Progesterone inhibits superoxide radical production by mononuclear phagocytes in pseudopregnant rats

This study investigated the changes in superoxide radical production by mononuclear phagocytes in the corpus luteum (CL) during pseudopregnancy in rats. Activity of superoxide radical production was determined by the conversion of nitro blue tetrazolium (NBT) to blue formazan deposit. Rats received 10 mg NBT via the abdominal aorta on day 3, 7, or 13 of pseudopregnancy and were autopsied 1 min later to prepare the histological sections. The cells with blue formazan deposits (NBT-positive cells) in the CL were scarce on days 3 and 7 of pseudopregnancy and significantly increased on day 13 of pseudo-pregnancy. On the other hand, simultaneous administration of 100 micrograms phorbol 12-myristate 13-acetate, which activates mononuclear phagocytes to produce superoxide radical, significantly increased the numbers of NBT-positive cells in the CL on day 7 of pseudopregnancy, but not in the CL on day 3 or 13 of pseudopregnancy. To study the possibility that superoxide radical production by mononuclear phagocytes is inhibited by progesterone on day 7 of pseudopregnancy, peritoneal mononuclear phagocytes prepared on day 7 or 13 of pseudopregnancy were preincubated with 10, 50, or 100 ng/ml progesterone for 6 h and then stimulated with phorbol 12-myristate 13-acetate. Superoxide radical production was measured by the cytochrome c reduction method. One hundred nanograms per ml progesterone significantly inhibited superoxide radical production by mononuclear phagocytes, and this inhibitory effect of progesterone was significantly blocked by the simultaneous addition of RU486 (10(-7) M). These results suggested that progesterone inhibited superoxide radical production by the mononuclear phagocytes in the CL during midpseudopregnancy in rats.