JNK activation is essential for activation of MEK/ERK signaling in IL-1β-induced COX-2 expression in synovial fibroblasts

The proinflammatory cytokine interleukin 1 beta(IL-1 beta) induces prostaglandin E-2 (PGE(2)) production via upregulation of cyclooxygenase-2 (COX-2) expression in synovial fibroblasts. This effect of IL-1 beta is involved in osteoarthritis. We investigated MAPK signaling pathways in IL-1 beta-induced COX-2 expression in feline synovial fibroblasts. In the presence of MAPK inhibitors, IL-1 beta-induced COX-2 expression and PGE(2) release were both attenuated. IL-1 beta induced the phosphorylation of p38, JNK, MEK, and ERK1/2. A JNK inhibitor prevented not only JNK phosphorylation but also MEK and ERK1/2 phosphorylation in IL-1 beta-stimulated cells, but MEK and ERK1/2 inhibitors had no effect on JNK phosphorylation. A p38 inhibitor prevented p38 phosphorylation, but had no effect on MEK, ERK1/2, and JNK phosphorylation. MEK, ERK1/2, and JNK inhibitors had no effect on p38 phosphorylation. We also observed that in IL-1 beta-treated cells, phosphorylated MEK, ERK1/2, and JNK were co-precipitated with anti-phosphoMEK, ERK1/2, and JNK antibodies. The silencing of JNK1 in siRNA-transfected fibroblasts prevented IL-1 beta to induce phosphorylation of MEK and ERK1/2 and COX-2 mRNA expression. These observations suggest that JNK1 phosphorylation is necessary for the activation of the MEK/ERK1/2 pathway and the subsequent COX-2 expression for PGE(2) release, and p38 independently contributes to the IL-1 beta effect in synovial fibroblasts.