Modulatory effects of progesterone on inducible nitric oxide synthase expression in vivo and in vitro

Nitric oxide (NO) is produced in the CNS following injury-induced expression of inducible nitric oxide synthase (iNOS), yet its role as protective or damaging is unclear. Previous studies investigating the therapeutic potential of female sex steroids in stroke and trauma suggest that NO from this source is harmful, since oestradiol and progesterone decreased the level of iNOS expression in vitro and improved neurological outcome. We investigated the effects of progesterone on stroke-induced expression of iNOS in mice, as well as cytokine-induced expression of iNOS and its transcriptional activators in cells relevant to injury. We observed a significant reduction in stroke-induced iNOS transcript in progesterone-treated mice and in cultured macrophages. In contrast, progesterone significantly amplifed cytokine-induced iNOS mRNA in cultured primary astrocytes, although the expression of protein was decreased. We sequenced upstream of the 1.5 kb reported iNOS promoter region and identified a potential progesterone response element (PRE). Astrocytes transiently transfected with iNOS promoter/CAT reporter gene constructs containing the PRE displayed a significant increase in induction of CAT expression after progesterone treatment, and this was diminished in cells transfected with a construct containing a disrupted PRE. These observations suggest the involvement of iNOS in the neuroprotective effects of progesterone.