基于细胞药代动力学与代谢组学的茵陈退黄机制与药效物质基础研究

The starting point of the project is to explore molecular mechanism and pharmacodynamic material basis of traditional Chinese medicine (TCM) for prevention and treatment of jaundice. Capillary Wormwood Herb was chosen as the research object. The human endogenous substance bilirubin was chosen as the biomarkers of jaundice in this study. The frontier methods and techniques of cellular pharmacokinetics and metabonomics, the primary hepatocytes, the "sandwich-cultured" hepatocytes, gene transfected MDCK-II cell models, and the transgenic animal model of jaundice were used to explore the influence of Capillary Wormwood Herb and its components on nuclear receptor PXR、CAR-regulated OATP1B1, UGT1A1,MRP2-mediated uptake, metabolism,efflux of bilirubin at the molecular, subcellular, cellular and animal levels, respectively. The effect of synergistic regulation from multiple components of Capillary Wormwood Herb on transport and metabolism of bilirubin based on multi-target was also observed. Using the four-dimensional research model “chemical fingerprint of TCM-pharmacokinetic markers-biomarkers-pharmacological targets”, the jaundice- relieving molecular mechanism and pharmacodynamic material basis of Capillary Wormwood Herb were investigated and interpreted from the perspective of regulation of nuclear receptors, transporters and metabolic enzyme. The research will demonstrate important theoretical value and clinical significance in disclosing the molecular mechanism and pharmacodynamic material basis of TCM in preventing and treating jaundice, and provide scientific basis and important reference for research, development and reasonable application of jaundice-relieving drugs as well as treatment of jaundice.

本课题从探寻中药防治黄疸的分子机制与药效物质基础科学问题出发，选择茵陈为研究对象，以人体内源性物质胆红素为主要生物标志物，采用细胞药代动力学和代谢组学前沿方法技术，构建原代肝细胞、“三明治培养”肝细胞、基因转染MDCKⅡ细胞及转基因黄疸疾病动物模型，从分子、亚细胞、细胞及整体动物水平上，研究茵陈及其组分对PXR、CAR等调控的OATP1B1、UGT1A1、MRP2等介导的胆红素摄取、代谢、排泄的影响；考察茵陈多组分对胆红素转运代谢的多靶点协同调控作用；采用“中药化学指纹-药代标示物-生物标志物-药效靶标”四维模式，从药动学和代谢组学独特视角，从核受体、转运体和代谢酶调控角度，研究并阐释茵陈退黄的分子机制与药效物质基础。本研究对于揭示中药退黄机制与药效物质基础具有重要理论价值和临床意义，为退黄药物研发、临床合理应用及黄疸治疗提供科学依据和重要参考。

本课题从探寻中药防治黄疸的分子机制与药效物质基础科学问题出发，选择茵陈及其主要组分为研究对象，以人体内源性物质胆红素为主要生物标志物，采用细胞药代动力学、代谢组学等前沿方法技术，构建了胆红素代谢指纹图谱、大鼠原代肝细胞、“三明治培养”原代肝细胞、OATP1B1基因转染MDCKⅡ细胞以及黄疸疾病动物模型，研究了茵陈及其主要组分对上游CAR、PXR、AhR和下游OATP1B1、UGT1A1、MRP2表达的影响，揭示其对CAR、PXR、AhR调控，人OATP1B1介导的胆红素摄取、人UGT1A1（肝微粒体）介导的胆红素葡萄糖醛酸结合以及MRP2介导的胆红素外排的影响。.研究结果表明，茵陈提取物及其所含的香豆素类（滨蒿内酯、东莨菪内酯、7-甲基香豆素）、黄酮类（茵陈色原酮、对羟基苯乙酮、异鼠李素3-O-葡萄糖苷）和有机酸类（绿原酸、隐绿原酸、新绿原酸、异绿原酸B和异绿原酸C）组分可显著增加胆红素摄取(0.93~9.79倍)、代谢（0.46~0.98倍）和外排（0.46~2.43倍）。这些组分是OATP1B1、UGT1A1、MRP2的潜在激活剂（可能不是CAR、PXR、AhR调控剂），是茵陈退黄的主要药效物质基础。茵陈退黄主要是由于其所含的香豆素类、黄酮类、有机酸类组分直接增强了OATP1B1、UGT1A1、MRP2基因和蛋白表达（CAR、PXR、AhR表达未发生明显改变），增加了这些转运体和代谢酶介导的胆红素肝细胞摄取、代谢和外排，促使胆红素从体内清除所致。.本课题采用“中药化学指纹-药代标示物-生物标志物-药效靶标”四维模式，从分子、基因、蛋白、细胞及整体动物水平上，从药动学和代谢组学独特视角，从核受体、转运体和代谢酶调控角度，研究并阐释了茵陈退黄的分子机制与药效物质基础。本研究对揭示中药退黄机制与药效物质基础具有重要理论价值和临床意义，为退黄药物研发、临床合理应用及黄疸精准治疗提供科学依据和重要参考。