Direct Effects of Cutaneous Neuropeptides on Adenylyl Cyclase Activity and Proliferation in a Keratinocyte Cell Line: Stimulation of Cyclic AMP Formation by CGRP and VIP/PHM, and Inhibition by NPY Through G Protein-Coupled Receptors

Abstract

Many neuropeptides are present in the peripheral nerves of human skin and are distributed from the intraepidermis to subcutaneous appendages, and those peptides are considered to be involved in the pathogenesis of various inflammatory dermatoses. In this investigation, we determined the effects of various neuropeptides on intracellular cyclic adenosine-5'-monophosphate (AMP) formation in cultured human keratinocytes. Among the many peptides tested, calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), peptide histidine-methionine (PHM), and growth hormone releasing factor (GRF) stimulated a rapid and marked formation of intracellular cyclic AMP in keratinocytes in a dose-dependent manner. The direct association of the receptors for CGRP and VIP with adenylyl cyclase in keratinocytes was confirmed by the findings that CGRP and VIP stimulated the enzyme activity in membrane preparations derived from cultured keratinocytes in the presence of guanosine triphosphate (GTP). On the other hand, neuropeptide Y (NPY) showed an inhibitory effect on forskolin-induced cyclic AMP accumulation in keratinocytes. This inhibitory effect of NPY was completely eliminated by glucocorticoid pretreatment of cultured keratinocytes. Furthermore, the presence of peptides that substantially increase intracellular cyclic AMP accumulation also stimulated DNA synthesis and proliferation in a human keratinocyte cell line in a dose-dependent manner. These results suggest that neuropeptides work directly as biologic modulators of keratinocytes through the cyclic AMP cascade.