Pu Xiao, Gao Yiqiao, Li Ruiting, Li Wei, Tian Yuan, Zhang Zunjian, Xu Fengguo
Key Laboratory of Drug Quality Control and Pharmacovigilance (Ministry of Education), State Key Laboratory of Natural Medicine, China Pharmaceutical University, Nanjing 210009, China.
Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing 210009, China.
Metabolites. 2019 Apr 18;9(4):77. doi: 10.3390/metabo9040077.
Cytochrome P450 1A2 (CYP1A2) is one of the major CYP450 enzymes (CYPs) in the liver, and participates in the biotransformation of various xenobiotics and endogenous signaling molecules. The expression and activity of CYP1A2 show large individual differences, due to genetic and environmental factors. In order to discover non-invasive serum biomarkers associated with hepatic CYP1A2, mass spectrometry-based, untargeted metabolomics were first conducted, in order to dissect the metabolic differences in the serum and liver between control rats and β-naphthoflavone (an inducer of CYP1A2)-treated rats. Real-time reverse transcription polymerase chain reaction and pharmacokinetic analysis of phenacetin and paracetamol were performed, in order to determine the changes of mRNA levels and activity of CYP1A2 in these two groups, respectively. Branched-chain amino acids phenylalanine and tyrosine were ultimately focalized, as they were detected in both the serum and liver with the same trends. These findings were further confirmed by absolute quantification via a liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based targeted metabolomics approach. Furthermore, the ratio of phenylalanine to tyrosine concentration was also found to be highly correlated with CYP1A2 activity and gene expression. This study demonstrates that metabolomics can be a potentially useful tool for biomarker discovery associated with CYPs. Our findings contribute to explaining interindividual variations in CYP1A2-mediated drug metabolism.
细胞色素P450 1A2（CYP1A2）是肝脏中主要的细胞色素P450酶（CYPs）之一，参与多种外源性物质和内源性信号分子的生物转化。由于遗传和环境因素，CYP1A2的表达和活性存在很大的个体差异。为了发现与肝脏CYP1A2相关的非侵入性血清生物标志物，首先进行了基于质谱的非靶向代谢组学研究，以剖析对照组大鼠和β-萘黄酮（CYP1A2诱导剂）处理组大鼠血清和肝脏中的代谢差异。分别进行了对乙酰氨基酚和非那西丁的实时逆转录聚合酶链反应及药代动力学分析，以确定这两组中CYP1A2的mRNA水平和活性变化。支链氨基酸苯丙氨酸和酪氨酸最终成为重点研究对象，因为它们在血清和肝脏中均被检测到且呈现相同趋势。通过基于液相色谱-串联质谱（LC-MS/MS）的靶向代谢组学方法进行绝对定量，进一步证实了这些发现。此外，还发现苯丙氨酸与酪氨酸浓度的比值与CYP1A2活性和基因表达高度相关。本研究表明，代谢组学可能是发现与CYPs相关生物标志物的潜在有用工具。我们的发现有助于解释CYP1A2介导的药物代谢中的个体差异。
