基于中医“瘀毒”病机的活血解毒中药对血管内皮损伤过程中ANRIL表达的影响

"Caused by blood stasis poison" leads to stasis poison each other is the key of coronary heart disease development and deterioration. On chromosome 9 p21 locus of long chain noncoding RNA - ANRIL considered independent of traditional cardiovascular risk factors and atherosclerosis (AS) closely related to the severity of the disease and prognosis, but the mechanism is still unclear. The early stage of the research found that the ANRIL used as one of target molecules of poison and blood stasis syndrome with AS, ANRIL plays an important role in the pathogenesis and prognosis of cardiovascular disease, and the pathological process of hypercoagulable state and highly inflammatory during atherosclerosis and traditional Chinese medicine "stasis poison" have many similarities. In the previous studies, the results showed that the traditional Chinese medicine can play anti-inflammatory, anti-atherosclerotic and other vascular endothelial protective effects, but it has not been reported that the tradtional Chinese medicine can regulate the expression of inflammatory cytokines through ANRIL.This research used TNF-α induced vascular endothelial cell injury and classic ApoE -/- mice carotid artery injury AS model constructing experimental system, based on the corresponding principle and used ANRIL as the breakthrough point,to explore the role of ANRIL in vascular lesions under the circumstance of atherosclerosis and the targets of TCM for activating blood detoxification, to provide experimental basis of TCM for activating blood detoxification with the effect of prevention and control atherosclerosis in the clinical application.

“因瘀致毒”导致瘀毒互结是冠心病病情发展和恶化的关键所在，研究表明位于染色体9p21位点的长链非编码RNA-ANRIL被认为独立于传统的心血管疾病危险因素，与动脉粥样硬化（AS）的严重程度及预后密切相关，但其具体机制尚未明确，课题组前期发现ANRIL是冠心病瘀毒证的目标分子之一，在急性心血管疾病的发病及预后中扮演重要角色，且AS形成过程中的高凝状态及高度炎性反应与中医“瘀毒”状态多有相似之处。课题组前期研究表明活血解毒中药具有抗炎、抗AS等血管内皮保护效应，但其是否通过ANRIL调控炎症因子表达尚未见报道。本研究借助TNF-α诱导的血管内皮细胞损伤及经典的颈动脉损伤ApoE-/-小鼠AS模型构建实验体系，基于方证对应原则，以ANRIL为研究切入点，探讨ANRIL在AS形成过程中的作用及活血解毒中药的干预效应，为活血解毒中药防治AS的临床应用提供实验依据。

尽管针对心血管疾病的相关危险因素进行药物干预及血运重建治疗可显著改善心血管疾病的预后，但冠心病仍是严重危害人类健康与生命的重大慢性疾病。其中，急性心血管事件的高度危害性仍是我们面临的巨大挑战，课题组前期提出“因瘀致毒”导致瘀毒互结是冠心病病情发展和恶化的关键所在，为急性心血管事件的防治提供了新思路。动脉粥样硬化（AS）状态下存在的高度炎性与中医“瘀毒”多有相似之处，炎症反应参与到动脉粥样硬化斑块损伤的起始和进展阶段，其中血管内皮损伤被认为是AS的起始因素和中心环节，其功能变化早于临床症状的出现并可预测心血管事件的预后。NF-kB信号通路介导的炎症过程在AS发生发展中发挥至关重要的作用，为CAD的治疗提供了潜在的治疗靶点。.本课题借助TNF-α诱导炎症性人脐静脉内皮细胞损伤构建体外实验体系，借助高脂饲料喂养的ApoE-/-小鼠构建体内AS模型，以与炎症反应显著相关的NF-kB信号通路为切入点，研究活血解毒中药活性成分配伍对炎症状态下血管内皮功能状态及相关信号通路的影响，借以阐释其潜在作用机制，明确活血解毒中药活性成分配伍对内皮细胞保护作用及AS病理过程中血管病变的干预效应。研究结果表明，活血解毒中药活性成分配伍可显著改善炎症状态下血管内皮细胞活力、降低LDH漏出率、炎症因子水平表达、下调NF-kB、YY1基因及蛋白表达，上调AMPK基因表达，但对ANRIL表达未见显著影响；体外实验与体内实验结论一致，同时发现芍药苷配伍小檗碱可降低血清血脂水平、减轻管壁厚度及AS斑块面积、减轻炎症细胞浸润等作用，说明活血解毒中药活性成分配伍可通过干预AMPK-NF-kB-YY1信号通路减轻炎症反应发挥血管内皮细胞保护及抗动脉粥样硬化作用。这种基于冠心病瘀毒病机实质研究活血解毒中药防治AS的作用机制，为活血解毒药物防治AS相关性疾病的临床应用提供科学依据。