Preparation and analysis of a lung microsomal fraction from control and 3-methylcholanthrene treated rats.
In order to facilitate the homogenization of lung tissue it was previously incubated with collagenase during 30 minutes. Morphological observations were performed in order to ascertain the cell integrity. The enzymatically digested tissue was homogenized in a 0.25 M sucrose solution containing 1 mM EDTA, 3 mM imidazole (pH.7.3) and supplemented with 1 mM imipramine in order to stabilize the mitochondria, which otherwise might contaminate the microsomal fraction. The homogenate was then centrifuged and subdivided into four fractions which were analyzed for their content in protein and for the activities of so-called marker enzymes. The cytochrome P450 level was measured in both control and 3-methylcholanthrene preparations. The activities and the kinetic parameters of lung benzpyrene hydroxylase and aldrin epoxidase were measured using the lung microsomal fractions from control and previously 3-methylcholanthrene treated rats; 3-methylcholanthrene pretreatment modified the catalytiac properties of both enzymes.
D000818:Animals; D001579:Benzopyrene Hydroxylase / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D006868:Hydrolysis; D007099:Imipramine / Q000494:pharmacology; D008168:Lung / Q000032:analysis / Q000187:drug effects / Q000648:ultrastructure*; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology*; D003012:Microbial Collagenase; D008861:Microsomes / Q000032:analysis* / Q000187:drug effects; D006899:Mixed Function Oxygenases / Q000378:metabolism; D051381:Rats
D008748:Methylcholanthrene; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C019647:aldrin epoxidase; D001579:Benzopyrene Hydroxylase; D003012:Microbial Collagenase; D007099:Imipramine