以“有故无殒”理论结合方症-时量-效应关系研究配伍大黄的肝肾毒性

Rhubarb, the Chinese herbal, is widely uesd to the prescription compatibility. In China, more than one tenth of Chinese medicine prescriptions are composed of Rhubarb. The liver and kidney toxicity have been reported by scientists from both domestic and overseas. In China, interestingly, following the theory of “You Gu Wu Yun”, the injury incident of liver/kidney was not found. However, the potential hepatotoxicity/nephrotoxicity of Rhubarb which is integrated in the prescription compatibility are still concerned. Hence, for this project, we employ the classical prescription, Yin Chen Hao Tang, to remedy the corresponding jaundice symptom as the research paradigm. Firstly, The pharmacokinetic(PK) parameters of multi-components of rhubarb which is applied to the prescription compatibility, namely, in Yin Chen Hao Tang, and were carried to treat the jaundice animal will be integrated with the analysis results of pharmacodynamics(PD) upon the liver and kidney; secondly, the effective sample collection time-point for the toxicity and metabolomics analysis are be made according to the PK-PD , and then the datum of toxicity analysis and metabolomics are acquired; thirdly, the analysis results of PK, PD, and metabolomics are correlated the Formula Symptom. The analysis technical system which be constructed by integrated with Formula Symptom, Time Quantities, as well as Effect will be used to analyze the toxicity of Chinese herbal medicine. Based on this, the jaundice model animals which are treated by Yin Chen Hao Tang are deeply explored in both hepatotoxicity and nephrotoxicity effect. Based on it, we will provide more reasonable theory for the rhubarb prescription compatibility, and the much more sound analysis technics system will be supplied for assessment the safety of rhubarb prescription compatibility; the more comprehensively toxicity analysis technics platform will be set up for analysis the toxicity of compound compatibility by virtue of this project.

中药大黄在方剂配伍中应用极其广泛，国内有超过1/10的复方含有大黄。国内外研究发现单用大黄具有肝肾损伤现象；但临床上依“有故无殒”理论，将大黄配伍使用后并未出现大量肝肾损伤事件。然而人们依然担忧配伍大黄后的潜在肝肾毒性。基于此，本项目以茵陈蒿汤治疗黄疸模型为范例。首次将复方中大黄多成分代谢动力学参数与肝/肾脏药效学分析结果拟合，筛选合理的药效学和毒性代谢组学采集时间点，关联方症、药动、药效、代谢组学结果，建立基于方症-时量-效应的中药复方毒性研究技术体系。依此技术体系，对茵陈蒿汤作用的黄疸模型进行两方面的深层探索：一、探究复方大黄在黄疸模型中对肝脏(病理组织)的潜在毒性；二、探究复方大黄在黄疸模型中对肾脏(非病理组织)的潜在毒性。以此为中药大黄的复方毒性研究提供有利的技术支持，为中药大黄的合理配伍使用提供进一步的理论支撑，通过本项目的研究搭建更全面的复方毒性技术研究平台。

有毒中药的毒-效特性是人们关注的重点。毒性中药大黄在临床肝病治疗上有着广泛的应用，其治疗肝损伤的“有故无殒”机理和潜在肝肾毒性特点有待深入研究。 .本项目采用代谢组学结合生化病理指标变化研究大黄单味药，大黄栀子药对，大黄栀子茵陈蒿复方在正常大鼠和病理模型大鼠的内源性代谢变化特征。通过研究发现大黄、大黄栀子、茵陈蒿全方对黄疸模型大鼠的体内毒-效作用状态。首次发现了中药诱导生物标志物变化的“有故无殒”特性，毒性药物通过调节内源性代谢产物变化，进而转变给变代谢通路发挥潜在的治疗和毒性特征，阐明有毒中药的潜在可用性和毒害性。获得大黄调控病理模型“有故无殒”效应标志物49个，改善代谢通路20条 ；毒性调控标志物18个，损伤代谢通路6条；大黄栀子调控病理模型“有故无殒”效应标志物61个，改善通路30条；毒性标志物18个，损伤代谢通路30条；茵陈蒿汤全方调控“有故无殒”效应标志物31个，改善通路是31条，毒性标志物21个，损伤通路26条。结合生化病理学分析得出大黄药量为15mg.100g-1连续给药一周能造成肝肾脏代谢扰动；大黄栀子配对(大黄药量45mg.100g-1)时连续给药一周能够造成显著肝肾损伤；在茵陈蒿汤全方配伍(大黄药量45mg.100g-1)时连续给药一周能够显著造成肝肾损伤。本研究从代谢层面诠释了有毒中药大黄在对应病理症候使用时确实存在潜在肝肾代谢扰动，其作用机制是通过调控特定内源性代谢产物发挥各自的药效与毒性特征，为了安全使用应该密切观察肝肾功能不全病人的用药或者进一步研究找到关联毒性成分组加以剔除。 .本研究为诠释有毒中药大黄应用的“有故无殒”理论提供了充分的科学证据，对基于“有故无殒”原则的配伍中药毒性研究提供了代谢层面的理论参考和技术支持。