配伍可能通过提高ApoE-/-小鼠血清DNA甲基化和甲基化酶起稳定斑块的作用。 因此提出新假说："As 斑块从稳定到不稳定状态的转变过程中，DNA甲基化水平及相关基因的异常甲基化起重要作用。DNA甲基化失衡是阐释瘀毒在As不同阶段致病的重要机制。"因此，本课题以ApoE-/-小鼠为观察对象，通过不同时期高脂喂养，复制稳定斑块和不稳定斑块的动物模型，采用DNA甲基化芯片联合基因表达谱芯片研究其DNA甲基化机制的差别，并在此基础上，观察活血、解毒代表方（血府逐瘀颗粒、四季三黄胶囊）及其配伍的干预作用，这将为验证中医瘀毒致病假说和活血解毒方药防治As提供科学依据。

动脉粥样硬化（As）的病机是中医心血管病研究的热点。活血解毒是中医干预As的重要治法。新近发现，DNA 甲基化异常是As 发病的重要机制。我们前期发现活血、解毒方药配伍可能通过提高ApoE-/-小鼠血清DNA 甲基化和甲基化酶起稳定斑块的作用。 因此提出新假说：“As 斑块从稳定到不稳定状态的转变过程中，DNA 甲基化水平及相关基因的异常甲基化起重要作用。DNA 甲基化失衡是阐释瘀毒在As 不同阶段致病的重要机制。” 因此，本课题以ApoE-/-小鼠为观察对象，通过不同时期高脂喂养，复制稳定斑块和不稳定斑块的动物模型， 采用DNA 甲基化芯片联合基因表达谱芯片研究其DNA 甲基化机制的差别，并在此基础上，观察活血、解毒代表方（血府逐瘀颗粒、四季三黄胶囊）及其配伍的干预作用。结果显示：经基因芯片和DNA甲基化芯片联合筛选，发现As 斑块从稳定到不稳定状态的转变过程中， IRS-1是重要的调控基因，PI3K/Akt是重要的信号调控通路，活血解毒中药联合应用，可明显减低As小鼠血脂，主动脉As斑块，和整体甲基化水平，并可能通过抑制PI3K/Akt信号通路，抑制IRS-1表达起到抗As作用。在课题执行过程中，发表SCI论文4篇，国内核心期刊论文4篇。申报专利2项，其中1项已授权。并以此课题为基础延伸，成功申请到国家自然科学基金面上项目等课题4项。