基于炮制减毒思想的何首乌肝毒性物质基础初步研究

Preliminary study on hepatotoxic components in Polygoni Multiflori Radix based on processing and toxicity-decreasing

目的基于何首乌炮制前后的谱-毒相关分析,筛选何首乌肝毒性物质基础,为提高何首乌药材质量控制方法,确保临床用药安全提供参考。方法以何首乌生品及黑豆汁蒸制不同时间的炮制品为研究对象,采用UPLC-Q/TOF-MS技术表征各样品的化学信息,并结合文献初步指认其主要成分;再以正常人肝细胞(Lo2细胞系)为模型,细胞抑制率为评价指标,采用简单相关分析及多元线性相关分析方法,筛选何首乌致肝毒性的主要成分。结果共指认出何首乌生品及炮制品中的7种主要共有成分反式二苯乙烯苷、没食子酸、大黄素、大黄素甲醚、大黄素-8-O-β-D-葡萄糖苷、顺式二苯乙烯苷、儿茶素,并基于谱-效简单相关分析发现反式二苯乙烯苷、大黄素甲醚、大黄素-8-O-β-D-葡萄糖苷、顺式二苯乙烯苷、儿茶素5个成分与何首乌毒性相关性较强。进一步主成分回归分析发现大黄素甲醚与顺式二苯乙烯苷对何首乌毒性贡献度较大,提示这2个成分可能是何首乌主要毒性成分。何首乌高压黑豆汁蒸至36 h后才能达到减毒的效果。结论该研究可为何首乌的合理利用和毒性研究提供参考和依据。

Objective To explore the main toxic components induced hepatotoxicity based on the spectrum-toxicity correlation analysis of processed Polygoni Mult^ori Radix （PMR）, and to provide reference of PMR for promoting the quality control as well as the safe clinical treatment. Methods The UPLC-Q-TOF-MS characterized chemical information of all samples including crude and processed PMR which were black soybean steamed with high pleasure in different time, and the main components were identified after referring to the literatures. With the normal human hepatocytes （Lo2 cell line） as in vitro model and cell inhibitory rate as testing index, the simple correlation analysis and multivariate linear correlation analysis were used to screen the main components of PMR hepatotoxicity. Result Seven main same components of crude and processed PMR were identified as trans-stilbene glycoside, gallic acid, emodin, physcion, emodin-8-O-β-D-glucoside, cis-stilbene glycoside, and catechin. And it was found that trans-stilbene glycoside, physcion, emodin-8-O-β-D-glucoside, cis-stilbene glycoside, and catechin were closely related to PMR hepatotoxicity. Furthermore, physcion and cis-stilbene glycoside were found greatly contributing to hepatotoxicity induced by PMR in the principle component regression analysis, which indicated these two might be the main toxic components. In addition, the results demonstrated apparent effects in detoxification could not be achieved until black soybean steamed with high pleasure at least 36 h. Conclusion This study will provide data support for further rational use of PMR and deep hepatotoxicity research.