基于“效应成分集成致毒”假说的栀子厚朴汤潜在毒性机制研究

The research of basis of toxic substances of “prescriptions with potential toxicity” should not only consider the integration effect of active ingredients after compatibility of prescription in the process of ADME, but also give consideration to their induction in toxicity production. Under the “toxic dose” and “toxic condition” of Zhi-Zi-Hou-Po decoction, qualitatively and quantitatively studied the absorption, distribution, metabolism, and excretion of chemical constituents in the animal body. By examining the relationship between the dose-exposure state-injury degrees, expound the correlation between the exposure and toxicity of Zhi-Zi-Hou-Po decoction from the level of in vivo effect substances and concentrations. At the same time, metabolomics is used to study the end product of the whole metabolic network of the organism and quickly discovered the toxicological regularity of the Zhi-Zi-Hou-Po decoction. Comprehensive assess the combined toxicity of multiple targets from multiple perspectives. Combine network pharmacology, explore the metabolic network of toxic damage, and establish a background network of effect components based on database information. By analyzing the topological correlation between the damage metabolic network and the compound background network, the in vivo components of Zhi-Zi-Hou-Po decoction are linked to the toxic mechanism. This study reveals the toxic substance basis and possible poisoning mechanism of Zhi-Zi-Hou-Po decoction by explore the dynamic changes of the effector network and injury pathway and non-dominant toxic components.

“潜在毒性方剂”的致毒物质基础研究既要考虑方剂配伍后活性成分在ADME过程中的集成效应，同时还应兼顾它们在毒性产生中的诱导作用。定性定量地研究栀子厚朴汤在“显毒剂量”及“显毒条件”下，化学成分群在动物体内的吸收、分布、代谢、排泄过程，通过探讨给药剂量-暴露状态-损伤程度之间关系，从体内效应物质及浓度水平上阐明栀子厚朴汤体内成分暴露量和毒性的相关性。同时，采用代谢组学研究机体整个代谢网络的终端产物，快速发现栀子厚朴汤出现毒性的规律，从不同角度对多靶点的综合毒性反应进行全面评价，结合网络药理学，探究毒性损伤代谢网络，依据数据库信息建立效应成分背景网络，通过分析损伤代谢网络与效应成分背景网络的拓扑相关性，将栀子厚朴汤体内成分与致毒机制相关联。探讨效应网络-损伤途径-非显性毒性成分群的动态变化，揭示栀子厚朴汤毒效物质基础及可能致毒机制。

抑郁症具有病程长、复发风险高的特点，长期用药是其治疗的特点。栀子厚朴汤（ZZHPD）由“无毒药材”组成，用于抑郁症治疗。对大鼠连续给药，观察到3倍及以上临床剂量均有肝肾损伤，且肝毒更为明显，损伤程度与剂量、时长正相关，提示其用于抑郁症治疗存在安全风险。本研究在对ZZHPD有较为深入探究的基础上，开展代谢组学及网络药理学研究，追踪大鼠毒性发生、发展过程中肝、肾损伤相关的生物标记物及其变化，探究损伤导致代谢网络变化的进程，揭示ZZHPD致肝、肾损伤的内在 “调控机制”。对给药ZZHPD大鼠血浆、尿液、肝脏代谢组学研究鉴定出的差异性代谢物进行分析,发现ZZHPD对氨基酸类、脂质类、核苷类、胆汁酸类等多种内源性代谢物水平均产生了干扰。其中，血浆样本代谢组学差异代谢物侧重于氨基酸类，尿液样本侧重于小分子有机酸类，而肝脏样本则侧重于胆汁酸类、肽类。通过ZZHPD给药后体内过程分析，采用系统识别策略，鉴定出汤剂入体成分。对给药后外源性成分栀子苷在血浆、组织（心脏、肝脏、脾脏、肺、肾脏和大脑）暴露量变化的研究，发现了栀子苷产生毒性机制。采用“代谢组学辅助网络药理学”研究策略，通过相关数据库搜索ZZHPD入体成分相关靶点，查找代谢组学研究发现的损伤代谢网络靶点，作为潜在毒性靶标,构建“潜在毒性靶标-成分相关靶标-成分-药材”拓扑网络，发现栀子苷、柚皮苷、橙皮苷、新橙皮苷、新圣草次苷、厚朴酚、和厚朴酚等8个与ZZHPD潜在毒性密切相关化学成分并加以确认。结果表明，栀子苷是栀子厚朴潜在毒性的关键成分，其它成分对栀子苷及其代谢物京尼平的毒性发挥起到了协同作用。本研究建立一种以非靶标代谢组学为评价手段， 以内源性及外源性代谢物系统辨识及动态变化为依据，结合病理、 生化研究，揭示“无毒中药”致损伤的研究体系。研究工作不仅为ZZHPD临床安全用药提供了科学依据，亦为中药潜在毒性物质研究提供了新的思路。