4-烯丙基苯甲醚代谢活化及其代谢物与谷胱甘肽共价结合  

Glutathione conjugation resulting from metabolic activation of estragole

作　　者：周梦悦 李靖 程婷[1,2] 江婷婷 谭荣 陈妍 李维维 郑江 ZHOU Mengyue;LI Jing;CHENG Ting;JIANG Tingting;TAN Rong;CHEN Yan;LI Weiwei;ZHENG Jiang(School of Pharmacy,Guizhou Medical University,Guiyang 550004,China;Guizhou Provincial Key Laboratory of Pharmaceutics,Guizhou Medical University,Guiyang 550004,China;Wuya College of Innovation,Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]贵州医科大学药学院,贵州贵阳550004 [2]贵州医科大学贵州省药物制剂重点实验室,贵州贵阳550004 [3]沈阳药科大学无涯创新学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2023年第9期1173-1179,共7页Journal of Shenyang Pharmaceutical University

摘　　要：目的 研究4-烯丙基苯甲醚(estragole, ETG)的反应性代谢产物与谷胱甘肽(glutathione, GSH)在体内外形成的共价结合物。方法 采用高效液相色谱-三重四级杆串联质谱联用仪(LC-MS/MS)建立测定其代谢物与谷胱甘肽形成的相应结合物的液质分析方法,检测化学合成、体外小鼠肝微粒体孵育及体内小鼠肝脏中ETG反应性代谢产物与GSH形成的结合物。结果 在小鼠肝微粒体孵育液中采用GSH作为捕获剂,检测到了ETG反应性代谢产物与GSH形成的结合物(ETG reactive metabolites-GSH,ERM-GSH),该结合物具有α,β-不饱和醛结构,其色谱、质谱行为与化学合成样品一致;不同浓度1-氨基苯并三唑(1-aminobenzotriazole, ABT)抑制P450酶活性后,结合物的生成量随ABT浓度的增加而减少;在灌胃小鼠ETG的肝脏中观察到ERM-GSH结合物的生成量呈时间、剂量依赖性。结论 首次发现ETG代谢活化形成的α,β-不饱和醛反应性代谢产物在体内外均可与GSH反应形成ERM-GSH结合物,提示该代谢产物可能与蛋白质共价结合。Objective To investigate GSH conjugation resulting from metabolic activation of estragole(ETG)in vitro and in vivo.Methods Biomimetic synthesis of ERM-GSH conjugates was carried out by reaction of 4-methoxycinnamaldehyde with glutathione(GSH).ETG reactive metabolites-GSH conjugates(ERM-GSH)were determined using mouse liver microsomal incubation and animal models.Metabolites were analyzed by LC-MS/MS.Results An ERM-GSH conjugate derived from anα,β-unsaturated aldehyde was detected in liver microsomes fortified with GSH as a trapping agent which showed similar chromatographic and mass spectrometric behaviors as the synthetic conjugate.The inhibition of P450 enzyme activity by 1-aminobenzotriazole attenuated the formation of ERM-conjugate in concentration-dependent manner.The conjugate was also observed in liver of mice in a time-dependent/dose-dependent manner by intragastric administration of ETG.Conclusion ETG is metabolized to anα,β-unsaturated aldehyde reactive metabolite in vitro and in vivo.The resulting metabolite conjugates with GSH to produce an ERM-GSH conjugate,which suggests that the reactive metabolite could adduct hepatic protein.

关 键 词：4-烯丙基苯甲醚 谷胱甘肽结合物 代谢活化 

分 类 号：R917[医药卫生—药物分析学]