基于PK-PD-代谢组学实时动态偶联的淫羊藿-川芎药对治疗骨关节炎的配伍机制研究

Osteoarthritis (OA) is a common degenerative disease, so far, there is no effective drugs to repair cartilage and prevent the disease from developing. The preliminary study of project shows that: The compatibility of epimedium and rhizome ligustici wallichii can make a reversion to OA phenotype, lower the expression of MMP-13， and regulate the metabolic profiles of OA to normal reversal. Therefore, we come up with a hypothesis that the compatibility of Epimedium and rhizome ligustici wallichii can promote the group of medicinal compositions release, inhibite the inflammatory cytokines secretion, downregulate the expression of MMPs, and then regulate the variation of endogenous component of OA, achieve to repair cartilage and give a radical treatment to OA. The project adopts the OA model of ligament and meniscus injury, then intervened by the compatibility of Epimedium and rhizome ligustici wallichii, epimedium, rhizome ligustici wallichii respectively, using LC-MS, GC-MS to real-time detect the dynamic changes in vivo of endogenous component and medicinal composition group after molding and dosing: Simultaneous proceeding the dynamic efficacy study of MMPs, inflammatory factors and so on. Fitting the model of PK-PD and BM-PD with genetic algorithm and artificial neural network, clear the correlation between efficacy and the dynamic changes in vivo of endogenous components and medicinal composition group of OA for compatibility, PK-PD-metabonomics real-time dynamic/synchronous correlation reveals the mechanism and the material basis of treating OA by using double-drugs compatibility of tonifying kidney and activating blood epimedium and rhizome ligustici wallichii, interpreting the scientific connotation of “Bushen Huoxue” treating principle, in order to provide a basis of new drug research and development for repairing cartilage essentially.

骨关节炎(OA)是常见的关节软骨退行性疾病，至今尚无修复软骨、阻止病程发展的有效药物。项目组前期研究显示：淫羊藿-川芎配伍可逆转OA表型；下调MMP-13表达；调控OA代谢轮廓向正常逆转。因此提出假说：淫羊藿-川芎配伍促进药效成分群释放，抑制炎性因子分泌，下调MMPs表达，进而调控OA内源性成分的变化，修复软骨，治疗OA。本项目采用韧带-半月板损伤OA模型，以淫羊藿-川芎及淫羊藿、川芎干预，LC-MS、GC-MS实时检测造模及给药后内源性成分和药效成分群的体内动态变化；同步进行MMPs、炎性因子等动态药效研究，以遗传算法加人工神经网络拟合PK-PD、BM-PD模型，明确配伍对OA内源性成分、药效成分群体内动态变化与药效的相关性，PK-PD-代谢组学实时动态/同步关联揭示补肾活血药对淫羊藿-川芎治疗OA的配伍机制和物质基础，诠释“补肾活血”治则的科学内涵，为从本质修复软骨的新药研发提供依据。

本项目采用交叉韧带-半月板损伤建立骨关节炎（OA）大鼠模型，分别给予补肾活血药对淫羊藿-川芎及淫羊藿、川芎单药，LC- MS实时检测造模及给药后血浆中内源性成分和淫羊藿苷、宝藿苷I、朝藿定A/B/C、藁本内酯、洋川芎内酯A等药效成分的动态变化；同步进行MMPs、炎性因子等动态药效研究，拟合PK-PD模型，探究淫羊藿、川芎配伍对OA内源性成分、药效成分群体内动态变化与药效的相关性。给药各组代谢轮廓变化及对标志物的定量回调均表明淫羊藿-川芎配伍对OA大鼠代谢的调控作用优于淫羊藿、川芎，与组织病理学、MMP-13评价的药效一致；初步获得12个OA生物标志物，可能通过调控参与甘油磷脂代谢、α-亚麻酸代谢、花生四烯酸代谢的12个代谢产物起到治疗OA的作用，从代谢组学层面证明淫羊藿、川芎配伍治疗OA的科学合理性，初步揭示淫羊藿-川芎对OA大鼠的作用机制。淫羊藿、川芎配伍与单药各成分的药-时曲线和药动学参数存在明显差异，淫羊藿-川芎配伍后主要成分在OA大鼠体内的吸收增加、达峰时间缩短、达峰浓度增加、滞留时间延长、清除率减小、代谢减缓，从物质基础层面阐明了淫羊藿-川芎配伍治疗OA的协同作用；同步药效显示：淫羊藿-川芎药对配伍可使大鼠血浆MMPs、炎性因子等指标显著下降，药效强度显著增强，且维持较长药效时间，从药效评价角度明确了淫羊藿-川芎配伍治疗OA的增效作用。淫羊藿-川芎药对与单药在OA大鼠体内的PK-PD拟合模型均符合二房室药动模型、Sigmoid Emax拟合模型，淫羊藿-川芎配伍后Emax较单用淫羊藿、川芎增大；EC50较单用淫羊藿、川芎加和减小，说明淫羊藿-川芎配伍优于单用淫羊藿、川芎。研究成果为临床应用淫羊藿-川芎治疗OA提供科学依据和实验支撑，初步揭示了补肾活血药对淫羊藿-川芎治疗OA的配伍机制和物质基础，诠释“补肾活血”治则的科学内涵，为从本质修复软骨的新药研发提供依据。