基于“有故无殒”思想和TK-TD结合模型的栀子肝毒性“量-时-毒”关系及体内毒性物质基础研究

Gardenia is one of the first batch of Medicine and food　dual-use herbs announced by the Ministry of Health,commonly used in jaundice hepatitis treatment，however reported the presence of hepatoxicity. Hepaxicity or liver protection? What will happens to jaundice hepatitis animals(positive huang Syndrome)at the dose which induced hepatoxicity in normal animals? Does liver protection and hepatoxicity double effect exist in jaundice hepatitis animals? What is the conversion point of dose and time? What is the material basis of gardenia hepatoxicity? In Clinical how can use reasonablely?all of these are worth studying further.Not only there is few studies focus on the dose-time-toxic relationship of normal animals,but also no hepatoxicity research about jaundice hepatitis animals(positive huang Syndrome). The material basis of gardenia hepatoxicity is unclear.So our project will conduct the dose-time-toxic relationship research with normal animals and positive huang Syndrome animal based on .theory of YOU GU WU YUN,which fully reflects the real situation of gardenia in clinical ,and to determine the gardenia safe and effective" therapeutic window" and the course in animal level and elucidate dose-time change condition of liver protection and hepatoxicity．TK-TD reaearch will launch in normal and disease animal on geniposide and its metabolites ,to clarify influence by disease status on gardenia metabolic regularity, thereby conclude hepatoxicity material basis of Gardenia.This research provides the objective basis for gardenia rational regimens ( dose, duration ) in clinical treatment, at the same time promotes healthy development of Gardenia industry.

栀子长期广泛地被应用于治疗湿热黄疸，然而近年来其单独应用于正常动物体内却表现出明显的肝毒性。作为卫生部公布的首批药食两用中药，其安全性不容忽视。因此我们需明确以下问题：①导致正常动物产生肝毒性的剂量对阳黄证动物产生的是肝保护还是肝毒性？②栀子是否存在肝保护和肝毒性双向作用？如客观存在，其发生转换的剂量和时间节点是什么？③栀子产生肝毒性的体内物质基础是什么？目前尚未见上述问题的系统研究和报道。本课题在内经“有故无殒”思想指导下，独辟蹊径地同时采用正常和阳黄证动物模型系统开展“量-时-毒”关系研究，明确栀子肝保护和肝毒性双向作用的客观存在性及其转换的量时特征；从而确定栀子在正常和阳黄证动物体内的安全剂量和疗程，采用TK-TD结合模型明确栀子苷及其代谢产物在正常动物和病证动物体内的药动学特征差异以及栀子肝毒性的体内物质基础。为制订栀子临床安全合理给药方案、栀子肝毒性的监测和预警提供直接客观依据。

本课题在有故无殒理论的启发下，采用正常大鼠和黄疸模型大鼠对栀子的肝肾毒性及其体内毒性物质基础进行了较系统的研究，研究结果表明1）1.2g/kg-1栀子苷可对正常及黄疸模型造成明显的肝肾损伤，24h、48h为其肝肾毒性高峰，对黄疸模型造成的肾损伤较正常大鼠严重，240h后损伤基本自我修复。2）栀子提取物单次给药剂量超过1.16 g·kg-1(折算至临床用量为172g栀子生药）即可对正常大鼠产生急性肝毒性，栀子提取物单次给药剂量超过0.76g·kg-1(折算至临床用量为113g栀子生药）即可加重对黄疸模型大鼠的急性肝损伤；栀子提取物单次给药0.48 g·kg-1(折算至临床用量为71.5g栀子生药）即可导致正常及黄疸模型大鼠的急性肾损伤，1.16g·kg-1(折算至临床用量为172g栀子生药）剂量以上对黄疸模型大鼠的肾损伤较正常大鼠组更为严重。3）长期毒性量-毒关系研究表明栀子提取物剂量超过0.56g·kg-1(折算至临床用量为83.4g栀子生药）连续给药28d可对正常大鼠造成一定肝损伤，栀子提取物剂量超过0.28 g·kg-1(折算至临床用量为41.7g栀子生药）连续给药14d和0.14g·kg-1(折算至临床用量为20.8g栀子生药）连续给药14d即可加重黄疸模型大鼠的肝损伤；栀子提取物剂量超过0.56g·kg-1连续给药14d可对正常大鼠造成一定肾损伤，栀子提取物剂量超0.28 mg·kg-1连续给药14d即可对黄疸模型大鼠造成肾损伤；且停药两周后栀子提取物剂量在0.14~0.56g·kg-1对大鼠造成的肝肾损伤是可逆的。4）TK-TD研究表明栀子苷为栀子产生肝毒性的物质基础，栀子加重黄疸模型肝肾损伤的原因可能与黄疸模型大鼠体内栀子苷的CMAX、AUC明显增大,MRT和t1/2z延长，CLz/F降低有关。5）京尼平可能为导致栀子产生肝肾毒性的体内毒性物质基础。本课题的实施初步阐明了栀子肝肾毒性的量-时-毒关系及体内毒性物质基础，为栀子的临床安全合理用药方案的制定及其肝肾毒性的预警和监测提供了直接的科学依据。