基于生理药动学模型探讨丹参川芎对药保护MIRI的药效物质及配伍机制

Myocardial ischemical reperfusion injury (MIRI), one of subsequent symptom after a cardiovascular surgical operation, is inextricably involved. Traditional Chinese medicines is short of scientific expatiation about the microanalysis and effective substances, which become the rapid development of the bottleneck of traditional Chinese medicines. The results of the previous research by the applicant showed that Salvianolic acids such as Salvianolic acid B in Salviae Miltiorrhizae Radix et Rhizoma can protect MIRI and the effective components of Chuanxiong Rhizoma promote the before-mentioned cadioprotection of MIRI. Therefore, the applicant put forth a academic hypothesis about scientific expatiation of compatibility of Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma drug partnership, the academic hypothesis consider Salviae Miltiorrhizae Radix et Rhizoma as sovereign drug and the other Chuanxiong Rhizoma Chuanxiong as assistant and guide drug. The objective of this research is to elucidate the effective substances and compatibility mechanism of Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma drug partnership to protect MIRI. The research select Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma drug partnership as investigation object. The quantitative compatibility of Salvianolic acid B and the effective components of Chuanxiong Rhizoma is based on the on myocardium injury induced by ischemia-reperfusion in isolated rat heart to investigate the effective substances of Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma drug partnership. Ultra-performance liquid chromatography (UPLC)-MS/MS techinique is fast, sensitive and accurate to be proposed for the determination of effective substances such as Salvianolic acid B and Ferulic acid. The rat PBPK modeling is to establish for the whole pharmacokinetics of Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma drug partnership. According to pharmacodynamics and the quantity changes of these compounds, scientific expatiation of compatibility of Salviae Miltiorrhizae Radix et Rhizoma-Chuanxiong Rhizoma drug partnership were illustrated and the compatibility reasonability was discussed. This study will enrich the modern pharmaco-therapeutics, push forward the scientific explanation of compatibility of medicines in a prescription, create new ideas for the development of new Chinese medicines and build up a theoretical basis for the innovation of Chinese medicines.

心肌缺血-再灌注损伤(MIRI)是心脏术后难以解决的并发症,丹参川芎对药在动物药理实验和临床疗效研究方面均具有较好的保护MIRI药理作用，但缺乏对其药效物质和配伍机理的科学阐释。申请者的前期研究结果表明：丹参中有效成分丹酚酸B能有效保护MIRI，且川芎有效组分能显著促进丹酚酸B保护MIRI的作用，因此提出丹参川芎对药配伍的科学内涵-"丹参治心、川芎引经"的学术假说。本研究在前期研究的基础上，以丹参川芎对药为研究对象，采用大鼠离体MIRI模型，优化丹酚酸B与川芎中阿魏酸等有效组分的量化配伍，研究其配伍的药效物质；建立大鼠8个生理组织器官和2个血液室的生理药动学(PBPK)模型，采用现代色谱-质谱联用(UPLC-MRM）的快速分析技术，系统研究丹酚酸B配伍川芎有效组分前后的整体药代动力学变化情况，最后整体药代动力学结合药效学验证丹参川芎对药配伍的科学内涵，为推演在人体的药代动力学奠定实验基础。

本课题组基于PBPK模型，通过对丹参川芎对药配伍前后4种药效成分在血浆及心、肝、脾、肺、肾、肌肉、脑等组织中的分布差异，基于药动学参数探讨丹参川芎配伍机制-“丹参为君、川芎引经报使”的学术假说。在溶剂优化提取和述职纯化获得药效组分丹参多酚酸和阿魏酸类组分的基础上，进行了保护心肌缺血再灌注损伤的活性研究；同时基于HPLC测定阿魏酸和丹酚酸B的基础上，探讨了丹参川芎配比，阿魏酸与丹参多酚酸联合用药对药效成分阿魏酸及丹酚酸B药动学的影响；基于PBPK模型，建立了血浆及7种组织中4种酚酸类化合物（丹酚酸B、紫草酸、阿魏酸和迷迭香酸）UPLC-MS/MS的检测方法，结果显示配伍给药，大鼠血浆中4个多酚酸类药效成分的药代动力学参数均存在显著性或极显著性差异，例如4个药效成分的AUC(0-t)均不同程度增加，其中阿魏酸、丹酚酸B、紫草酸均极显著性增加（p < 0.01），配伍给药阿魏酸的AUC(0-t)是川芎组的2倍，丹酚酸B AUC(0-t)比丹参多酚酸组提高了40.50%，紫草酸比丹参多酚酸组提高了16.83%，说明两药配伍比单独用药提高了相应药效成分的生物利用度；4种酚酸类成分在大鼠组织器官的分布情况差异显著，但主要分布的器官分别：排泄器官肾、代谢器官肝脏、血液转输站肺脏、作用器官心脏，这些组织器官中4种酚酸类成分的浓度均较高。丹参-川芎配伍对4种酚酸类成分在各组织器官的影响：明显增加在心、肺、肾脏的药物浓度和生物利用度，其中以阿魏酸和丹酚酸B的影响最为显著，提高幅度在7%至173%，其中配伍后以对心脏影响最大，阿魏酸的提高了173%、丹酚酸B提高95%，迷迭香酸94%之多。丹参川芎两药配伍能提高保护心肌的疗效，而这种疗效的提高体现在两药能相互促进相应药效成分在体内吸收、分布、代谢、排泄等诸多方面，降低清除率、提高药物浓度、延长体内作用时间，极大提高药效成分的生物利用度。可见川芎对君药丹参的引经作用之大，同时也表明丹参多酚酸类成分也能提高阿魏酸在心脏的浓度和生物利用度。本项目提出提出的“丹参为君、川芎引经报使”的学术假说是合理的。