The in vitro effects of selected environmental toxicants on two heme synthesis enzymes.
Benzene and some of its substitution products become environmental toxicants due to improper disposal procedures. Benzene has been found to alter heme and globin synthesis in anucleate rabbit reticulocytes (Forte et al., 1976; Wildman et al., 1976) and based on these findings we felt it would be useful to determine what, if any, effect these derivatives would have on heme synthesis in vitro by studying their influence on delta-aminolevulinic acid synthetase (ALAS) and ferrochelatase (FC) activities in rat liver homogenates. ALAS was measured according to Ebert et al. (1970). FC was measured after Williams et al. (1980). Final concentrations of each added compound to the reaction mixture were 10(-3) to 10(-6) M. Normal values for rat liver ALAS were 250-350 nmol ALA/g protein/30 min, mean 290 +/- 40, and for FC were 12-40 mumol heme/g protein/45 min, mean 20 +/- 7. At 10(-3) M and lower concentrations these compounds inhibited ALAS and stimulated FC activities. Their effect on ALAS activity expressed as percentage of control of three analyses performed in triplicate +/- SEM was: o- and p-dinitrobenzenes-46 +/- 2; trinitrotoluenes-55 +/- 2; dinitrotoluenes-70 +/- 2; and amino-dinitrotoluenes-171 +/- 4. The stimulatory effect of these compounds expressed as percentage of control +/- SEM on FC was: dinitrotoluenes-171 +/- 3; dinitrobenzenes-152 +/- 3; trinitrotoluenes-142 +/- 4; and amino-dinitrotoluenes-130 +/- 4. Other classes of compounds tested did not significantly affect these enzymes at the same concentrations. These in vitro techniques may prove useful for predicting in vivo toxicologic effects of pollutants on species of interest.
D000624:5-Aminolevulinate Synthetase / Q000037:antagonists & inhibitors*; D000818:Animals; D002384:Catalysis; D004785:Environmental Pollutants / Q000633:toxicity*; D004789:Enzyme Activation / Q000187:drug effects; D005294:Ferrochelatase / Q000378:metabolism*; D006418:Heme / Q000096:biosynthesis*; D066298:In Vitro Techniques; D007220:Industrial Waste / Q000633:toxicity*; D008099:Liver / Q000201:enzymology; D008190:Lyases / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012997:Solvents / Q000633:toxicity
D004785:Environmental Pollutants; D007220:Industrial Waste; D012997:Solvents; D006418:Heme; D000624:5-Aminolevulinate Synthetase; D008190:Lyases; D005294:Ferrochelatase