Inhibition of gonadotropin action and progesterone synthesis by xanthine oxidase in rat luteal cells.
Hydrogen peroxide produces marked antigonadotropic and lytic actions in luteal cells, but the effects of superoxide, the archetypal oxygen radical, are unknown. Xanthine oxidase generates superoxide, and the activity of this enzyme, and purine substrate, are increased under ischemia, such as that seen at luteal regression. We therefore examined the actions of xanthine oxidase on luteal cells to assess the effects of this enzyme and the superoxide anion on luteal function. Xanthine oxidase, in the presence of hypoxanthine (50 microM), produced marked inhibition of LH-sensitive cAMP and progesterone production with complete inhibition at 25 mU/ml and half-maximal inhibition at about 5 mU/ml. These antigonadotropic actions of xanthine oxidase were rapid with maximal effects within 5 min, followed several minutes later by substantial depletion of ATP. Heat, superoxide dismutase, and catalase or catalase alone abolished the actions of xanthine oxidase. While depletion of ATP by xanthine oxidase was prevented by 3-amino-benzamide, an inhibitor of DNA repair, inhibition of cAMP and progesterone production was still evident. Xanthine oxidase also inhibited progesterone synthesis stimulated by 8-bromo-cAMP. Isobutylmethylxanthine, a cAMP phosphodiesterase inhibitor, did not reverse the inhibition of cAMP accumulation by xanthine oxidase, and the enzyme had no effect on LH receptor binding activity. Since catalase reversed the effects of xanthine oxidase, we conclude that superoxide was rapidly dismuted to hydrogen peroxide and mediated the antigonadotropic and antisteroidogenic actions of xanthine oxidase in luteal cells. The sensitivity of luteal cells to xanthine oxidase raises the possibility that this enzyme may serve as a significant source of hydrogen peroxide in the corpus luteum.
Gatzuli
E
E
Department of Obstetrics/Gynecology, Yale University School of Medicine, New Haven, Connecticut 06510.
Aten
R F
RF
Behrman
H R
HR
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D006063:Chorionic Gonadotropin / Q000378:metabolism; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D000242:Cyclic AMP / Q000502:physiology; D005260:Female; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011374:Progesterone / Q000096:biosynthesis*; D051381:Rats; D011974:Receptors, LH / Q000378:metabolism; D014969:Xanthine Oxidase / Q000494:pharmacology*
D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D015124:8-Bromo Cyclic Adenosine Monophosphate; D011374:Progesterone; D000255:Adenosine Triphosphate; D007986:Luteinizing Hormone; D006861:Hydrogen Peroxide; D000242:Cyclic AMP; D014969:Xanthine Oxidase; D015056:1-Methyl-3-isobutylxanthine