A novel model to assess developmental toxicity of dihaloalkanes in humans: bioactivation of 1,2-dibromoethane by the isozymes of human fetal liver glutathione S-transferase.
Glutathione S-transferase (GST) isozymes from human fetal liver (16-18 weeks gestation) were purified by affinity chromatography followed by ion-exchange high performance liquid chromatography (HPLC). The purified isozymes were used to investigate toxicity of 1,2-dibromoethane(EDB) in an in vitro model of rat embryos in culture as passive targets. At least five isozymes of GST were found in the human fetal liver. Two anionic forms [pI values 5.5 (P-2) and 4.5 (P-3)] and one basic form [pI value 8.7 (P-6)] were clearly separated. The presence of two near-neutral forms was also identified. All the isozymes of the human fetal liver GSTs tested metabolized EDB (specific activities were 2.1, 7.0, and 2.0 mumol of GSH consumed/min/mg protein for P-2, P-3, and P-6 isozymes, respectively). Covalent binding of EDB to DNA and protein was 144% and 212% higher, respectively, with the P-3 anionic isozyme when compared to the P-6 basic isozyme of GST. No covalent binding to either protein or DNA was observed with the P-2 isozyme. EDB bioactivation by the GST isozyme P-3 (15 units; 1 unit = 1 nmol of GSH consumed/min) resulted in toxicity to cultured rat embryos. Significant reductions of crown rump length, yolk sac diameter, and the composite score of morphological parameters (Brown and Fabro method) were observed. The central nervous system, optic and olfactory systems, and the hind limb were most significantly affected. The results of this investigation suggest that EDB may be classified as a suspected developmental toxicant in humans.
Mitra
A
A
Florida Toxicology Research Center, College of Public Health, University of South Florida, Tampa 33612-3899.
Hilbelink
D R
DR
Dwornik
J J
JJ
Kulkarni
A
A
D000818:Animals; D001711:Biotransformation; D004247:DNA / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism; D015946:Ethylene Dibromide / Q000378:metabolism / Q000633:toxicity*; D005260:Female; D005333:Fetus / Q000187:drug effects* / Q000378:metabolism; D005982:Glutathione Transferase / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D007527:Isoenzymes / Q000494:pharmacology*; D008099:Liver / Q000201:enzymology; D009924:Organ Culture Techniques; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity
D007527:Isoenzymes; D015946:Ethylene Dibromide; D004247:DNA; D005982:Glutathione Transferase