柴胡肝毒性与功效、证候关系的基础研究

Radix Bupleuri(RB) were listed as top grade in 《Shen Nong Ben Cao Jing》and has the effect of Shugan jieyu. Its toxic and side effects are generally recognized and was recorded as "Bupleurum robbing hepatic yin". A large-scale of toxic events of RB in Japan were considered that its hepatoxicity was related to protoplasmic poison. Saikosaponin (SS) and volatile oil are both the main active substances and toxic substances of RB. The expression of hepatoxicity in rats and mice, "dose-time-toxicity" relationship, infected path, response mechanism in cell, molecule and gene level, and the expression of differential proteins of T-SS were found by the previous researches. According to all of these, we raise the presumption that "Toxicity of RB is closely related to its efficiency and symptom, and this can be cognized and represented scientifically. This is based on the premise of scientific control and secure use of RB in the theory of traditional Chinese medicine." Correlation studies of the relationship between pharmacological activity and hepatoxicity of RB should be performed both in vivo and In vitro in the level of entirety, organ, cell and molecule. Pharmacology and interconnected characteristics of RB's efficiency and hepatoxicity, plus its mechanism should be cleared, then to verify it based on symptoms. This aim at finding out the mechanism and characteristics of "Toxicity-Efficiency-Symptom" of RB, and to establish modern represented systems, scientific recognized models and secure drug use strategies of RB's hepatoxicity based on its efficiency and symptom.

柴胡为《本经》中上品，具"疏肝解郁"之功，后世有"柴胡劫肝阴"之载，尤以日本大规模"柴胡中毒事件"为甚，认为肝毒性与原浆毒有关。柴胡皂苷（SS）、挥发油是主要化学和药效物质，前期研究证实T-SS和挥发油是主要肝毒物质，已获知大、小鼠体内肝毒性表现、"量-时-毒关系"、染毒路径及细胞、分子、基因水平应答机制和T-SS基于"量-时-毒" 关系的差异蛋白表达。据此，提出"柴胡肝毒性与功效、证候密切相关，在此背景下可被科学认知和正确表征，这是基于中医理论的科学控制和安全使用柴胡的前提"的工作假说；运用体内、外研究手段在整体、器官、细胞、分子水平上开展柴胡药理活性与肝毒性相关性研究，明确柴胡发挥功效、表达肝毒性的药理学特征、体内关联特性和内在机制；并进行证候背景下验证。旨在解决柴胡"毒性-功效-证候"发生机制和关联特性，构建柴胡肝毒性在功效与证候背景下的现代表征体系、科学认知模式和安全用药策略。

柴胡为《本经》中上品，具"疏肝解郁"之功，后世有"柴胡劫肝阴"之载，前期研究证实柴胡总皂苷和挥发油是主要肝毒物质，获知其体内肝毒表现 、"量-时-毒关系"、染毒路径及细胞、分子、基因水平应答机制和T-SS基于"量-时-毒" 关系的差异蛋白表达。据此，本课题提出“柴胡肝毒性与功效、证候密切相关，在此背景下可被科学认知和正确表征，这是基于中医理论的科学控制和正确使用柴胡的前提”的工作假说。.在文献研究基础上，从毒性记载与功效和证候背景相关的角度，进一步厘清柴胡“疏肝解郁”功效和“柴胡劫肝阴”等毒性记载的历史文化背景、发生模式及不同时代的异同点和发展演变规律；运用体内、外多种实验手段和多学科交叉研究，在整体、器官、细胞、分子水平上系统开展柴胡不同剂量水平的镇静、解热、抗癫痫、抗癌、抗肝纤维化等药效学研究，进行药效的量-效关系研究和相关作用机制探讨，明确发挥药效作用的有效剂量范围。获知了上述柴胡样品发挥药效的有效剂量下出现的伴随毒副作用，包括一般状况、生理指标、肝脏功能、形态改变来明确柴胡发挥功效和产生肝毒性的物质基础、药理学性质以及作用特点是否一致，得悉其“有效治疗窗”和“药物安全窗”，明晰柴胡发挥“疏肝解郁”功效以及“柴胡劫肝阴”毒性的物质基础、药理学特点、作用路径的异同，并以柴胡为例进一步核实中药多成分、非单一效应的特点，回答了“中药成分有毒≠部位有毒≠组分有毒≠单味药有毒≠中药复方有毒”的问题，通过不同病证背景下验证，从剂量范围、作用特点等方面探讨柴胡“毒性-功效-证候”发生机制和关联特性，构建柴胡肝毒性在功效与证候背景下的现代表征体系和科学认知标准，为临床提供柴胡安全、合理应用的最佳剂量和进一步开发基于柴胡传统功效和现代研究的安全、高效、利于吸收的创新药物打下坚实的基础。以本项目为基础申报的“中药肝毒性早期发现与合理评价技术平台建设及其科学应用”获得2015年度山东省科技进步一等奖。