模式生物斑马鱼用于5-羟基黄酮代谢的合理性研究

Metabolism of 5-Hydroxyflavone in Model Organism Zebrafish

关键词:  斑马鱼  5-羟基黄酮  Ⅱ相代谢  高效液相色谱-电喷雾质谱

Key words:  zebrafish  5-hydroxyflavone  phase Ⅱ metabolism  HPLC-ESI/MS

摘要:

目的 采用模式生物斑马鱼研究5-羟基黄酮的代谢，探索斑马鱼用于药物Ⅱ相代谢的适用性。方法 将斑马鱼培养于5-羟基黄酮溶液中，定时取鱼体及药液，采用高效液相色谱-电喷雾质谱联用检测，根据正、负离子模式准分子离子峰获得化合物分子量信息，通过与文献数据或对照品对照，结合碎片离子，推测可能的代谢产物。结果 在斑马鱼体内或体外药液检测到5-羟基黄酮原型及其2个单羟基葡萄糖醛酸结合物和1个单羟基硫酸结合物。结论 5-羟基黄酮在斑马鱼作用下的葡萄糖醛酸化反应与其在大鼠体内的Ⅱ相代谢机制高度一致，并首次在负离子模式下检测到5-羟基黄酮硫酸结合物。斑马鱼用于药物Ⅱ相代谢具合理性，且具有化合物用量少、成本低、方法简单、高效的优势，为建立斑马鱼体内药物代谢新模型提供重要参考。

Abstract:

OBJECTIVE Model organism zebrafish was used to study the metabolism of 5-hydroxyflavone to investigate the feasibility of zebrafish in drug phase Ⅱ metabolism. METHODS Zebrafish was exposed to 5-hydroxyflavone solution for 24 h, and the samples of the solution and the zebrafish bodies were analyzed by high performance liquid chromatography- electrospray mass spectrometry (HPLC-ESI-MS) method. The quasi-molecular ions of the compounds in both negative and positive modes were observed for molecular mass information, and the potential structures were identified by attentive studying on the mass spectra of the compounds and comparing with reference data or standards. RESULTS In addition to parent compound, 3 phase Ⅱ metabolites were identified，including two glucuronidation products and one sulfation product. CONCLUSION Glucuronidation of 5-hydroxyflavone in zebrafish was highly consistent with that in rat, in addition, 5-hydroxyflavone sulfate was detected for the first time in negative mode. The present results indicate that it is reasonable to apply zebrafish in imitating current phase Ⅱ metabolism. Zebrafish model has the advantages of far less compound-consuming, lower cost, simpler procedure and higher efficiency, which provide important reference for making it a novel in vivo model of drug metabolism.