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Korean Journal of Obstetrics & Gynecology 2002;45(5):766-773.
Published online May 1, 2002.
The Effect of Nitric Oxide on Apoptosis in Human Luteinized Granulosa Cells.
Byung Chul Jee, Shin Yong Moon
1Department of Obstetrics and Gynecology, College of Medicine, Cheju National University, Cheju, Korea.
2Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Seoul National University, Seoul, Korea.
3Department of Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea.
Abstract
OBJECTIVE
To investigate the effect of nitric oxide on the apoptosis of human luteinized granulosa cells. METHODS : Granulosa cell suspensions were incubated for 48 hours after adding nitric oxide donor (S-nitroso-N-acetyl-penicillamine, SNAP) and nitric oxide synthase inhibitor (nitro-L-arginine methyl ester, L-NAME) at different concentrations. Apoptosis was examined using a terminal deoxynucleotide transferase- mediated dUTP-biotin nick end labeling method, and immunocytochemical staining was performed for six apoptosis-related proteins. RESULTS: Apoptotic rates were significantly lower in cells incubated in SNAP 0.5 mM, but higher in L-NAME 0.5, 1.0, and 5.0 mM. SNAP 0.5 mM lowered the expressions of Fas and p53 in granulosa cells, but Bcl-2 expression was increased, and Fas ligand or Bax remained unchanged. In L-NAME 0.5 and 5.0 mM, the expressions of p53 and Bax were increased, and Bcl-2 was unchanged. Fas/Fas ligands were also activated especially in L-NAME 5.0 mM. CONCLUSION: Nitric oxide may inhibit apoptosis via decreased Fas and p53, and increased Bcl-2 expression in human luteinized granulosa cells.
Key Words: Nitric oxide, Apoptosis, Human luteinized granulosa cell


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