ALPHA-MELANOCYTE-STIMULATING HORMONE INDUCES COLLAGENASE MATRIX METALLOPROTEINASE-1 IN HUMAN DERMAL FIBROBLASTS

Recent reports have suggested that alpha-melanocyte stimulating hormone (alpha-MSH) plays an important role in ultraviolet (UV) irradiation mediated skin changes including pigmentation, inflammation and connective tissue damage, alpha-MSH synthesis has been found to be induced in human keratinocytes following UV irradiation. In order to test the hypothesis whether UV induced alpha-MSH - via a paracrine loop - regulates the synthesis and the activity of collagenase/MMP-1, we studied the effects of alpha-MSH on the expression of MMP-1 and its tissue inhibitor of matrix metalloproteinases (TIMP-1). Confluent human dermal fibroblast cultures from foreskin biopsies of healthy human donors were treated with 10(-5) M, 10(-8) M and 10(-1) M alpha-MSH for 30 min, As determined by Northern blot analysis 10(-5) M and 10(-8) M alpha-MSH dose- and time-dependently induced steady state levels of MMP-1 mRNA up to 9-fold compared to untreated controls, TIMP-1 mRNA steady state levels were also slightly induced, however, the increased MMP-1/TIMP-1 ratio when normalized to beta-actin reflected an unbalanced synthesis. MMP-1 protein expression was studied with an immunofluorescence technique using a monoclonal antibody against MMP-1. After alpha-MSH treatment an increased number of fibroblasts revealed an intense perinuclear staining pattern compared to the less intense staining of control fibroblasts. The overall collagenolytic activity of supernatants from alpha-MSH treated fibroblasts was increased by 35%. Our data support the view that UV induced alpha-MSH - by the stimulation of collagenase/MMP-1 may contribute to the loss of interstitial collagen related to cutaneous photoaging.