Atrial natriuretic peptide (ANP) contributes to the inhibition of such causes of inflammation as the lipopolysaccharide (LPS)-induced productions of nitric oxide (NO) and proinflammatory cytokines [including interleukin-1 (IL-1)] in macrophages. In the present study we used primary cultures of rat brain macrophage-like cells (i.e., microglial cells) to investigate whether ANP binding to its receptors inhibits LPS-induced microglial activation via effects on the activation of the proinflammatory transcription factors NF-kappaB and AP-1. The productions of NO and IL-1, as well as morphological changes, were examined to assess LPS-induced activation of microglial cells. Our RT-PCR study revealed that rat microglial cells express the mRNAs for ANP receptors (types A, B, and C) and that for the ANP molecule. LPS (100 ng/ml)-stimulated microglial cells showed increases in nitrite (a relatively stable metabolite of NO) and IL-1 concentrations, and in the expression of IL-1 mRNA, as well as a morphological change from an amoeboid shape to a multipolar (mostly bipolar, but sometimes tripolar) rod shape. These effects were all significantly inhibited by treatment with ANP (at 10(-6)M or less). The inhibition by ANP of the LPS-induced nitrite response was abrogated by a NP-receptor antagonist, HS-142-1 (100 ng/ml). NF-kappaB and AP-1 activities were enhanced in LPS-stimulated microglial cells, and these enhancements were significantly suppressed by ANP (10(-6)M). These results suggest that ANP inhibits LPS-stimulated activities in microglial cells through activation of microglial ANP receptors, leading to inhibitions of NF-kappaB and AP-1.