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J. Biol. Chem., Vol. 275, Issue 21, 15629-15636, May 26, 2000
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alpha -Melanocyte-stimulating Hormone Reduces Impact of Proinflammatory Cytokine and Peroxide-generated Oxidative Stress on Keratinocyte and Melanoma Cell Lines*

John W. HaycockDagger §, Sarah J. RoweDagger , Susan Cartledge, Alice WyattDagger , Ghanem Ghanem||, Ronalto Morandini||, Ian G. Rennie**, and Sheila MacNeilDagger

From the Dagger  University Section of Medicine, Division of Clinical Sciences, Northern General Hospital, Sheffield S5 7AU, United Kingdom,  Department of Clinical Chemistry, Northern General Hospital, Sheffield S5 7AU, United Kingdom, || Laboratory of Oncology and Experimental Surgery, Institut Bordet, Université Libre de Bruxelles, B-1000 Brussels, Belgium, and the ** University Department of Opthalmology and Orthoptics, Royal Hallamshire Hospital, Sheffield, S10 2JF, United Kingdom

We have previously shown that alpha -melanocyte-stimulating hormone (alpha -MSH) can oppose tumor necrosis factor alpha  activation of NF-kappa B (1-2 h) and intercellular adhesion molecule 1 up-regulation (mRNA by 3 h and protein by 24 h) in melanocytes and melanoma cells. The present study reports on the ability of four MSH peptides to control intracellular peroxide levels and glutathione peroxidase (GPx) activity in pigmentary and nonpigmentary cells. In human HBL melanoma and HaCaT keratinocytes tumor necrosis factor alpha  and H2O2 both activated GPx in a time- and concentration-dependent manner (by 30-45 min). alpha -MSH peptides were found to inhibit the stimulated GPx activity and had biphasic dose-response curves. MSH 1-13 and MSH [Nle4-D-Phe7] achieved maximum inhibition at 10-10 and 10-12 M, respectively. Higher concentrations (10-100 fold) of MSH 4-10 and MSH 11-13 were required to produce equivalent levels of inhibition. alpha -MSH was also capable of reducing peroxide accumulation within 15 min, and again this inhibition was biphasic. The data support a role of alpha -MSH in acute protection of cells to oxidative/cytokine action that precedes NF-kappa B and GPx activation. The rapidity and potency of the response to alpha -MSH in pigmentary and nonpigmentary cells suggest this to be a central role of this peptide in cutaneous cells.

* This work was supported by registered Charity No. 226678, Grant 87762 from the Special Trustees for the Former United Sheffield Hospitals, Sheffield, UK.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed. Tel.: 44-114-2714007; Fax: 44-114-2560458; E-mail: j.w.haycock@shef.ac.uk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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