Metabolic profiling of five flavonoids from Dragon's Blood in human liver microsomes using high-performance liquid chromatography coupled with high resolution mass spectrometry
Metabolic profiling of five flavonoids from Dragon's Blood in human liver microsomes using high-performance liquid chromatography coupled with high resolution mass spectrometry
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DOI：
10.1016/j.jchromb.2017.03.022
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发表时间：
2017-05-01
影响因子：
3
通讯作者：
Ren, Wei
中科院分区：
文献类型：
--
作者：
Li, Yujuan;Zhang, Yushi;Ren, Wei
虽然人们对龙血竭的药理活性了解很多，但其在人肝微粒体（HLM）和细胞色素P450（CYP）酶中的代谢尚未研究。本研究旨在确定五种黄酮类化合物（龙血素A、龙血素B、龙血素C、7，4 '-二羟基黄酮和5，7，4'-三羟基黄烷酮）在HLM中的代谢特征以及参与其代谢的代谢酶。使用高分辨率质谱法表征其代谢物的结构，并使用10种cDNA表达的CYP酶（CYP 1A 2、CYP 2A 6、CYP 2B 6、CYP 2C 8、CYP 2C 9、CYP 2C 19、CYP 2D 6、CYP 2 E1、CYP 3A 4和CYP 3A 5）验证哪些同工酶介导代谢物的代谢。根据高分辨MSn数据，共鉴定了29个代谢产物，其中龙血素A代谢产物10个，龙血素B代谢产物10个，龙血素C代谢产物4个，7，4 '-二羟基黄酮代谢产物2个，5，7，4'-三羟基黄烷酮代谢产物3个。五种黄酮类化合物在HLMs中的代谢模式包括羟基化、氧化和去甲基化。其中，羟基化是HLM中五种黄酮类化合物的主要生物转化，与其他代谢反应相结合。重组P450酶的检测结果表明，CYP 2C 9和CYP 2C 19在HLM中黄酮类化合物的羟基化过程中起重要作用。据我们所知，这是首次对HLM中龙血素A、龙血素B、龙血素C、7，4 '-二羟基黄酮和5，7，4'-三羟基黄烷酮的代谢特征进行体外评价。(C)2017爱思唯尔B. V.保留所有权利。
Although much is known about the pharmacological activities of Dragon's Blood (DB, a traditional Chinese herb), its metabolism in human liver microsomes (HLMs) and the cytochrome P450 (CYP) enzymes has not been studied. This study aims to identify the metabolic profile of five flavonoids (loureirin A, loureirin B, loureirin C, 7,4'-dihydroxyflavone and 5,7,4'-trihydroxyflavanone) from DB in HLMs as well as the CYP enzymes that are involved in the metabolism of them. High-resolution mass spectrometry was used to characterize the structures of their metabolites and 10 cDNA-expressed CYP enzymes (CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4 and CYP3A5) were used to verify which isozymes mediate in the metabolism of the metabolites. Totally, 29 metabolites including 10 metabolites of loureirin A, 10 metabolites of loureirin B, 4 metabolites of loureirin C, 2 metabolites of 7,4'-dihydroxyflavone and 3 metabolites of 5,7,4'-trihydroxyflavanone were elucidated and identified on the basis of the high-resolution MSn data. The metabolic profile of the five flavonoids in HLMs involved hydroxylation, oxidation and demethylation. Among them, hydroxylation was the predominant biotransformation of the five flavonoids in HLMs, occurring in combination with other metabolic reactions. Assay with recombinant P450s revealed that CYP2C9 and CYP2C19 played an important role in the hydroxylation of flavonoids in HLMs. To the best of our knowledge, this is the first in vitro evaluation of the metabolic profile of loureirin A, loureirin B, loureirin C, 7,4'-dihydroxyflavone and 5,7,4'-trihydroxyflavanone in HLMs. (C) 2017 Elsevier B.V. All rights reserved.