【作者】 刘胤敏；

【导师】 邱明丰；

【作者基本信息】 上海交通大学 ， 中药学， 2012， 硕士

【摘要】 冠心病是由动脉粥样硬化造成冠状动脉管腔狭窄,使冠状动脉供血不足和心肌缺血,并引起心脏病变。目前已成为我国城市人口死亡原因中的第三位。西医常采用硝酸脂类、钙拮抗剂及β受体阻断剂等进行治疗,但存在头晕、面红、血压低等不良反应。因此副作用小、疗效显著的复方中药的研究越来越受人关注。复方丹参滴丸是根据中医的传统理论与现代药学新技术相结合研制成的滴丸剂,具有多组分、多环节、多靶点的心血管保护作用特点,已成为预防与治疗冠心病心绞痛的临床一线药物,但目前缺乏整体反映其作用的评价体系和标准。代谢组学的出现为这一问题的解决提供了契机。本文利用代谢组学技术研究冠脉结扎对心肌缺血大鼠内源性代谢物的影响,从整体代谢网络调控角度阐释心肌缺血对机体的影响机制,评价复方丹参滴丸干预心肌缺血大鼠的作用。结合常规药理学方法,尝试从代谢图谱角度全面认识心肌缺血的危害,阐明复方丹参滴丸干预心肌缺血的作用机制,建立以代谢组学方法评价药物疗效的方法。主要内容与结果如下:本研究采用与临床最为相似的冠脉结扎方法建立并复制―心肌缺血‖大鼠模型,通过测定体重、心脏重量及脏器比、血浆CK水平、心肌组织中MDA、SOD含量,心肌组织形态情况等,考察冠脉结扎对机体缺血损伤的影响,同时给予复方丹参滴丸进行干预,并尝试从抗自由基、抗氧化方面阐释复方丹参滴丸的作用机制,结果表明复方丹参滴丸疗效肯定,具有良好的抗氧自由基、抗氧化损伤、保护心肌的作用。在此基础上,我们采用大鼠血浆和尿液的代谢组学方法研究心肌缺血大鼠内源性代谢物的变化。结果发现通过对假手术组、模型组及模型给药不同组间,造模给药三天及二十八天不同时间点的比较,冠脉结扎导致大鼠心肌缺血与生物体的能量代谢、糖代谢、谷胱甘肽代谢、乳糖代谢、ABC转运蛋白等代谢途径都有密切关系。心肌缺血大鼠血浆中延胡索酸、琥珀酸、葡萄糖、柠檬酸、乳酸、尿素、天门冬氨酸、苏氨酸及肌酐等内源性代谢物有差异性变化,在尿液中葡萄糖、果糖、脯氨酸、苏氨酸、甘氨酸、丙二酸、尿酸、十四酸、硬脂酸等有差异性变化,而通过造模给药后,复方丹参滴丸给药的大鼠这些内源性代谢物都有不同程度的回调,说明复方丹参滴丸的有效成分能够通过改变机体代谢物的变化,改善机体代谢网络,从而发挥降低心肌耗氧、改善心肌能量代谢、保护心肌细胞能力等调节作用。更多还原

【Abstract】 Coronary heart disease(CHD) is caused by atherosclerotic coronary artery stenosis, leading to insufficient blood supply of coronary and myocardial ischemia which will result in heart attack. Currently, CHD has become the third cause of death in China. Western medicine usually treat it with nitric acid lipids, calcium antagonists and beta-blockers, but there are some side-effects, such as dizziness, flushing and low blood pressure. With little side effects and significant effect, Traditional Chinese Medicine(TCM) is getting increasing attention in current researches.Compound Danshen dripping pills(CDDP) are developed based TCM theory and modern preparation technologies. With its multi-component, multi-link and multi-target characters, CDDP got tremendous employment in the prevention and treatment of CHD in clinic. The evaluation of the whole effect of CDDP, however, still lack a overall system and standards to reflect. The appearance of metabolomics has provided an opportunity for the resolution of this issue.With the technology of metabolomics, this paper studied the change of endogenous metabolites by ischemic myocardium resulted from coronary ligation in rats. By explaining the mechanism of myocardial ischemia on the body from the perspective of the overall metabolic network, the CDDP are evaluated. Combined with conventional pharmacological approaches, we tried a comprehensive understanding of the hazards of myocardial ischemia from the perspective of metabolic pathway, clarified the mechanism of CDDP interfering myocardial ischemia, and established a metabolomics approach to evaluate drug effect. The main content and the results are as follows:In this study, we made coronary artery ligation to establish and copy the rat model of myocardial ischemia, which is similar to the clinical disease. By measuring body weight, heart weight, heart-body ratio, CK in plasma, MDA and SOD in tissue, as well as the myocardial morphology, we investigated the influences on myocardial ischemia caused by coronary artery ligation. We also treated rats with CDDP, and evaluated its effect on the mechanism of anti-oxidative and anti-free radical. The result showed that CDDP have excellent effects: anti-oxygen-radicals, anti-oxidative-damage and protection of myocardial cells. Based on this, we studies the endogenous metabolites in the plasma and urine of rats with metabolomics. By the comparison of sham group, model group and model+CDDP group, and the difference between the 3rd day and the 28th day after modeling and treating, we found that myocardial ischemia leaded by ligation is closely related to energy metabolism, glucose metabolism, glutathione metabolism, lactose metabolism and ABC transporter, etc. In the plasma of rats, 2-butenedioic acid, succinic acid, l-threonine, creatinine, l-aspartic acid, l-isoleucine, citric acid, d-glucose, urea and so on had significant changes before and after myocardial ischemia. In the urine of rats, glycine, tetradecanoic acid, d-glucose, octadecanoic acid, uric acid, l-proline, d-fructose, aminomalonic acid, l-threonine and others made differences. However, with the administration of CDDP, rat endogenous metabolites have different degrees of correction, which shows that the active ingredient of the CDDP helped to change the level of metabolites and improve the metabolism networks, which leads to decreasing myocardial oxygen consumption, regulating myocardial energy metabolism, and protecting myocardial cells.更多还原