Browse > Article

Differential Role of Solvents on Human Cytochrome P450 2El Activity in Intact HepG2 Cells  

최달웅 (고려대학교 병설 보건대학 환경보건과)
Publication Information
Journal of Environmental Health Sciences / v.29, no.3, 2003 , pp. 9-15 More about this Journal
Abstract
The modification of CYP2El activity is a matter of considerable interest because of its role in the metabolic activation of a variety of environmental toxicants. In the present study, the time-course of changes in human CYP2El activities was determined following treatment with solvents (acetone, dimethylsulphoxide or pyridine) using intact HepG2 cells transfected by human CYP2El. Hydroxylation of chlorzoxazone was used for the measurement of CYP2El activity. CYP2E1 protein level was increased upon cultivation of cells in the presence of the solvents for 24 hr. Determination of CYP2El activities after 24 ht cultivation with the solvents demonstrated that acetone or dimethylsulphoxide increased, whereas pyridine inhibited the activities. This differential effect of the solvents on CYP2El activities persisted to subsequent 24 ht. Competitive inhibition study suggested that pyridine has stronger binding affinity to CYP2E1 than acetone or dimethylsulphoxide. These results demonstrate that different binding affinity of the solvents to CYP2El plays important role in determining real CYP2El activity in intact cells after exposure to the solvents. Present study would be helpful in precise understanding of human CYP2El-mediated toxicity.
Keywords
Cytochrome P450 2E1 (CYP2E1); Activity; Acetone; Dimethylsulphoxide; Pyridine;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Lieber, C. S. : Cytochrome P-4502El: its physiological and pathological role. Physiological Review, 77, 17-44, 1997
2 Wu, D. F., Clejan, L., Potter, B. and Cederbaum, A. I. Rapid decrease of cytochrome P-450IIE1 in primary hepatocyte culture and its maintenance by added 4-methylpyrazole. Hepatology 12, 1379-1389, 1990   DOI   PUBMED
3 Roberts, B. J., Song, B. J., Soh, Y., Park, S. S. and Shoaf, S. E. Ethanol induces CYP2El by protein stabilization. Role of ubiquitin conjugation in the rapid degradation of CYP2E1. Journal of Biological Chemistry, 270, 29632-29635, 1995   DOI
4 Yang, M. X. and Cederbaum, A. I. : Characterization of cytochrome P4502El turnover in transfected HepG2 cells expressing human CYP2E1. Archives of Biochemistry and Biophysics, 341, 25-33, 1997   DOI   ScienceOn
5 Chen, Q. and Cederbaum, A.I. : Cytotoxicity and apoptosis produced by cytochrome P450 2El in Hep G2 cells. Molecular Pharmacology, 53, 638-648, 1998   DOI
6 Chittur, S. V. and Tracy, T. S. : Rapid and sensitive high-performance liquid chromatographic assay for 6-hydrowychlorzoxazone and chlorzoxazone in liver microsomes. Journal of Chromatography b, 693, 479-484, 1997   DOI   ScienceOn
7 Carroccio, A., Wu, D. and Cederbaum， A.l. : Ethanol increases content and activity of human cytochrome P4502El in a transduced HepG2 cell line. Biochemical and Biophysical Research Communication, 203, 727-733, 1994   DOI   ScienceOn
8 Guengerich, F. P. and Turvy, C. G. Comparison of levels of several human microsomal cytochrome P- 450 enzymes and epoxide hydrolase in normal and disease states using immunochemical analysis of surgical liver samples. Journal of Pharmacology and Experimental Therapeutics, 256, 1189-1194, 1991
9 Song, B. J., Gelboin, H. V., Park, S. S., Yang, C. S. and Gonzalez, F. J. : Complementary DNA and protein sequences of ethanol-inducible rat and human cytochrome P-450s. Transcriptional and post-transcriptional regulation of the rat enzyme. Journal of Biological Chemistry 261, 16689-16697, 1986
10 Song, B. J., Veech, R. L., Park, S. S., Gelboin, H. V. and Gonzalez, F. J. Induction of rat hepatic Nnitrosodimethylamine demethylase by acetone is due to protein stabilization. Journal of Biological Chemistry, 264, 3568-3572, 1989
11 Reinke, L. A., Sexter, S. H. and Rikans, L. E. : Comparison of ethanol and imidazole pretreatments on hepatic monooxygenase activities in the rat. Research Communications in Chemical Pathology and Pharmacology, 47, 97-106, 1985
12 Kim, S. G., Williams, D. E., Schuetz, E. G. : Guzelian. P. S. and Novak, R. F. : Pyridine induction of cytochrome P-450 in the rat: role of P-450J(alcoholinducible form) in pyridine N-oxidation. Journal of Pharmacology and Experimental Therapeutics, 246, 1175-1182, 1988
13 Peter, R., Bocker, R., Beaune ， P. H., Iwasaki, M., Guengerich, F. P. and Yang, C. S. : Hydroxylation of chlorzoxazone as a specific probe for human liver cytochrome P-450IIE1. Chemical Research in Toxicology, 3, 566-573, 1990   DOI   ScienceOn
14 Patten, C. J., Ishizaki, H., Aoyama, T., Lee, M., Ning, S. M., Huang， W., Gonzalez, F. J. and Yang, C. S. : Catalytic properties of the human cytochrome P450 2E1 produced by cDNA expression in mammalian cells. Archives of Biochemistry and Biophysics, 299, 163-171. 1992   DOI   ScienceOn
15 Lowry, O, H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. : Protein measurement with folin phenol reagent. Journal of Biological Chemistry, 193, 265-275, 1951
16 Towbin. H., Staehelin, T. and Gordon, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States America, 76, 4350- 4354. 1979   DOI   ScienceOn
17 Kim, S. G. and Novak, R. F. : The induction of cytochrome P4502El by nitrogen- and sulfur-containing heterocycles: expression and molecular regulation Toxicology and Applied Pharmacology, 120, 257-265, 1993   DOI   ScienceOn
18 Eliasson, E., Johansson, I. and Ingelman-Sundberg, M. : Ligand-dependent maintenance of ethanol-inducible cytochrome P-450 in primary rat hepatocyte cell cultures. Biochemical and Biophysical Research Communication, 150, 436-443, 1988   DOI   ScienceOn
19 Yang, M. X. and Cederbaum, A. I. : Glycerol inιreases content and activity of human cytochrome P-4502E1 in a transduced HepG2 cell line by protein stabilization. Alcoholism: Clinical Experimental Research, 21, 340-347, 1997   DOI   ScienceOn
20 Ryan, D. E., Koop, D. R., Thomas, P. E., Coon, M. J. and Levin, W. : Evidence that isoniazid and ethanol induce the same microsoma1 cytochrome P-450 in rat liver, an isozyme homologous to rabbit liver cytochrome P-450 isozyme 3a. Archives of Biochemistry and Biophysics, 246, 633-644, 1986   DOI   ScienceOn
21 Hargreaves, M. B., Jones, B. J., Smith, D. A. and Gescher, A. Inhibition of p-nitrophenol hydroxylase in rat liver rnicrosomes by small aromatic and heterocyclic molecules, Drug Metabolism and Disposition, 22, 806-810, 1994