Prostaglandin E Receptor Subtypes in Cultured Rat Microglia and Their Role in Reducing Lipopolysaccharide‐Induced Interleukin‐1 β Production

Abstract

Abstract : Prostaglandins (PGs) are potent modulators of brain function under normal and pathological conditions. The diverse effects of PGs are due to the various actions of specific receptor subtypes for these prostanoids. Recent work has shown that PGE 2, while generally considered a proinflammatory molecule, reduces microglial activation and thus has an antiinflammatory effect on these cells. To gain further insight to the mechanisms by which PGE 2 influences the activation of microglia, we investigated PGE receptor subtype, i.e., EP1, EP2, EP3, and EP4, expression and function in cultured rat microglia. RT‐PCR showed the presence of the EP1 and EP2 but not EP3 and EP4 receptor subtypes. Sequencing confirmed their identity with previously published receptor subtypes. PGE 2 and the EP1 agonist 17‐phenyl trinor PGE 2 but not the EP3 agonist sulprostone elicited reversible intracellular [Ca 2+] increases in microglia as measured by fura‐2. PGE 2 and the EP2/EP4‐specific agonists 11‐deoxy‐PGE 1 and 19‐hydroxy‐PGE 2 but not the EP4‐selective agonist 1‐hydroxy‐PGE 1 induced dose‐dependent production of cyclic AMP (cAMP). Interleukin (IL)‐1β production, a marker of activated microglia, was also measured following lipopolysaccharide exposure in the presence or absence of the receptor subtype agonists. PGE 2 and the EP2 agonists reduced IL‐1β production. IL‐1β production was unchanged by EP1, EP3, and EP4 agonists. The adenylyl cyclase activator forskolin and the cAMP analogue dibutyryl cAMP also reduced IL‐1β production. Thus, the inhibitory effects of PGE 2 on microglia are mediated by the EP2 receptor subtype, and the signaling mechanism of this effect is likely via cAMP. These results show that the effects of PGE 2 on microglia are receptor subtype‐specific. Furthermore, they suggest that specific and selective manipulation of the effects of PGs on microglia and, as a result, brain function may be possible.