Epoxide hydrolase activity in human blood mononuclear leukocytes: individual differences in native and mitogen-stimulated cells

被引:2
作者
Bernardini, S [1 ]
Norppa, H [1 ]
Elovaara, E [1 ]
机构
[1] Finnish Inst Occupat Hlth, Dept Ind Hyg & Toxicol, FIN-00250 Helsinki, Finland
关键词
drug-metabolizing enzyme; EC 3.3.2.3.(epoxide hydrolase); 1,2-epoxy-3-(p-nitrophenoxy)propane; CAS No. 5255-75-4; human lymphocytes; interindividual variation;
D O I
10.1016/S1383-5718(99)00108-4
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Epoxide hydrolase (EH; EC 3.3.2.3) activity was measured in whole-cell sonicates of native and cultured peripheral blood mononuclear cells (PBMCs) from 19 healthy unrelated Caucasian donors (age, 28-55 years). We used 1,2-epoxy-3-(p-nitrophenoxy)propane (0.34 mM) as the substrate and, for the diol assay, a quantitative HPLC method with spectrophotometric detection. One portion of the PBMCs was frozen immediately, while the other portion was PHA-stimulated and cultivated for 36 h. In native leukocytes, the EH activity varied from 2.2 to 8.2 pmol/min per 10(6) cells (3.8-fold), the mean +/- SD was 5.6 +/- 1.4 pmol/min per 10(6) cells. In most of the samples from different donors, the specific activity increased in cultivation, varying from 2.4 to 15.4 pmol/min per 10(6) cells (6.3-fold), the mean +/- SD being 8.5 +/- 3.8 pmol/min per 10(6) cells. From a methodological point of view, enzyme measurement in native cells is simple to perform and may provide a better index of the specific activity, as the accuracy of electronic cell counting is better for cell samples taken before than after cultivation. The differences in the EH activity of PBMCs indicate that significant interindividual variation may occur in the detoxification of epoxides produced in the human lymphocyte test systems commonly used for genotoxicity screening of chemicals in vitro. Further studies are needed to determine the extent to which the reproducibility and thus also the sensitivity of such assays could be improved by analyses carried out to control the donors for their EH phenotype. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:387 / 392
页数:6
相关论文
共 15 条
[1]   Induction of sister chromatid exchange by 1,2-epoxy-3-butene in cultured human lymphocytes:: influence of GSTT1 genotype [J].
Bernardini, S ;
Hirvonen, A ;
Pelin, K ;
Norppa, H .
CARCINOGENESIS, 1998, 19 (02) :377-380
[2]   SIMPLIFIED LIQUID-CHROMATOGRAPHIC ASSAY FOR EPOXIDE HYDROLASE [J].
GIULIANO, KA ;
LAU, EP ;
FALL, RR .
JOURNAL OF CHROMATOGRAPHY, 1980, 202 (03) :447-452
[3]  
GLATT H, 1980, CANCER RES, V40, P2552
[4]  
Glatt HR, 1984, BIOCH BASIS CHEM CAR, P107
[5]   THE HUMAN MICROSOMAL EPOXIDE HYDROLASE GENE (EPHX1) - COMPLETE NUCLEOTIDE-SEQUENCE AND STRUCTURAL CHARACTERIZATION [J].
HASSETT, C ;
ROBINSON, KB ;
BECK, NB ;
OMIECINSKI, CJ .
GENOMICS, 1994, 23 (02) :433-442
[6]   Human hepatic microsomal epoxide hydrolase: Comparative analysis of polymorphic expression [J].
Hassett, C ;
Lin, J ;
Carty, CL ;
Laurenzana, EM ;
Omiecinski, CJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1997, 337 (02) :275-283
[7]   INFLUENCE OF THE LEVEL OF CYTOSOLIC EPOXIDE HYDROLASE ON THE INDUCTION OF SISTER CHROMATID EXCHANGES BY TRANS-BETA-ETHYLSTYRENE 7,8-OXIDE IN HUMAN-LYMPHOCYTES [J].
KRAMER, A ;
FRANK, H ;
SETIABUDI, F ;
OESCH, F ;
GLATT, H .
BIOCHEMICAL PHARMACOLOGY, 1991, 42 (11) :2147-2152
[8]   CYTOSOLIC EPOXIDE HYDROLASE [J].
MEIJER, J ;
DEPIERRE, JW .
CHEMICO-BIOLOGICAL INTERACTIONS, 1988, 64 (03) :207-249
[9]   INTERINDIVIDUAL VARIATIONS IN THE ACTIVITIES OF CYTOSOLIC AND MICROSOMAL EPOXIDE HYDROLASE IN HUMAN-LIVER [J].
MERTES, I ;
FLEISCHMANN, R ;
GLATT, HR ;
OESCH, F .
CARCINOGENESIS, 1985, 6 (02) :219-223
[10]   ROLE OF GSTT1 AND GSTM1 GENOTYPES IN DETERMINING INDIVIDUAL SENSITIVITY TO SISTER-CHROMATID EXCHANGE INDUCTION BY DIEPOXYBUTANE IN CULTURED HUMAN-LYMPHOCYTES [J].
NORPPA, H ;
HIRVONEN, A ;
JARVENTAUS, H ;
UUSKULA, M ;
TASA, G ;
OJAJARVI, A ;
SORSA, M .
CARCINOGENESIS, 1995, 16 (06) :1261-1264