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The present study was undertaken to determine whether NO involved in the regulation of ovarian steroidogenesis in female rats. The purpose of the present study was to investigate whether NO might inhibits ovarian steroidogenesis. Female rats were divided of five rats including, receiving, N-omega-nitro-L-arginine methyle ester (L-NAME), an inhibitor of NO synthesis, Trinitroglycerin (TNG), an NO donor, L-arginine, normal saline. After 21 days, concenteration of progesterone and oesteradiol were measured by Electerochemiluminescence (ECL). Result showed that: TNG significantly decreased concentration of progesterone (50%) and L-NAME partially increased ovarian oesteradiol, but not differ effect from that rats treated with trinitroglycerin.