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同时测定猪肝微粒体中细胞色素P450 1A、2A和3A的活性。

Simultaneous determination of cytochrome P450 1A, 2A and 3A activities in porcine liver microsomes.

作者信息

Johansson Monika, Tomankova Jana, Li Shengjie, Zamaratskaia Galia

机构信息

Department of Food Science, BioCenter, Swedish University of Agricultural Sciences, Uppsala, Sweden.

出版信息

Interdiscip Toxicol. 2012 Sep;5(3):150-4. doi: 10.2478/v10102-012-0024-3.

DOI:10.2478/v10102-012-0024-3
PMID:23554555
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3600515/
Abstract

The aim of this study was to develop a robust method for the simultaneous determination of the activities of three porcine CYP450 enzymes in hepatic microsomes. A cocktail consisting of three selective CYP450 probe substrates, 7-ethoxyresorufin (CYP1A), coumarin (CYP2A) and 7-benzyloxy-4-trifluoromethylcoumarin (BFC; CYP3A), was incubated with porcine liver microsomes. The presence of 7-ethoxyresorufin appears to significantly influence the kinetics of coumarin hydroxylation and BFC O-debenzylation. These results indicate that the use of 7-ethoxyresorufin in substrate cocktails together with coumarin and BFC should be avoided.

摘要

本研究的目的是开发一种可靠的方法，用于同时测定肝微粒体中三种猪CYP450酶的活性。将由三种选择性CYP450探针底物7-乙氧基试卤灵（CYP1A）、香豆素（CYP2A）和7-苄氧基-4-三氟甲基香豆素（BFC；CYP3A）组成的混合物与猪肝微粒体一起孵育。7-乙氧基试卤灵的存在似乎会显著影响香豆素羟基化和BFC O-脱苄基反应的动力学。这些结果表明，应避免在底物混合物中同时使用7-乙氧基试卤灵与香豆素和BFC。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/10b0e61d4db6/ITX-5-150-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/8898d7d59fd2/ITX-5-150-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/cc5729e4b638/ITX-5-150-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/10b0e61d4db6/ITX-5-150-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/8898d7d59fd2/ITX-5-150-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/cc5729e4b638/ITX-5-150-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ce5/3600515/10b0e61d4db6/ITX-5-150-g003.jpg

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Sensors (Basel). 2009;9(3):2134-47. doi: 10.3390/s90302134. Epub 2009 Mar 23.
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An in vitro, high throughput, seven CYP cocktail inhibition assay for the evaluation of new chemical entities using LC-MS/MS.一种用于评估新化学实体的体外高通量七重细胞色素P450鸡尾酒抑制试验，采用液相色谱-串联质谱法。
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