Novel sphingolipid derivatives promote keratinocyte differentiation
György Paragh
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Department of Dermatology, Semmelweis University, Budapest, Hungary
Department of Medical Chemistry, Semmelweis University, Budapest, Hungary
Search for more papers by this authorPetra Schling
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorPeter Ugocsai
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorGerhard Liebisch
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorSusanne Heimerl
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorChristoph Moehle
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorNorbert M. Wikonkál
Department of Dermatology, Semmelweis University, Budapest, Hungary
Search for more papers by this authorJózsef Mandl
Department of Medical Chemistry, Semmelweis University, Budapest, Hungary
Search for more papers by this authorThomas Langmann
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Institute of Human Genetics, University of Regensburg, Germany
Search for more papers by this authorGerd Schmitz
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorGyörgy Paragh
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Department of Dermatology, Semmelweis University, Budapest, Hungary
Department of Medical Chemistry, Semmelweis University, Budapest, Hungary
Search for more papers by this authorPetra Schling
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorPeter Ugocsai
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorGerhard Liebisch
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorSusanne Heimerl
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorChristoph Moehle
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorNorbert M. Wikonkál
Department of Dermatology, Semmelweis University, Budapest, Hungary
Search for more papers by this authorJózsef Mandl
Department of Medical Chemistry, Semmelweis University, Budapest, Hungary
Search for more papers by this authorThomas Langmann
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Institute of Human Genetics, University of Regensburg, Germany
Search for more papers by this authorGerd Schmitz
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany
Search for more papers by this authorAbstract
Abstract: Sphingolipids are important components of the water permeability barrier of the skin. Moreover, ceramides were also shown to influence keratinocyte differentiation and regulate cellular signalling. A confluence-induced differentiation model of normal human keratinocytes was established to allow evaluation of pro- and anti-differentiation effects of exogenous compounds. The effects of phytosphingosine (PS), sphingosine (SO), sphinganine (SA) and their hexanoyl (–C6), stearoyl (–C18) and salicyl (–SLC) derivatives, C12-alkylamine-salicylate (C12-SLC), salicylate (SLC) along with vitamin D3 (VD3) and retinol as control substances were tested in this system. Cytotoxicity assays were carried out to optimize the incubation conditions of compounds and whole genome expression changes were monitored by DNA-microarray on days 0, 1 and 4. Geometric means of gene expression levels of a subset of known keratinocyte differentiation-related genes were calculated from the microarray data to compare effects of the sphingolipid derivatives. Compound treatment-induced transcriptional changes were analysed by the ExPlain™ software (BIOBASE GmbH). Five of the assayed substances (SA, SO-C6, PS-C6, SO-SLC, PS-SLC) were found to be potent promoters of keratinocyte differentiation compared with VD3, and C12-SLC revealed potential anti-differentiation properties. ExPlain™ analysis found a different regulatory profile in the computed transcriptional networks of the sphingoid bases versus their –C6 and especially –SLC derivatives suggesting that the change in their keratinocyte differentiation modifying potential is due to a unique effect of the covalent attachment of the salicylic acid. Taken together, these results demonstrate the gene regulatory potential of sphingolipid species that could be valuable for dermatological or cosmetic applications.
Supporting Information
Figure S1. Sphingolipid compound treatment induces similar changes in gene expression measured by both microarray and TaqMan® quantitative PCR. Data represent gene expression values (mean +SD) of glutaredoxin, kallikrein 5, kallikrein 6, kallikrein 7 and UDP-glucose ceramide glucosyltransferase 1 day after compound treatment. Quantitative PCR results were normalized to 18S RNA expression values, and all expression changes are presented relative to day 1 controls.
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