高效液相色谱检测天麻素在不同种属肝微粒体的体外代谢差异  

Detection of in Vitro Metabolic Differences of Gastrodin in Liver Microsomes of Different Species by High Performance Liquid Chromatography

作　　者：闫杰 YAN Jie(The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000 China)

机构地区：[1]锦州医科大学附属第一医院,辽宁锦州121000

出　　处：《锦州医科大学学报》2024年第5期34-38,共5页Journal of Jinzhou Medical University

摘　　要：目的建立天麻素在肝微粒体样品中的高效液相色谱(high performance liquid chromatography,HPLC)检测方法,鉴定天麻素在不同种属的体外代谢差异。方法采用乙腈沉淀法处理天麻素肝微粒体样品,C18色谱柱(4.6 mm×250 mm,5μm)分离,建立HPLC天麻素检测方法;天麻素和不同种属肝微粒分别在Ⅰ相(cytochrome P450,CYP)和Ⅱ相(uroidine diphosphate glucuronyl transferase,UGT)孵育体系下一起孵育,应用所建立的HPLC方法检测孵育后天麻素含量变化,确定其在不同种属的代谢差异。采用分子对接方式,预测天麻素和CYP酶的相互作用情况及作用位点。结果所建立天麻素肝微粒体HPLC检测方法的精密度、准确度和稳定性均符合生物样品分析要求。天麻素在不同种属之间的代谢有明显差异,人和猴的代谢特点较为相似。CYP3A4和CYP2C19可能是天麻素的主要代谢酶,天麻素和CYP之间通过氢键相互作用。结论天麻素在不同种属的代谢差异,将为其在药理和毒理研究中实验动物的选择提供依据。Objective To establish a high-performance liquid chromatography(HPLC)detection method for gastrodin in liver microsomes and identify its in vitro metabolic differences in different species.Methods Gastrodin liver microsome samples were treated by acetonitrile precipitation and separated by C18 chromatography column(4.6 mm×250 mm,5μm)to establish HPLC method for gastrodin detection;gastrodin was incubated with different liver microsomes in phase Ⅰ(CYP)and phase Ⅱ(UGT)systems.The established HPLC method was used to detect the change of gastrodin content after incubation,and to determine the difference of its metabolism in different species.Molecular docking was applied to predict the interaction and sites between gastrodin and CYP450 enzymes.Results The precision,accuracy and stability of HPLC method for gastrodin liver microsomes met the requirements of biological sample analysis.There were significant differences in the metabolism of gastrodin among different species,and the metabolic characteristics of humans and monkeys were relatively similar.CYP3A4 and CYP2C19 may be the main metabolic enzymes of gastrodin,and gastrodin interacts with CYP through hydrogen bonding.Conclusion The metabolic differences of gastrodin among different species will provide a basis for the selection of experimental animals in future pharmacological and toxicological studies.

关 键 词：天麻素 高效液相色谱检测法 种属差异 分子对接 

分 类 号：R285[医药卫生—中药学]