关木通等含马兜铃酸中药材及中成药毒理机制的代谢组学研究

目的: 研究含马兜铃酸药材及中成药给药大鼠后内源性代谢产物的动态变化过程,表征其毒性反应特征性代谢谱,探讨其毒理作用机制。方法: 将大鼠分组,分别灌胃给药马兜铃酸Ⅰ对照品与药材(关木通、马兜铃、青木香和天仙藤)和传统方剂(龙胆泻肝丸和导赤丸)的提取物。采集血浆和尿样进行气相色谱-飞行时间质谱(GC-TOF/MS)测定和多元数据分析,临床生化及组织病理检查。结果: 受试大鼠出现急性毒性反应,血浆与尿样中柠檬酸、甘氨酸、丝氨酸、乌头酸、棕榈酸、己二酸和对羟基苯乙酸等多种代谢产物发生显著改变,其变化水平与给药剂量和天数显著相关,各组间代谢谱差异可清晰区分机体不同染毒状态。马兜铃酸毒性作用除了导致肾小管受损,重吸收功能降低外,还与机体肠道菌群失调,代谢功能改变及三羧酸循环改变等多种环节密切相关。结论: 本实验采用基于GC-TOF/MS测定和多元数据分析的代谢组学研究,以内源性代谢产物谱表型表征机体染毒状态,解读含马兜铃酸药材及中成药的毒性作用与机制,对该类药材及中成药的安全性做出进一步的综合评价与探索。

Objective: To investigate Chinese medicinal herbs containing aristolochic acid and its prescriptions including Caulis Clematidis Armandii, Fructus Aristolochiae, Radix Aristolochiae, Herba Aristolochiae, Longdan Xiegan Wan, Daochi Wan by a global metabolic profiling of endogenous metabolites in rats, so as to understand toxicological mechanism of aristolochic acid. Methods: Plasma and urine samples were collected and analyzed by gas chromatography-time of flight mass spectrometry(GC-TOF/MS) technique integrated with pattern recognition. Serum biochemistry and histopathological examination were also performed. Results: Multivariate data analysis showed the time-related and dose-related metabolic trajectories induced by aristolochic acid. Aberrant levels of citric acid, aconitic acid, hexadecanoic acid, glycine, serine, hexanedioic acid and hydroxyphenylacetic acid were observed in body fluids, which suggested that toxicity of aristolochic acid had caused proximal convoluted tubule injury and decreased tubular reabsorption, as well as dysregulation of TCA circle and intestinal flora. Conclusion: The GC-TOF/MS-based metabolomic study provided a holistic approach for investigating the time-related and dose-related toxicity of aristolochic acid, and could be a comprehensive strategy for toxicology study and drug safety assessment of traditional Chinese medicine.