冠心病血瘀证形成过程能量代谢机制可视化研究

It has been proved that myocardia energy metabolism has changed in the rise of blood lipid. However, previous studies of energy metabolism was limited to a certain stage in the course of the disease . The research team confirmed that BHVS of coronary heart disease was not only a dynamic process, and the use of cardiomyocyte energy substrate would present unique characteristics in the state of BHVS. Although the methodology of traditional Chinese medicine symptom research adopt the scientific concepts of complex system,the results are mostly abstract mathematical formula, lacking the interpretation of the complexity of obtaining information.Driven by system biology and based on computer,simulation technology is becoming a frontier of life science research.As a result,in this project, cardiomyocyte energy metabolism network model of BHVS of coronary heart disease in three-stage was established based on the metabolites. The key proteins that changed energy substrat (blood oxygen, fatty acid and glucose) were determined by correlation analysis. The simulation model of energy substrate and key protein was established by NetLogo platform. According to the emergency of energy substrate and key protein,the energy state of dynamic evolution of BHVS was deduced. It provided treatment thoughts about the occurrence and development of BHVS of coronary heart disease by learning the biological mechanism.

有研究表明血脂增高时心肌的能量代谢就已经发生了变化，但以往对能量代谢的研究只局限于病程中的某个阶段，本课题组证实冠心病血瘀证不仅是一个动态的过程，而且在血瘀证状态下心肌细胞能量底物利用会呈现独特的特点。对于中医证候研究以往虽采用了复杂系统科学理念，但结果呈现多是抽象的数学公式，缺乏对获取信息的“复杂性”解读，而以系统生物学为驱动，计算机为平台的仿真技术因其直观的将复杂的生物信息可视化展现，而正成为生命科学研究的前沿热点内容。为此本项目首先依据代谢产物建立冠心病血瘀证动态演变三阶段的心肌细胞能量代谢网络模型，结合关联分析，确定导致能量底物（血氧、脂肪酸、葡萄糖）变化趋势的关键蛋白。应用NetLogo平台建立能量底物与关键蛋白的仿真模型，推演冠心病血瘀证的动态变化的能量状态，通过对生物学机制的探索，为冠心病血瘀证的发生、发展提供治疗思想。

目的:通过构建冠心病血瘀证形成过程动物模型，建立能量代谢仿真模型推演冠心病血瘀证动态演变的生物学机制。.方法：①高脂饲料联合异丙肾上腺素建立血瘀阻证前期、亚血瘀证期、心血瘀阻证期三阶段动物模型，成模后分别予以养心通脉方，曲美他嗪干预。②检测每组，血脂、血流变、心电图、血糖、血氧、能量代谢指标、差异蛋白。③在F1uxExplorer平台计算心肌细胞含氧量，应用关联分析挖掘差异蛋白与氧气、能量底物关系。④将能量底物带入NetLogo平台模拟心血瘀证形成过程。.结果：①第一阶段血脂、血流变水平较正常组上调，差异具有显著性（P＜0.05）显示血瘀证前期成模；第二阶段动脉粥样硬化形成显示亚血瘀证期成模，第三阶段心电图改变显示心血瘀阻证成模。表明心血瘀阻证动态演变大鼠模型复制成功。干预后血液流变学指标、心电图均显著改善（P＜0.05），表明药物干预有效。②形成过程中心肌能量代谢紊乱，而药物干预可以有效地缓解这种状态，差异具有显著性（P＜0.05）。通过差异性检验、聚类分析得到心血瘀阻证形成过程13个差异蛋白：触珠蛋白、I型胶原蛋白α2链、凝血因子VIII、肌球蛋白-7、纤维蛋白原β、γ、α链、补体H、C3、C4A、肌酸激酶、天冬氨酸、细胞色素c氧化酶亚基7a。③关联性分析发现心血瘀阻证形成过程、中药干预氧气、血糖与触珠蛋白、纤维蛋白原有密切的关系。西药干预氧气、血糖与肌球蛋白、肌酸激酶有密切的关系。④NetLogo 显示随着心血瘀阻证发生，心肌细胞内氧气、ATP水平下降，血糖波动下调状态。药物干预后，心肌组织内氧气、ATP水平上调，中药对于能量的改善显著高于西药组。同时还模拟出心血瘀阻证形成需要90-100天。中药需29天，曲美他嗪需37天才能完全缓解心肌能量代谢障碍。.讨论：NetLogo 可直观显示三阶段心肌能量代谢障碍，各阶段机制又有不同（血瘀证前期与线粒体功能障碍相关；亚血瘀证期是因动脉粥样硬化的形成；心血瘀阻证期是补体、凝血系统的激活）。只有扶养心气，活血化瘀才能更加有效的治疗冠心病血瘀证。