大豆苷元在人肝微粒体中的单羟化代谢的机制论文.pdfVIP

大豆苷元在人肝微粒体中的单羟化代谢的机制 1* 1 2 彭文兴 李焕德 周宏灏 1中南大学湘雅二医院临床药学研究室 2中南大学临床药理研究所 湖南 长沙 410011 摘要 目的 探讨大豆苷元在人肝微粒中羟基化代谢所涉及的肝细胞色素P450CYP同工酶 为 研究其在人体内的代谢提供基础 方法 通过分析大豆苷元在肝微粒体中和重组 CYP酶中形成的单羟 化代谢物的酶促动力学,分析其酶学模型 然后用不同CYP同工酶选择性抑制剂或底物进行抑制实验 初步筛选出介导大豆苷元单羟化代谢所涉及的 CYP同工酶的 结果 代谢物的形成动力学符合米氏方 程单酶模型 CYP1A2选择性抑制剂呋喃茶碱和CYP1A2单克隆抗体均能明显抑制三种单羟化代谢物的 形成 而其他CYP选择性的抑制剂对三种代谢物的形成没有或较小产生抑制作用 用重组酶实验得出 相同结果 结论 体外肝微粒体研究表明 大豆苷元的单羟基代谢主要由CYP1A2所介导 关键词 大豆苷元 羟基化代谢 CYP1A2 Mono-hydroxylation mechanism of daidzein in human liver microsome PENG Wen-xing1 LI Huan-de1 ZHOU Hong-hao2 1 ( Department of Clinical Pharmacy, Second Xiang Ya Hospital, Central South University, Changsha 410011; Clinical Pharmacology Institute, Central South University, Changsha, 410078, China) Abstract: Aim In vitro studies with rats or human liver microsomes demonstrated that daidzein was readily metabolized to mono-hydroxylated compounds. In this study, daidzein mono -hydroxylated metabolites was further investigated using human liver microsomes to identify the cytochrome P450 (CYP) isoform (s) involved in this metabolic pathway. Methods Kinetic analysis of the formation rates of mono-hydroxylated metabolites of daidzein, including 7,8,4-trihydroxyisoflavone (7,8,4-THI), 7,3,4-trihydroxyisoflavone (7,3,4-THI) and 6,7,4-trihydroxyisoflavone (6,7,4-THI), were performed using human liver microsomes (HLM) and recombinant enzymes at substrate concentrations ranging from 0.5 to - 1 400 mol· L . Nine selective inhibitors or substrate probes specific for different CYP isoforms were applied for screening the isoform (s) responsible for mono-hydroxylated metabolism of daidzein. Results Michaelis-Menten kinetic par