Lipid hydroperoxides evoke antigonadotropic and antisteroidogenic activity in rat luteal cells.
At functional luteolysis, the rat corpus luteum generates hydrogen peroxide (H2O2), which is known to rapidly inhibit gonadotropin-sensitive cAMP and progesterone production in isolated luteal cells. Lipid peroxides also increase markedly in the rat corpus luteum with the onset of functional luteolysis, and H2O2 is a potent inducer of lipid peroxidation. However, the actions of lipid peroxides on cell function are unknown. The objective of this study was to investigate the impact of typical lipid peroxides, cumene hydroperoxide (CuOOH) and 15(S)-hydroperoxyeicosatetraenoic acid, on rat luteal cells. CuOOH inhibited both LH-sensitive cAMP accumulation (ED50, 25 microM) and progesterone production (ED50, 20 microM). 15(S)-hydroperoxyeicosatetraenoic acid also dose dependently inhibited steroidogenesis. A significant reduction of LH-stimulated progesterone production was evident within 5 min of treatment with CuOOH, whereas inhibition of cAMP accumulation was not evident until 60 min. 8-Bromo-cAMP and 22-hydroxycholesterol caused partial and complete reversal of CuOOH-inhibited progesterone secretion, respectively. Preincubation of cells with o-phenanthroline completely reversed the inhibitory effects of CuOOH on cAMP accumulation and partially reversed its effects on progesterone production. Incorporation of radiolabeled amino acids into luteal proteins was significantly inhibited by CuOOH (25 microM) within 2 min of treatment and was reduced to 40 +/- 14% of control levels at 60 min. CuOOH (25 microM) maximally stimulated PGE2 production within 30 min of treatment (180 +/- 30% of control), a response that was completely blocked by aristolochic acid (100 microM), a phospholipase-A2 inhibitor, and indomethacin (1 microgram/ml), a prostaglandin (PG) synthesis inhibitor. The present results suggest that the acute inhibitory action of lipid peroxides on LH-stimulated progesterone production occurs down-stream of cAMP synthesis and appears to be due to impaired cholesterol utilization for steroidogenesis, possibly through inhibition of protein synthesis. The stimulation of PGE2 production by CuOOH appears to involve the activation of phospholipase-A2, which is a rate-limiting step in PG synthesis. Lipid peroxides as well as H2O2 may serve as mediators of functional luteolysis.
Kodaman
P H
PH
Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, Connecticut 06520-8063.
Aten
R F
RF
Behrman
H R
HR
D000818:Animals; D001555:Benzene Derivatives / Q000494:pharmacology; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D000242:Cyclic AMP / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D005260:Female; D006062:Gonadotropins / Q000037:antagonists & inhibitors*; D015289:Leukotrienes / Q000494:pharmacology; D008054:Lipid Peroxides / Q000494:pharmacology*; D007986:Luteinizing Hormone / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D011374:Progesterone / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013256:Steroids / Q000037:antagonists & inhibitors*
D001555:Benzene Derivatives; D006062:Gonadotropins; D015289:Leukotrienes; D008054:Lipid Peroxides; D010618:Phenanthrolines; D013256:Steroids; C001635:ferroin; D011374:Progesterone; C025086:15-hydroperoxy-5,8,11,13-eicosatetraenoic acid; D007986:Luteinizing Hormone; D000242:Cyclic AMP; C007164:cumene hydroperoxide