基于药物代谢组学的中药肉苁蓉抗血管性痴呆体内药效物质及作用机制研究

Vascular dementia (VD) is caused by problems in the supply of blood to the brain, typically by a series of minor strokes. The term refers to a group of syndromes caused by different mechanisms all resulting in vascular lesions in the brain and Chinese medical theory regarded that this disease was caused by kidney essence deficiency. Cistanche Herba (CH) is one of the most famous tonic herbal medicines. As an effective fraction preparation, CH total glycosides capsules (Memoregain) have been widely used in clinic for the treatment of vascular dementia; however, the efficacious material basis as well as the effective mechanism hasn’t been revealed, which is a great obstacle for its further development. In this project, pharmaco-metabonomics, which is a special metabonomics and has been widely adopted to study the therapeutic mechanism of medicine via monitoring drug-derived xenobiotics at the meanwhile of endogenous metabolome profiling, is introduced aiming to clarify the material basis along with the mechanism for the anti-dementia effect of CH. The whole strategy can be divided into a couple of steps: first, VD animal model is going to be developed; second, we will characterize the metabolic profiles of representative compounds using in vivo and in vitro metabolic models and characterize the generation of the in vivo CH-related components; third, multivariate statistical analysis and multiple strategies, such as Lightsight software, MetaboLynx software, mass defect filter (MDF) and chemicalome and metabolome matching approach (CMMA) strategies, will be adopted to find the CH-derived components in vivo; fourth, the metabolites will be purified using in vitro incubation, biological sample-based isolation and wet-chemistry based method; fifth, the activities of the isolated metabolites will be screened using pharmacological models; sixth, the endogenous metabolome will be adopted as the therapeutic indicators to construct the pharmacokinetics-pharmacodynamics (PK-PD) model with the multi-components pharmacokinetics of CH; and seventh, the correlation between CH-derived efficacy substances and their related endogenous metabolic pathways will be characterized to determine the therapeutic mechanism of CH. The findings obtained in this project are expected to provide important information for the characterization of efficacy material basis and mechanism for traditional Chinese medicine using pharmaco-metabonomics based technique roadmap.

血管性痴呆是指由各种脑血管疾病而导致的脑功能障碍从而产生大脑智能及认知功能障碍的临床综合征，是老年期痴呆的常见病因之一，其发病机制中医认为由肾精亏虚。肉苁蓉为著名的补益类中药，其有效部位制剂苁蓉总苷胶囊在临床上被广泛用于血管性痴呆的治疗，但体内药效物质及作用机理尚未明确。药物代谢组学通过同时检测药物体内成分与其引起的代谢轮廓变化来阐明药物的作用机理。本项目引入药物代谢组学的方法，对肉苁蓉的体内药效物质及其作用机理进行深入的研究。以血管性痴呆模型动物为生物体系，结合多种质谱技术及策略鉴定体内药源性成分，对体内代谢产物进行提取分离和活性筛选，以内源性代谢轮廓为体内药效指标之一，结合其他药效指标，通过构建肉苁蓉多成分药动学及内源性代谢轮廓的药动-药效模型，进行生物学分析，阐明肉苁蓉防治血管性痴呆的体内药效物质及作用机制。本项目将为基于药物代谢组学中药体内药效物质及作用机理的研究提供参考。

血管性痴呆是指由各种脑血管疾病而导致的脑功能障碍从而产生大脑智能及认知功能障碍的临床综合征，是老年期痴呆的常见病因之一，其发病机制中医认为由肾精亏虚。肉苁蓉为著名的补益类中药，其有效部位制剂苁蓉总苷胶囊在临床上被广泛用于血管性痴呆的治疗，但体内药效物质及作用机理尚未明确。本项目以药物代谢组学为主要研究方法，以阐明肉苁蓉防治血管性痴呆的体内药效物质及作用机制为研究目标。从2015年1月至2017年12月，课题组分别从化学成分分离、肉苁蓉化学成分组和生物样品代谢物组动态表征、药物代谢、代谢组学和药物代谢组学等方面系统地开展了研究工作。获得化合物26个，其中苯乙醇苷类化合物18个；利用LC-MS技术从管花肉苁蓉和荒漠肉苁蓉中共发现并鉴定513个化学成分，并比较了两者化学成分谱的差异；分别建立了快速和全面分析两种肉苁蓉质量分析的方法；建立了生物样品代谢物组定量表征策略；深入阐明了肉苁蓉给药后体内代谢产物；通过给药后生物样品中内源性代谢物组和外源性代谢物组的动态表征，构建了药源性成分和病理生物标志物之间的动态关系。系列研究结果表明肉苁蓉改善血管性痴呆的主要活性形式为苯乙醇苷类化合物的代谢产物群，如羟基酪醇磺酸酯、甲基羟基酪醇磺酸酯、羟基酪醇糖醛酸酯、高香草酸、高香草酸磺酸酯、高香草酸糖醛酸酯等多个苯乙醇苷类化合物的体内代谢产物，而起效机制主要为直接纠正酪氨酸及色氨酸代谢途径、补充神经递质、清除氧自由基等，从而治疗血管性痴呆。发表期刊论文22篇，其中SCI 20篇，第一作者或通讯作者20篇。在分析化学领域顶级期刊Analytica Chimica Acta发表研究性文章2篇、色谱领域顶级期刊Journal of Chromatography A发表研究性文章7篇。获得国家科技进步二等奖和高等学校科研究优秀成果奖(推广类)一等奖各一项。培养科研骨干2名，博士1名，硕士2名。较好地完成了项目预期目标，所获得的成果具有较高的学术价值和应用前景。