血清药理学结合血清药物化学研究藏药八味沉香散抗心肌缺血再灌注损伤的作用机制及药效物质基础

Coronary heart disease is major cause of death worldwide, the treatment for coronary heart disease is timely and effective myocardial reperfusion. However,the process of reperfusion can itself induce cardiomyocyte death, known as myocardial reperfusion injury, for which there is still no effective therapy,finding an drug ,which can effectively prevent and treat of myocardial ischemia reperfusion injury is an urgent problem needs to solve.. Baweichenxiang powder is commonly used in the tibetan clinical treatment of coronary heart disease, Our previous studies found: Baweichenxiang powder preconditioning can protects myocardial ischemia and hypoxia / reoxygenation injury by inhibiting reactive oxygen species and cell apoptosis. The chemical study of single herbs of Baweichenxiang powder found that its main components are fatty acids, flavonoids, phenolic acids and terpenoids and so on.But the role and exact mechanism and effective substance of Baweichenxiang powder on myocardial ischemia-reperfusion injury have not been clarified.Therefor,we will evaluate the cardioprotective effect of Baweichenxiang powder against ischemia-reperfusion injury induced by ligation of left anterior descending coronary artery in rats, study fingerprint of serum containing Baweichenxiang powder and its protective function and mechanism in myocardial cell hypoxia/reoxygenation injury by serum pharmacochemistry and serum pharmacology method,and analyse the relationship between the fingerprint chromatogram and the cardioprotect effect to explore the effective substance of Baweichenxiang powder against ischemia-reperfusion injury. It will provide scientific experimental evidence for the prevention and treatment for ischemic heart disease using Baweichenxiang powder,develop the tibetan medicine in the field of the treatment for ischemic heart disease.

冠心病是当今世界常见死亡原因之一，再灌注是治疗冠心病的根本措施。但心肌缺血再灌注损伤（MIRI）又是再灌注治疗难以回避的问题，用药物减轻MIRI已成为研究热点。. 八味沉香散是藏医临床治疗冠心病的经典药方。前期研究发现，八味沉香散通过抑制过氧化反应和细胞凋亡保护心肌缺血和心肌细胞缺氧/复氧损伤，研究表明：其组方中各单味药的主要成分为脂肪酸类、黄酮类、酚酸类和萜类等。但八味沉香散保护心肌缺血再灌注损伤的作用和确切机制及有效物质基础尚未阐明。为此，我们拟用冠脉结扎法建立心肌缺血再灌注模型，研究八味沉香散抗心肌缺血再灌注损伤的作用；以血清药理学结合血清药物化学的方法，研究其含药血清的指纹图谱和保护心肌细胞缺氧/复氧损伤的作用及机制；通过分析血清谱－效关系，探查该药保护心肌缺血再灌注损伤的药效物质基础。为八味沉香散防治缺血性心脏病提供科学的实验依据，开拓藏药在治疗缺血性心脏病的领域。

缺血性心脏病已成为威胁人类健康的重大疾病。再灌注治疗是挽救缺血性心脏病心肌损伤的根本措施，但心肌缺血再灌注损伤（MIRI）是再灌注治疗难以回避的问题，用药物减轻MIRI已成为研究热点。藏药八味沉香散是藏医临床治疗冠心病的经典药方。但八味沉香散是否具有保护心肌缺血再灌注损伤的作用，其作用机制和有效物质基础尚未阐明。课题组采用冠脉结扎法和心肌细胞缺氧/复氧建立心肌缺血再灌注模型，以血清药理学结合血清药物化学的方法，评价了八味沉香散的抗心肌缺血再灌注损伤作用及其机制和体内药效物质基础。.结果：1.八味沉香散可降低心肌缺血再灌注损伤大鼠的血清乳酸脱氢酶 (LDH)和肌酸激酶 (CK)活性，缩小心肌梗塞范围，改善心肌病理形态及心肌细胞线粒体的损伤。.2.八味沉香散含药血清干预缺氧/复氧的H9C2心肌细胞后，明显降低细胞培养液中LDH、CK、IL-1、IL-6、IL-8、COX、TNF-α和MDA含量，增加GSH-Px、SOD、CAT和complex1含量；降低心肌细胞ROS含量，抑制心肌细胞凋亡，降低心肌细胞IL-6和TNF-α的基因表达，降低NDUFA10、Bax、Caspase-3、NF-κB、MPO和GSK-3β的基因和蛋白表达，增加bcl-2、PI3K、Akt的基因和蛋白表达。.3.从八味沉香散复方中鉴定出27个非挥发性和45个挥发性成分；给予八味沉香散后大鼠血清中共鉴定了16个非挥发性和24个挥发性成分。入血成分均能和PI3K/Akt/GSK-3β通路多个靶点自由结合。.结论：八味沉香散具有抗大鼠心肌缺血再灌注损伤作用，其作用是通过PI3K/Akt/GSK-3β信号通路调控氧化应激、炎症反应和细胞调亡而发挥的；且入血成分可能是八味沉香散的药效物质基础，并通过多组分-多靶点产生协同增效作用。此研究为其临床应用提供实验依据，对藏药临床推广应用、带动藏区经济发展有着重要意义。