α-Melanocyte-Stimulating Hormone Inhibits the Nuclear Transcription Factor NF-κB Activation Induced by Various Inflammatory Agents

Abstract α-Melanocyte-stimulating hormone (α-MSH) is a tridecapeptide found mainly in the brain, pituitary, and circulation. It inhibits most forms of inflammation by a mechanism that is not known. As most types of inflammation require activation of NF-κB, we investigated the effect of α-MSH on the activation of this transcription factor by a wide variety of inflammatory stimuli. Electrophoretic mobility shift assay showed that α-MSH completely abolished TNF-mediated NF-κB activation in a dose- and time-dependent manner. It also suppressed NF-κB activation induced by LPS, okadaic acid, and ceramide. The effect was specific, as the activation of the transcription factor activating protein-1 by TNF was unaffected. Western blot analysis revealed that TNF-dependent degradation of the inhibitory subunit of NF-κB, IκBα, and nuclear translocation of the p65 subunit of NF-κB were also inhibited. This correlated with suppression of NF-κB-dependent reporter gene expression induced by TNF. The inhibitory effect of α-MSH appeared to be mediated through generation of cAMP, as inhibitors of adenylate cyclase and of protein kinase A reversed its inhibitory effect. Similarly, addition of membrane-permeable dibutyryl cAMP, like α-MSH, suppressed TNF-induced NF-κB activation. Overall, our results suggest that α-MSH suppresses NF-κB activated by various inflammatory agents and that this mechanism probably contributes to its anti-inflammatory effects.