Induction of antioxidant enzyme activities by a phenylurea derivative, EDU.
Oxygen free radicals have the potential to mediate cell injury. Defenses against such radicals include the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX). The purposes of this study were (1) to develop an in vitro model using human cells in which to investigate a potential pharmacologic agent as an inducer of these antioxidant enzymes; (2) to investigate the phenylurea derivative N-[2-(2-oxo-1-imidazolindinyl)ethyl]-N-phenylurea (EDU) in this model with paraquat (PQ) serving as the positive control; and (3) to determine if induction of the antioxidant enzymes by EDU occurs in vivo. Human gingival fibroblasts (Gin-1) were used as the target cell in vitro; PQ and EDU, an inducer of SOD and CAT activities in plants, were evaluated as antioxidant enzyme inducers. Total SOD activity in Gin-1 cells increased 2-fold (p less than 0.05) in the presence of 1.0 mM PQ for 18-48 hr compared with untreated controls. Gin-1 cells incubated with 0.25-2.0 mM PQ for 24 hr had significantly increased total SOD (1.5 to 2.0-fold; p less than 0.05). CAT activity increased with 1.0 and 2.0 mM PQ (p less than 0.05). In the presence of PQ, GSH-PX activity decreased (p less than 0.05) in a concentration-dependent manner, indicating inactivation of this enzyme. No toxicity, indicated by lactate dehydrogenase released into the incubation medium, was noted at PQ concentrations below 5.0 mM. In the presence of 0.125-2.0 mM EDU, total SOD activity in Gin-1 cells significantly increased (1.5 to 2.0-fold; p less than 0.05). CAT activity significantly increased in a dose-dependent manner (p less than 0.05), while GSH-PX activity remained constant following exposure to 0.125-2.0 mM EDU. Intraperitoneal administration of EDU to rats twice a day for 2 days at 100 mg/kg induced SOD activity in heart, liver, and lung compared to controls (p less than 0.05). CAT activity increased in the liver 56% and in the lung 36% (p less than 0.05). GSH-PX activity remained constant. Our findings indicate that Gin-1 cells are a useful model in which to study inducers of antioxidant enzymes in vitro and that the phenylurea compound EDU induces SOD and CAT activities both in vitro and in vivo.
Stevens
T M
TM
Medical Products Department, E. I. du Pont de Nemours & Company, Inc., Wilmington, Delaware 19898.
Boswell
G A
GA
Adler
R
R
Ackerman
N R
NR
Kerr
J S
JS
D000975:Antioxidants / Q000378:metabolism*; D002374:Catalase / Q000096:biosynthesis; D002460:Cell Line; D002701:Chloramphenicol / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004790:Enzyme Induction; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology; D005609:Free Radicals; D005979:Glutathione Peroxidase / Q000096:biosynthesis; D006801:Humans; D010269:Paraquat / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000096:biosynthesis
D000975:Antioxidants; D005609:Free Radicals; D010671:Phenylurea Compounds; D003609:Dactinomycin; C032766:ethylene diurea; D002701:Chloramphenicol; D003513:Cycloheximide; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D010269:Paraquat