ABSTRACT
OBJECTIVES: In order to gain a better understanding of the mechanism of DMF toxicity, recent studies have focused on hepatic drug metabolizing enzymes. In this study, we investigated the effects of DMF on the induction of P450 and the activities of other related enzymes in rat liver microsomes. METHODS: DMF was administered to male Sprague Daweley rats by intraperitoneal injection at 0(control), 450(D1), 900(D2), 1,800(D3) mg DMF/kg body weight in olive oil once a day for three days. Hepatic P450 was measured by method of Omura and Sato. We evaluated selective assays for the three drug metabolizing cytochrome P450 isoenzymes 1A1, 2B1 and 2E1. RESULTS: The content of microsomal protein, P450 and b5 were tended to be decreased in DMF treated group, but they were not statistically significant. The activity of NADPH-cytochrome P450 reductase was significantly increased dose dependently(p<0.01), but the activity of NADH-b5 reductase was decreased in the treated group(p<0.01). The activities of PROD and EROD were not significant between control and treated group. The activities of pNPH in the DMF treated groups were higher than that of the control group(p<0.01). When Western immunoblottings were carried out utilizing three monoclonal antibodies which were specific against P4501A1/1, P4502B1/2 and P4502E1, the strong density band corresponding to P4502E1 was observed with the microsomes obtained from the rats treated with DMF. But there were no significant increased in the P4501A1/2 and P4502B1/2 band densities in immunoblotting. CONCLUSIONS: These result suggested that P4502E1 was inducible by DMF and P4502E1 isozyme might be responsible for the hydroxylation of DMF to HMMF.
Subject(s)
Animals , Humans , Male , Rats , Antibodies, Monoclonal , Blotting, Western , Body Weight , Cytochrome P-450 CYP1A1 , Cytochrome P-450 CYP2B1 , Cytochrome P-450 Enzyme System , Cytochromes , Dimethylformamide , Hydroxylation , Immunoblotting , Injections, Intraperitoneal , Isoenzymes , Microsomes , Microsomes, Liver , NADPH-Ferrihemoprotein Reductase , Olea , Oxidoreductases , Rats, Sprague-Dawley , Olive OilABSTRACT
The effects of styrene on the induction of cytochrome P-450s (P450), (P4501A1/2, P4502B1/2 and P4502El) and activities of other related enzymes were investigated in the male Sprague Dawley rats which were treated with styrene 500 (S1), 1,000 (S2) 1,500 (S3) mg/kg in olive oil intraperitoneally once a day for two days and sacrificed for the preparation of liver microsomes after 24 hrs. 1. The contents of total protein and P450 in the microsomes derived from the styrene treated groups were slightly higher than those from the control group except those from the 53 group (1,500 mg styrene/kg body weight) . The decreases in microsomal protein contents was prominent in the S3 (p<0.05), but the P450 contents was increased significantly in the S2 (p<0.05). 2. The activities of NADPH-P450 and NADH b5 reductase in hepatic microsomes derived from the treated groups were significantly increased in the treated groups(p<0.05). 3. The activities of PROD were also prominently increased with the treatment of styrene except in 53 group, but the activity of EROD was decreased by styrene treatment. The activities of pNPH in the styrene treated groups were higher than that of the control group (p<0.05). 5. Western blotting with monoclonal antibodies against P4502B1/2 isozymes showed the presence of P4502B1/2 in hepatic microsomes from the rats treated with styrene, and the increase in the densities of immunoblots were corelated with the dosages of styrene. The blot densities against P4501A1/2 and P4502El were slightly increased in the styrene treated groups compared with the control group. These results suggested that styrene could primarily induce P4502B1/2 as major and P4501A1/2 and P4502El in minor forms for the metabolism of styrene in rats.
Subject(s)
Animals , Humans , Male , Rats , Antibodies, Monoclonal , Blotting, Western , Cytochrome P-450 CYP1A1 , Cytochrome P-450 CYP2B1 , Cytochrome P-450 Enzyme System , Cytochromes , Isoenzymes , Metabolism , Microsomes , Microsomes, Liver , NAD , Olea , Oxidoreductases , Rats, Sprague-Dawley , StyreneABSTRACT
The effects of styrene on the induction of cytochrome P-450s (P450), (P4501A1/2, P4502B1/2 and P4502El) and activities of other related enzymes were investigated in the male Sprague Dawley rats which were treated with styrene 500 (S1), 1,000 (S2) 1,500 (S3) mg/kg in olive oil intraperitoneally once a day for two days and sacrificed for the preparation of liver microsomes after 24 hrs. 1. The contents of total protein and P450 in the microsomes derived from the styrene treated groups were slightly higher than those from the control group except those from the 53 group (1,500 mg styrene/kg body weight) . The decreases in microsomal protein contents was prominent in the S3 (p<0.05), but the P450 contents was increased significantly in the S2 (p<0.05). 2. The activities of NADPH-P450 and NADH b5 reductase in hepatic microsomes derived from the treated groups were significantly increased in the treated groups(p<0.05). 3. The activities of PROD were also prominently increased with the treatment of styrene except in 53 group, but the activity of EROD was decreased by styrene treatment. The activities of pNPH in the styrene treated groups were higher than that of the control group (p<0.05). 5. Western blotting with monoclonal antibodies against P4502B1/2 isozymes showed the presence of P4502B1/2 in hepatic microsomes from the rats treated with styrene, and the increase in the densities of immunoblots were corelated with the dosages of styrene. The blot densities against P4501A1/2 and P4502El were slightly increased in the styrene treated groups compared with the control group. These results suggested that styrene could primarily induce P4502B1/2 as major and P4501A1/2 and P4502El in minor forms for the metabolism of styrene in rats.