Objective: To study the effect of nitric oxide (NO) on atrial natriuretic peptide (ANP) and progesterone (P) production by human granulose luteinized cells (GLC) in vitro and to elucidate their role on the survival of cultured cells.
Methods: Human GLCs were cultured in HAM's F10/10% FCS as monolayers for 24 h. Subsequently GLCs were treated for 24 h with 0.5 mM Sodium Nitroprusside (SNP, NO donor) and 0.5 mM Aminoglutethimide (AG , P450scc inhibitor). The levels of ANP and P were measured in supernatants of cultured cells by proANP(1-98) kit and RIA, respectively. Caspase-3 activity was determined by Ac-DEVD-pNA as substrate.
Results: The production of ANP and P was increased by NO as compared to control cells (p<0.05). AG diminished the production of P compared to SNP (p<0.05). The caspase-3 activity was significantly lower in SNP treated cells (p<0.05) and increased significantly after AG treatment compared to control cells (p<0.05).
Conclusion: NO generated by SNP in human GLCs culture stimulated the production of ANP and P. The higher levels of ANP and P were closely related to significantly lower caspase-3 activity thus showing the role of ANP, P and NO on the survival of preovulatory human follicle.