Drug-metabolizing enzymes in rat, mouse, pig and human macrophages and the effect of phagocytic activation.
Aryl hydrocarbon hydroxylase (AHH) activity mediated by cytochrome P-450 is present in pig hepatic microsomes [10 nmol.3 mg protein-1.hr-1]. AHH activity was detectable in both hepatocytes and Kupffer cells isolated from pig liver biopsy material. These cells were isolated from needle or wedge biopsy material by collagenase perfusion and incubation with collagenase at 37 degrees. The two cell types were separated from the resulting cell suspension as previously described for whole liver. Kupffer cells were enriched by adherence and were cultured for 24 hr prior to harvesting. Cells were harvested, and cell viability was determined. AHH activity was assayed in Kupffer cell and hepatocyte homogenates. Kupffer cell AHH activity was approximately one-eighth the level detected in hepatocytes. To determine whether this enzyme was present in other macrophages, monocytes were isolated from 10 ml of heparinized peripheral blood using Ficoll-Hypaque and were enriched by adherence. After 24 hr in culture, cell viability was assessed and monocytes were identified by by cytochemical staining. AHH activity was detectable in pig monocyte homogenates, and the AHH level was similar to that in pig Kupffer cells. AHH was also easily detectable in human monocytes. This macrophage AHH activity was compared with AHH activity in rat monocytes, mouse Kupffer cells and mouse peritoneal macrophages. Monocyte AHH was relatively stable in cell culture but decreased rapidly upon storage at -70 degrees. Macrophage AHH activity was depressed following phagocytic activation in vitro by latex beads with a concomitant increase in heme oxygenase activity.
Department of Medicine, Dalhousie University, Halifax, Nova Scotia, Canada.
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Peterson
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002469:Cell Separation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005260:Female; D006419:Heme Oxygenase (Decyclizing) / Q000378:metabolism; D006801:Humans; D008658:Inactivation, Metabolic; D007728:Kupffer Cells / Q000201:enzymology; D008262:Macrophage Activation; D008264:Macrophages / Q000201:enzymology* / Q000276:immunology; D008297:Male; D051379:Mice; D051381:Rats; D011919:Rats, Inbred Strains; D013045:Species Specificity; D013552:Swine
D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D006419:Heme Oxygenase (Decyclizing)