An In vitro study on methotrexate hydroxylation in rat and human liver

Title:

An In vitro study on methotrexate hydroxylation in rat and human liver

Creator:

Chládek, J., Martínková, J., and Šišpera, L.

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:e18ad0bb-b3bb-4cd0-b183-80ce1d78acdb
uuid:e18ad0bb-b3bb-4cd0-b183-80ce1d78acdb

Subject:

methotrexate, cytochrome P450, 7-hydroxymethotrexate, menadion, allopurinol, in vitro inhibition, and rat and human liver

Type:

article, model:article, and TEXT

Description:

Methotrexate (MTX) was investigated for possible effect on the metabolism of ethoxyresorufin, pentoxyresorufin and ethjxycoumarin, the model substrates of cytochrome P450. The investigation was carried out in liver microsomes of rats pretreated with classical inducers of cytochrome P450 as well as in microsomes of two human livers. Firthermore, we measured the conversion of MTX (100 ^M) to its main metabolite, 7-hydroxymethotrexate (7-OHMTX), in microsomes and cytosolic fractions of rat and human livers. The inhibition of 7-OHMTX formation by menadion (inhibitor of aldehyde oxidase) and allopurinol (inhibitor of xanthine oxidase) was studied in the cytosol of rat and human livers. In both species, MTX in the concentration range 0.5-500 /¿M exerted no inhibitory effect on enzymatic activities associated with cytochrome P450. Moreover, we did not observe any measurable formation of 7-OHMTX in liver microsomes. MTX was metabolized at a similar rate in the cytosol of rat and human liver. Allopurinol (100 /iM) reduced the rate of MTX hydroxylation by 31.5 % in the cytosol of human livers but had no effect in the rat. Menadion (100 y/M) decreased the rate of 7-OHMTX formation in the cytosol of human and rat liver by 69 % and 94 %, respectively. Our results confirmed that MTX is oxidized by a soluble enzymatic system in both the rat and human liver. In human tissues, both aldehyde oxidase and xanthine oxidase may play an important role in the metabolism of MTX. Depression of cytochrome P450 and related enzymatic activities observed in vivo cannot be explained by a direct inhibitory action of MTX on cytochrome P450

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Source:

Physiological research | 1997 Volume:46 | Number:5

Harvested from:

CDK

Metadata only:

false