基于“毒-效”整合分析的制天南星炮制“减毒增效”机理研究

Arisaematis Rhizoma (AR) specializes in “eliminating dampness and phlegm”, and is one of the most important drugs for treating damp-phlegm syndromes. Currently, the toxicological substances of AR is clear, however, active substances base and related mechanisms of “eliminating dampness and phlegm” and "toxicity-reducing and efficacy-maintaining" processing mechanism of AR are still not clear. Our preliminary experiment and literature analysis indicated that the toxicity of Arisaema in addition to toxic raphide irritation and secondary inflammation, and potential renal toxicity and dryness. At the same time, the water decoction of AR can significant down-regulate the MUC5AC which is a key target for release of phlegm. Therefore, this project is guided by the idea of the integrated analytical method of toxicity and efficacy, in order to dryness and renal toxicity of as AR toxicity evaluation index, and the therapeutic effect of damp-phlegm model rats as AR efficacy evaluation, effect integration strategy was used to integrate the toxicity and reduce phlegm effect of AR respectively. Take the semi-finished products in arisaematis rhizome preparatum processing link as research samples, the "spectral effect / poison" relationship and artificial neural network correlation analysis were used to research multivariate analysis the changes of complex substance in vivo and in vitro, to investigate the relationship between the material and the effect of Arisaema toxicodynamics. The results be conducive to elucidate "toxicity-reducing and efficacy-maintaining" processing mechanism of AR, and provides support for its rational clinical application.

制天南星功能“燥湿化痰”，为中医治疗“湿痰”的要药。目前制天南星毒性的物质基础及作用机理行业内已有深入研究，但其发挥功能的活性物质基础与制天南星炮制“减毒增效”机制尚不明确。通过前期研究和文献分析发现，天南星的毒性除了毒针晶刺激诱导炎症之外，还有潜在的肾毒性和燥性，同时制天南星水提物能明显抑制“湿痰”证模型大鼠痰液分泌量等，故本项目在毒性中药“毒效”整合分析的思路的指导下，以燥性和肾毒性作为毒性评价指标，以对湿痰模型大鼠的治疗指标作为药效评价指标，釆用效应整合策略对天南星炮制前后的毒性和化痰效应分别进行整合；把制天南星炮制过程各环节样品作为研究对象，采用“谱-效/毒”关系和人工神经网络物效关联性分析针对复杂效应物质的体内、外变化进行多元分析；探讨天南星毒-效物质与效应的相关性，阐明制天南星“减毒增效”的炮制机理，为其临床合理应用提供支撑。

本项目以天南星“燥性”为切入点，立足于提出的工作假说，主要进行了天南星毒性物质基础研究及炮制减毒研究、“燥湿化痰”物质基础研究及炮制增效研究、天南星毒效整合分析研究以及制天南星炮制过程化学成分变化的研究。. 根据大鼠肾毒性结合网络毒理学预测天南星的毒性成分可能为秋水仙碱，根据HK-2细胞KIM蛋白表达验证秋水仙碱的肾毒性。根据大鼠燥性评价结合系统溶剂法筛选燥性最强的部位为多糖部位。炮制后天南星中秋水仙碱含量降低，中性均一多糖的含量降低，可能是天南星炮制降低毒性的机制之一。. 根据高脂饮食下的过敏性哮喘大鼠的“湿痰”模型评价制天南星“燥湿化痰”效果强于生天南星。通过系统溶剂法筛选天南星的有效部位，多糖部位和正丁醇部位均能够降低NCI-H292细胞的炎症因子水平。通过网络药理学预测结合分子对接结果表明异夏佛塔苷是天南星治疗哮喘的主要成分，异夏佛塔苷也是正丁醇部位主要成分。炮制后天南星中异夏佛塔苷含量降低，总多糖含量增加，生姜多糖可能是天南星炮制增强疗效的机制之一。. 相比正常大鼠，生天南星和制天南星多糖均对“湿痰”模型大鼠表现出微弱的燥性，符合中医“有故无殒亦无殒也”的思想。同时生天南星和制天南星多糖均能减轻模型大鼠肺组织的病变情况，说明天南星多糖同时发挥“燥性”和“燥湿化痰”的作用。. 通过分析制天南星炮制不同环节的样品，秋水仙碱在水反复处理后样品中含量大大降低，在制天南星中未检测到。多糖在与辅料生姜一起处理的样品中含量增加，天南星均一多糖则含量下降。异夏佛塔苷随水浸泡时间越长损失越多，尤其随生姜加入损失增多。提示我们应该综合考虑制天南星的炮制工艺中水浸泡处理及辅料生姜对其品质的影响。. 基于以上分析，本课题组认为天南星的毒性和功效与其“燥性”有关，多糖、异夏佛塔苷、秋水仙碱都是其“毒-效”物质基础，制天南星炮制“减毒增效”的机制与天南星中性均一多糖、秋水仙碱含量降低和总多糖含量增加。.