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Antisteroidogenic action of nitric oxide on human corpus luteum in vitro

Mechanism of action

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Abstract

To analyze the mechanism by which nitric oxide (NO) exerts its antisteroidogenic action, human luteal cells were cultured during 24 and 48 h with l-arginine (l-Arg, 1 mmol/L); 1,2(2-trifluoromethylphenyl)imidazole (TRIM) (50 μmol/L and 1 mmol/L) and cyclic guanosine monophosphate (cGMP) analog (8-Br-cGMP, 1 mmol/L). Estradiol, nitrite, and P450 AROM activity were determined in culture media. Total cGMP concentration was evaluated in the cells and culture media by radioimmunoassay, and NADPH diaphorase was used as a histochemical marker for NO synthase (NOS) activity. During the corpus luteum (CL) life-span, NO affected estradiol secretion in an age-dependent manner, with an inhibition in mid-CL (37%; p<0.05) in agreement with our previous results, and no significant modification in early and late CL. Basal nitrite concentration in 24 and 48 h of midluteal cell cultures (42 and 93 pmol/106 cells, respectively) was increased by l-Arg (53% and 88%) and inhibited by the two TRIM concentrations; also, an intense diaphorase reactivity was observed in endothelial cells and luteal parenchyma. Total cGMP was not detected in cell cultures and 8-Br-cGMP did not modify estradiol secretion, whereas aromatase activity was strongly inhibited by l-Arg (70%, p<.05). These results suggest that both NOS isoforms are active in midluteal cells, and the mechanism of action for NO on in vitro estradiol secretion may be an inhibition of P450 AROM activity.

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Authors and Affiliations

  1. Department of Biology, School of Science, University of Valparaiso, Valparaiso, Chile

    Hugo A. Diaz

  2. Postdoctoral Fellow (PLACIRH), School of Medicine, University of Chile, Chile

    Carlos Stocco

  3. Reproductive Medicine and Infertility, School of Medicine, University of Chile, Chile

    Alberto Palomino

  4. Institute of Maternal and Child Research, University of Medicine, School of Medicine, P.O. Box 226-3, Santiago, Chile

    M. Cecilia Johnson, Hugo A. Diaz, Carlos Stocco, Alberto Palomino, Luigi Devoto &  Margarita Vega

Authors
  1. M. Cecilia Johnson
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  2. Hugo A. Diaz
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  3. Carlos Stocco
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  4. Alberto Palomino
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  5. Luigi Devoto
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  6. Margarita Vega
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Correspondence to Margarita Vega.

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Johnson, M.C., Diaz, H.A., Stocco, C. et al. Antisteroidogenic action of nitric oxide on human corpus luteum in vitro. Endocr 11, 31–36 (1999). https://doi.org/10.1385/ENDO:11:1:31

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  • DOI: https://doi.org/10.1385/ENDO:11:1:31

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