Lipopolysaccharide induces the expression of prostaglandin H synthase-2 with subsequent production of PGE2 in human microvascular endothelial cells. In order to investigate the signaling pathways that are involved in the LPS-induced production of PGE2, the roles of several second messenger systems in this process were evaluated. Treatment of human microvascular endothelial cells with the tyrosine kinase inhibitors herbimycin A and genistein significantly inhibited LPS-mediated PGE2 production. Western blotting demonstrated that LPS induced the phosphorylation of p42, p44 and p38 MAP kinases. Both PD98059, an inhibitor of p42 and p44 MAP kinase activation, and SB203580, an inhibitor of p38 MAP kinase, blocked LPS-induced PGE2 production. Inhibition of PKC activity by staurosporine and GF109203X resulted in a potentiation of the LPS response. Treatment of the cells with SN50, an NF-κB inhibitor, resulted in inhibition of the LPS-mediated PGE2 production. These results suggest that LPS induces PGE2 production in human microvascular endothelial cells through sequential activation of tyrosine kinases, ERK and p38 MAP kinases and NF-κB. In addition, PKC activity exerts a feedback control of PGE2 production in LPS-stimulated microvessel-derived endothelial cells.
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