bay-11-7082 and Vitiligo

Oxidative stress and autoimmune reaction are involved in the pathogenesis of vitiligo. Levels of hydrogen peroxide (H(2)O(2)) and interleukin-6 (IL-6), a proinflammation cytokine and a key factor in the pathogenesis of autoimmune diseases, have been reported to be elevated in vitiligo lesions. The objective of this study was to evaluate the effects of subtoxic levels of H(2)O(2) on the expression of IL-6 by cultured human epidermal melanocytes and to explore the relevant signal pathways. Cultured human melanocytes were stimulated with of H(2)O(2) at subtoxic levels. Levels of IL-6 protein in the medium and IL-6 mRNA in the cells were measured by IL-6 ELISA analysis and RT-PCR, respectively. NF-κB and phosphorylated p38 mitogen-activated protein kinase (MAPK), ERK and JNK in cells cultured with and without H(2)O(2) were measured by relevant ELISA kits. In cultured melanocytes, subtoxic levels of H(2)O(2) (30-300 μM) significantly increased the IL-6 mRNA and protein levels in a dose-dependent manner. NF-κB in nuclear extracts and phosphorylated p38 MAPK levels in cell lysates were significantly increased in H(2)O(2) treated cells. Pretreatment of cells with inhibitors of p38 MAPK (SB203580) and NF-κB (BAY11-7082), but not inhibitors of ERK (UO1026) and JNK (SP600125), abolished H(2)O(2)-induced expression of IL-6. H(2)O(2)-induced overexpression of IL-6 by melanocytes may be a molecular linkage for the oxidative stress and inflammatory/autoimmune reactions in vitiligo and may provide a novel target for the treatment of vitiligo.

Topics: Anthracenes; Cells, Cultured; Epidermis; Extracellular Signal-Regulated MAP Kinases; Humans; Hydrogen Peroxide; Imidazoles; Interleukin-6; Melanocytes; NF-kappa B; Nitriles; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Pyridines; Sulfones; Up-Regulation; Vitiligo

Intercellular adhesion molecule-1 (ICAM-1) expression has been detected in melanocytes around active vitiligo patches as well as in surgically transplanted melanocytes. However, it is unclear whether and how skin injury induces the inappropriate expression of ICAM-1 and other proinflammatory genes in melanocytes. We previously reported that human melanocytes expressed TLR3. We hypothesized that the TLR3 expressed in melanocytes may recognize skin injury by binding to the endogenous ligands secreted by the damaged keratinocytes. Here we showed that RNA released from necrotic keratinocytes induced the upregulation of ICAM-1 protein and mRNA, as shown by FACS and real-time RT-PCR. Use of NF-κB inhibitor prevents upregulation of ICAM-1 in melanocytes indicating a direct role of NF-κB in necrotic keratinocyte-mediated upregulation of ICAM-1. Using a shRNA-expressing lentivirus, we demonstrated that in human melanocytes, TLR3 seems to be necessary for the upregulation of ICAM-1. Using oligonucleotide microarray, we demonstrated a dramatic increase in proinflammatory cytokine and chemokine transcripts (CXCL10, CXCL11, TNFSF10, CCL5, CCL4, CCL2, IFNB1, CCL20, IL-8, and CCL11). These observations suggested that RNA released from necrotic keratinocytes might act as an endogenous TLR3 ligand for the stimulation of ICAM-1 and other proinflammatory gene expression in human melanocytes, which might be involved in the pathogenesis of vitiligo following skin physical trauma.

Topics: Cell Communication; Cytokines; Humans; Inflammation Mediators; Intercellular Adhesion Molecule-1; Keratinocytes; Melanocytes; Necrosis; NF-kappa B; Nitriles; Oligonucleotide Array Sequence Analysis; RNA; RNA, Small Interfering; Sulfones; Toll-Like Receptor 3; Ultraviolet Rays; Up-Regulation; Vitiligo