Role of ERK MAP kinases in responses of cultured human airway smooth muscle cells to IL-1 beta

We have previously reported that interleukin (IL)-1 beta causes beta-adrene rgic hyporesponsiveness in cultured human airway smooth muscle cells by inc reasing cyclooxygenase-2 (COX-2) expression and prostanoid formation. The p urpose of this study was to determine whether extracellular signal-regulate d kinases (ERKs) are involved in these events. Levels of phosphorylated ERK (p42 and p44) increased 8.3- and 18-fold, respectively, 15 min after treat ment with IL-1 beta (20 mg/ml) alone. Pretreating cells with the mitogen-ac tivated protein kinase kinase inhibitor PD-98059 or U-126 (2 h before IL-1 beta treatment) decreased ERK phosphorylation. IL-1 beta (20 ng/ml for 22 h ) alone caused a marked induction of COX-2 and increased basal PGE(2) relea se 28-fold (P < 0.001). PD-98059 (100 mu M) and U-126 (10 mu M) each decrea sed COX-2 expression when administered before IL-1 beta treatment. In contr ol cells, PD-98059 and U-126 had no effect on basal or arachidonic acid (AA ; 10 mu M)-stimulated PGE(2) release, but both inhibitors caused a signific ant decrease in bradykinin (BK; 1 mu M)-stimulated PGE(2) release, consiste nt with a role for ERK in the activation of phospholipase A(2) by BK. In IL -lp-treated cells, prior administration of PD-98059 caused 81, 92 and 40% d ecreases in basal and BK-and AA-stimulated PGE(2) release, respectively (P < 0.01), whereas administration of PD-98059 20 h after IL-1 beta resulted i n only 38 and 43% decreases in basal and BK-stimulated PGE(2) release, resp ectively (P < 0.02) and had no effect on AA-stimulated PGE(2) release. IL-1 beta attenuated isoproterenol-induced decreases in human airway smooth mus cle stiffness as measured by magnetic twisting cytometry, and PD-98059 or U -126 abolished this effect in a concentrationdependent manner. These result s are consistent with the hypothesis that ERKs are involved early in the si gnal transduction pathway through which IL-1 beta induces PGE(2) synthesis and beta-adrenergic hyporesponsiveness and that ERKs act by inducing COX-2 and activating phospholipase A(2).