摘要
目的通过对槲皮素在人结肠腺癌细胞(Caco-2)中的代谢产物分析,探讨槲皮素在跨膜转运过程中的代谢特征。方法利用Caco-2单层细胞模拟人肠吸收模型,采用薄层色谱技术对槲皮素在Caco-2单层细胞模型上的代谢产物进行分离富集,并采用LC-MS和LC-MS/MS技术进行定性分析,以揭示槲皮素在肠吸收模型Caco-2细胞上基本的代谢转化特征。结果对槲皮素孵育Caco-2细胞裂解液和空白PBS孵育细胞裂解液进行LC-MS全扫描及对比分析,应用薄层色谱技术对可能的代谢产物进行分离富集,对薄层色谱分离富集的样品进行流动注射扫描分析。初步探明,槲皮素孵育细胞中其他可能的代谢产物尚有:槲皮素一硫酸酯、槲皮素三硫酸酯、槲皮素葡萄糖醛酸苷、槲皮素葡糖醛酸化硫酸酯、甲基槲皮素硫酸酯和甲基槲皮素葡萄糖醛酸苷,未观察到m/z>600的可能代谢产物。结论槲皮素在Caco-2单层细胞模拟的跨膜转运过程中伴随着广泛的代谢转化,主要以甲基化、硫酸化和葡萄糖醛酸化为主,未观察到更进一步的代谢转化,甲基化作用是其代谢转化的中间环节,槲皮素及甲基槲皮素的硫酸化和葡糖醛酸化是槲皮素代谢中的重要代谢通路。
Objective To investigate the metabolic characteristics of quercetin during transmembrane transportation by analyzing the metabolites of quercetin in human colon adenocarcinoma cells(Caco-2). Methods The metabolites of quercetin in Caco-2 cell monolayers were enriched by thin-layer chromatography(TLC),and their possible structures were preliminarily investigated with the LC-MS and LC-MS/MS methods. Results The metabolites of quercetin in the co-incubated Caco-2 cell lysate were analyed by full scanning mode of LC-MS,via the comparison with the blank Caco-2 cell lysate(the Caco-2 cells incubated with PBS instead of quercetin). The possible metabolites enriched by TLC were analyzed by the LC-MS and LC-MS/MS with flow injection scanning mode. The LC-MS and LC-MS/MS analysis showed that the newly detected plausible metabolites of quercetin in the Caco-2 cell monolayers included quercetin mono-sulfate,quercetin tri-sulfate,quercetin glucuronide,quercetin glucuronide sulfate,methylquercetin sulfate and methylquercetin glucuronide. No possible metabolites with m/z>600 were observed. Conclusion Quercetin underwent a broad metabolization during the transmembrane transportation simulated with the Caco-2 monolayer model,and the main metabolic reaction was methylation,sulfation and glucuronidation. The methylation of quercetin also provided an intermediate for further metabolization.The sulfation and glucuronidation of quercetin and methlquercetin are likely an important metabolic pathway in the quercetin metabolization.
作者
李素云
李峥
王立芹
高蔚娜
张振清
郭长江
LISu-yun;LI Zheng;WANG Li-qin;GAO Wei-na;ZHANG Zhen-qing;GUO Chang-jiang(Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China;Tianjin Institute of Environmental & Operational Medicine, Academy of Military Medical Sciences, Tianjing 300050, China;Institute of Pharmacology & Toxicology, Academy of Military Medical Sciences, Beijing 100850, China;Department of Epidemiology and Statistics, Hebei Medical University, Shijiazhuang 050011, China)
出处
《国际药学研究杂志》
CAS
北大核心
2019年第2期123-129,136,共8页
Journal of International Pharmaceutical Research
基金
国家自然科学基金面上项目资助(81372988)
关键词
槲皮素
Caco-2单层细胞
代谢特征
代谢产物
跨膜转运
quercetin
Caco-2 cell monolayer
metabolic characteristics
metabolite
transmembrane transport
