Activation of the p38 and p42/p44 mitogen-activated protein kinase families by the histamine H1 receptor in DDT1MF-2 cells

摘要

The mitogen-activated protein kinases (MAPKs) consist of the p42/p44 MAPKs and the stress-activated protein kinases, c-Jun N-terminal kinase (JNK) and p38 MAPK. In this study we have examined the effect of histamine H1 receptor activation on MAPK pathway activation in the smooth muscle cell line DDT1MF-2.Histamine stimulated time and concentration-dependent increases in p42/p44 MAPK activation in DDT1MF-2 cells. Responses to histamine were inhibited by the histamine H1 receptor antagonist mepyramine (KD 3.5 nM) and following pre-treatment with pertussis toxin (PTX; 57% inhibition).Histamine-induced increases in p42/p44 MAPK activation were blocked by inhibitors of MAPK kinase 1 (PD 98059), tyrosine kinase (genistein and tyrphostin A47), phosphatidylinositol 3-kinase (wortmannin and LY 294002) and protein kinase C (Ro 31-8220; 10 μM; 41% inhibition). Inhibitors of Src tyrosine kinase (PP2) and the epidermal growth factor tyrosine kinase (AG1478) were without effect. Removal of extracellular Ca2+, chelation of intracellular Ca2+ with BAPTA and inhibition of focal adhesion assembly (cytochalasin D) had no significant effect on histamine-induced p42/p44 MAPK activation.Histamine stimulated time and concentration-dependent increases in p38 MAPK activation in DDT1MF-2 cells but had no effect on JNK activation. Histamine-induced p38 MAPK activation was inhibited by pertussis toxin (74% inhibition) and the p38 MAPK inhibitor SB 203580 (95% inhibition).In summary, we have shown the histamine H1 receptor activates p42/p44 MAPK and p38 MAPK signalling pathways in DDT1MF-2 smooth muscle cells. Interestingly, signalling to both pathways appears to involve histamine H1 receptor coupling to Gi/Go-proteins.