Formation Of Several Metabolites Research Articles

Immunochemical study on the contribution of hypolipidaemic-induced cytochrome P-452 to the metabolism of lauric acid and arachidonic acid

The influence of four hypolipidaemic drugs (clofibrate, WY-14,643, clobuzarit and bezafibrate) on hepatic cytochrome P-450 and fatty acid metabolism in male rat liver microsomes has been investigated. All of the hypolipidaemic drugs tested significantly induced the hydroxylation of lauric acid and, furthermore, this was accompanied by a concomitant 3-fold induction of a specific isoenzyme of cytochrome P-450 (termed cytochrome P-452) as determined by a single radial immunodiffusion technique. In addition, immunochemical quantitation of cytochrome P-452 in control, uninduced rat liver microsomes revealed that this particular isoenzyme constituted 22% of the total carbon monoxide-discernible cytochrome P-450 population. This has led us to the conclusion that cytochrome P-452 is a constitutive cytochrome P-450 isoenzyme and therefore that hypolipidaemic agents function as inducers of constitutive haemoprotein isoenzymes. Cytochrome P-452 plays a significant role in the hydroxylation of lauric acid as evidenced by inhibition of hydroxylase activity in the presence of an anti-P-452 IgG fraction. In addition, this antibody preferentially inhibits the 12-hydroxylation of lauric acid in rat liver microsomes by comparison to the 11-hydroxylase activity. Our studies have also shown that arachidonic acid serves as an excellent substrate for hypolipidaemic-induced cytochrome P-452, resulting in the formation of several metabolites that have been separated by reverse phase HPLC. Furthermore, a specific metabolite (or group of metabolites) of arachidonic acid is induced by clofibrate pretreatment and that the formation of this metabolite(s) is inhibited by an antibody to cytochrome P-452. By comparison, other metabolites of arachidonic acid remain refractory to induction by clofibrate and are not inhibited by the presence of anti-P-452 IgG. In addition, a reconstituted enzyme system containing highly purified cytochrome P-452 actively catalyses the above specific oxidation of arachidonic acid, a reaction that is significantly stimulated by the presence of cytochrome b 5. Taken collectively, our data provide compelling evidence that hypolipidaemic agents induce a specific isoenzyme of hepatic microsomal P-450 that readily oxidizes fatty acids and that arachidonic acid may serve as an excellent endogenous substrate for this novel haemoprotein.

Metabolism of caffeine by mouse liver microsomes: GSH or cytosol causes a shift in products from 1, 3, 7-trimethylurate to a substituted diaminouracil

Incubation of [ 14C] caffeine with hepatic microsomes from male AKR/J mice resulted in the formation of several metabolites including 1, 3, 7-trimethylurate and 6-amino-5-(N-formylmethyl-amino)-1, 3-dimethyluracil. These two compounds comprised about 60% of products and are major urinary metabolites in several animals. When cytosol was included during incubation, there was a 14-fold increase in yield of the uracil at the expense of the urate; the combination of the two metabolites remained about 60% of total products. Cytosol alone was catalytically inert. Glutathione and other sulfhydryl compounds reproduced the effect of cytosol, and the action of cytosol was accounted for quantitatively by its sulfhydryl content. We propose that an oxidized intermediate of caffeine en route to trimethylurate is reduced by glutathione to the ring-opened uracil derivative.

Dose dependent induction of rat liver microsomal cytochrome P-450 and microsomal enzymatic activities after inhalation of toluene and dichloromethane.

Sprague-Dawley rats were exposed, by inhalation, to toluene and dichloromethane (500, 1,500 or 3,000 p.p.m.) and to benzene (1,500 p.p.m.) for three days. Toluene and benzene increased the concentration of liver microsomal cytochrome P-450. A dose dependent increase in the in vitro liver microsomal formation of several metabolites of biphenyl and benzo(a)pyrene was observed for both dichloromethane and toluene. At the highest dose-level the increase in the vitro formation of benzo(a)pyrene-7,8-dihydrodiol was more than three-fold for both dichloromethane and toluene whereas the formation of benzo(a)pyrene-4,5-dihydrodiol increased more than five-fold following exposure to toluene but less than two-fold after exposure to dichloromethane. Our results suggest that dichloromethane and toluene can modify the metabolism and thereby the toxicity of other environmental contaminants.

Nature des produits de la désiodation des hormones thyroidiennes marquées simultanément par le tritrium et l'iode radioactif

1. 1. The deiodination of thyroid hormones has been studied with slices and in homogenates of rat liver, muscle and kidney, as substrates using 3,5,3′-triiodothyronine and thyroxine labeled either with tritium or with both tritium and radioactive iodine. 2. 2. The simultaneous evolution of the deiodination and degradation of the side chain of the hormones result in the formation of several metabolites. However, the formation of a tritiated and iodinated compound with an R F =o (PX) has been established as a general phenomenon in all the solvents used. At the same time small quantities of tritiated degradation products of the hormones have been identified as iodothyroacetic acid and presumably tyrosine and thyronine. 3. 3. The kinetics of the formation of the compound PX indicate that the latter is an intermediate in the deiodination of 3,5,3′-triiodothyronine and thyroxine. When isolated from a medium containing 3,5,3′-triiodo[ 3H]thyroxine, the compound PX appears to be associated with a protein. After enzymic hydrolysis, PX gives rise to tritium-labeled monoiodotyrosine, diiodotyrosine and tyrosine which suggests that their hormonal precursor might be bound to the tritium-labeled protein (PX). The formation of PX thus appears to tbe a normal step on the deiodination pathway of thyroid hormones.