傈僳族药金不换与乌头属药物的致毒/解毒生物学表观特征及机制研究

The aconitum medicine were used extensively and led to high death rate in clinical in our country. The detoxificated medicine available are not very wonderful. The present project has established an effective and mechanism- definited screening model for sodium channel of cell membranes using patch clamp technique. This model could be used to evaluated the toxicity of aconitum medicine and their detoxification. Veratrilla baillonii Franch, a folk medicine from Lisu nationality was confirmed to significantly antagonized the toxicity induced by aconitum alkaloids and worth investigation and exploitation deeply. Our study is planned to screening the active components of Veratrilla baillonii Franch from the whole body to molecular level. Using the mutiple methods and technique such as toxicological, whole cell patch clamp, gene chip and metabolomics, the interaction between Veratrilla baillonii Franch and aconitum plants as well as their apparent characteristics of poison/detocication are to evaluated objectively, the active targets for Veratrilla baillonii Franch are to be found, and lastly, the molecular mechanism and signal pathway of the medicine are to be clarified. The finish of the project will provide overall technical data for the reasonable utilization and deeply exploitation for Veratrilla baillonii Franch, supply new methods to save the drug poison of aconitum plants in clinical and settle theoretical fundament to derive new formula for aconitum medicine.

乌头属药物在我国大范围、大量使用，临床中毒率及死亡率极高，现有的解毒药物不完全理想。本项目在前期调查和研究基础上，应用膜片钳技术，建立了一种有效的、毒理机制相对明确的细胞膜钠离子通道快速筛查模型，用于评价乌头类药物毒性及解毒药作用，并证实傈僳族药物金不换（Veratrilla baillonii Franch）根茎提取物能够显著对抗乌头类生物碱所致中毒，是一种值得深入研究和开发的民族药。本研究拟从整体、组织、细胞、分子等多层面展开金不换解救乌头类中毒的活性成分筛查，采用毒理学、膜片钳、基因芯片、代谢组学等方法和技术客观评价金不换与乌头类的相互作用及致毒/解毒生物学表观特征，探寻金不换解救乌头类中毒的活性靶标，阐明其解毒分子机制及信号通路。本项目的完成，将为金不换资源的合理利用和深度开发提供全面技术资料，为解救乌头类药物中毒的研究提供新的思路，为乌头类药物衍化新方奠定理论基础。

项目围绕乌头类药物的毒作用及机制，采用多种实验技术手段，探讨金不换对乌头类药物急性、亚急性及体外细胞毒作用的致毒/解毒机制。明确了毒药的量-毒-效关系，并进一步研究其致毒/解毒作用、作用机制和物质基础。（1）系统提取分离其活性部位。得到35个化合物，利用现代波谱学知识并结合其理化性质已鉴定其中 26 个化合物的结构，包括18个酮类化合物，6个环烯醚萜类化合物，2个酯类化合物。其中发现新化合物1个，16个化合物为首次从本属植物分离。（2）证实了黄秦艽对雪上一枝蒿所致的急性、亚急性毒性均有缓解作用，其作用靶器官包括心、肝、肾等。可改善雪上一枝蒿所致心脏、脑组织代谢紊乱，缓解乌头类所致心肌细胞钠离子通道激活，并诱导肝脏CYP3A3和CYP2D6活性，促进乌头类代谢而解毒。（3）对其体内外药效进行了初步筛选，证实了黄秦艽中环烯醚萜苷类的保肝、抗氧化及肝药酶诱导作用。对其干预雪上一支蒿所致肝脏、心脏靶器官进行了基因芯片分析。并用信息生物学手段预测其活性分子的保肝作用，证实了其环烯醚萜苷类成分的肝保护及促进乌头碱代谢解毒作用。研究结果对寻找乌头类天然解毒剂、探讨其致毒/解毒生物学特征与机制具有重要意义。