The Effect of Nitric Oxide on Apoptosis in Human Luteinized Granulosa Cells / 대한산부인과학회잡지

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.