Expression of inducible nitric oxide synthase during rat brain inflammation: regulation by 1,25-dihydroxyvitamin D3

This study, based on in situ hybridization and immunolabeling experiments, presents the time-course and cellular distribution of inducible NO synthase (iNOS) expression in a rat model of brain inflammation. Both intrahippocampal injection of lipopolysaccharide (LPS) or of buffer (stab lesion) induce an early, transient, and restricted expression of iNOS mRNA and immunoreactivity in the rat CNS. The topographic and phenotypic characteristics of iNOS-producing cells are distinct. After stab lesion, iNOS mRNAs, expressed at 5 h mainly in cells in the interventricular junction and in a few cells in brain parenchyma, were no more detectable from 15 h onwards, whereas the protein was faintly expressed in parenchymal cells at 15 h and 24 h. In contrast, after LPS injection, iNOS-mRNAs were detected from 5 to 24 h. iNOS-immunoreactivity was highly induced and sequentially observed first in choroid plexus and ependymal cells at 5 h, in monocytes and activated/reactive microglia at 15 h and 24 h, and finally in astrocytes at 72 h. In order to investigate potential regulatory effects of 1,25-dihydroxyvitamin D3 (1,25-D3) on iNOS expression, we have delivered this hormone with LPS or buffer into the rat hippocampus. 1,25-D3 significantly inhibits iNOS expression, at both the mRNA and immunoreactive protein levels, 15 h and 24 h after LPS injection, in the cells of the monocyte lineage. Moreover, 72 h after LPS injection, the addition of 1,25-D3 leads to a 6-fold increase in the number of macrophages around the lesion site, that correlates with a decrease in the proportion of apoptotic cells. Since 1,25-D3 can be produced by activated macrophages/microglia and since NO stimulates 1,25-D3 synthesis by macrophages, our results support the hypothesis that this hormone might be synthesized endogenously during CNS inflammatory reactions, thus explaining the transient and restricted iNOS expression observed after LPS intracerebral injection.