基于维吾尔医治疗肝病方剂配伍的菊苣作用的药效物质基础研究

Prescription of TCM or ethnomedicine were complicated chemical systems, its synergy and material basis were very important for curing diseases. Couplet medicines were the simplest synergy and prescription. Chemical components entered into blood can express the effects of prescription, exactly. Based on Serum Pharmacochemistry, Pharmacology, Pharmacodynamics and Proteomics techniques, HPLC-MS fingerprint chromatography of couplet seed-root of Cichorii glandulosum was established for analysis of synergy ratio and different decocting ways, in vitro. Investigated structures of components and metabolites in drug-containing serum by HPLC-MS. In addition, proliferation inhibition and apoptosis of hepatic stellate cell acted by drug-containing serum were assayed. MALID-TOF-MS based on proteomics were applied to research the differential expression proteins in hapatic injury rats, with the help of network pharmacology and multivariate statistical analysis to elucidate material and mechanism of the couplet, and screen the potential target of hepatic protection. In the end, material basis and synergy rules of Uygur hepatic protection prescriptions were evaluated, comprehensively. The implementation of the project provides experimental and theoretical basis for the exploration of the treatment of liver diseases and the deeper development of the Uygur classical liver protection prescriptions.

中药（民族药）方剂是一个复杂的化学体系，对其配伍规律和物质基础的认识是阐述方剂治病科学性的重要依据，而药对是最简的配伍。基于方剂中进入机体血液的物质能够更准确地表达其药效，本项目将血清药物化学、血清药理学、药效学、蛋白质组学相结合，采用HPLC/MS联用技术，建立维吾尔保肝经典方中菊苣子-菊苣根药对的指纹图谱，分析配伍比例和煎煮形式对化学成分的影响；确定吸收进入肝损伤大鼠体内的化学成分及其代谢物，研究该药对含药血清对肝星状细胞增殖和凋亡影响；基于蛋白质组学技术采用MALDI-TOF/MS等方法分别研究该药对对肝星状细胞和肝损伤大鼠肝组织差异表达蛋白，综合生物信息学、网络药理学及多元统计分析技术，力求系统阐明菊苣子-菊苣根药对肝保护作用的物质基础和作用机制，明确作用靶点，评价维吾尔医保肝方剂中菊苣的药效物质基础，为探索肝病治疗及深层次开发维吾尔医保肝方剂提供实验和理论依据。

中药（民族药）方剂是一个复杂的化学体系，对其配伍规律和物质基础的认识是阐述方剂治病科学性的重要依据，而药对是最简的配伍。复方配伍是维吾尔医药学理论的重要组成部分，在传统维吾尔药复方中，有一类配伍尤其特别，即同一药材的不同药用部位（称为“药对”）常常在不同处方中按不同比例配伍使用，以达到提高其疗效的目的。项目为探究新疆保肝经典药对菊苣根-菊苣子配伍的科学性，首先体外实验结果证实菊苣根-菊苣子配伍后的肝保护作用优于单味药材，如对肝星状细胞增殖的活化抑制作用和对肝癌细胞的抑制作用增强；其次采用指纹图谱结合UPLC/MS联用技术对药对配伍前后的化学成分进行定性定量分析，发现配伍后化学成分发生了苷键水解、酯水解等变化，形成更稳定、活性更强的化合物。进一步经谱效关系结合数学分析研究筛选出配伍后5个差异化合物具有显著的抗肝纤维化活性。利用血清药理学和血清药物化学技术发现配伍后大鼠含药血清具有较强的抗肝纤维化活性，活性可能与筛选出的20个差异化合物有关，其中11个为原型成分，9个为非原型成分。最后在动物肝纤维化模型中亦证实配伍后药对的抗肝纤维化作用强于单味药材，大鼠肝组织蛋白质组学研究显示：差异表达蛋白涉及的信号通路主要有GMP-PKG signaling pathway、ECM-receptor interaction、drug metabolism-cytochrome P450等，并对Galectin-1、Galectin-3、Glutamine synthetase和Itga2b等蛋白表达进行验证。本项目综合利用动物实验、细胞实验、分子实验、分析实验，结合药材指纹图谱技术、血清药理学、血清药物化学、蛋白质组学等相关技术，系统阐明了菊苣根-菊苣子药对肝保护作用的物质基础和作用机制，为探索肝病治疗及深层次开发维吾尔医保肝方剂提供实验和理论依据。