Calcium pantothenate modulates gene expression in proliferating human dermal fibroblasts
Tonio Wiederholt
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Both authors contributed equally.
Search for more papers by this authorRuth Heise
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Both authors contributed equally.
Search for more papers by this authorClaudia Skazik
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorYvonne Marquardt
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorSylvia Joussen
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorKati Erdmann
College of Pharmacy, University of Minnesota, Minneapolis, MN, USA
Search for more papers by this authorHenning Schröder
College of Pharmacy, University of Minnesota, Minneapolis, MN, USA
Search for more papers by this authorHans F. Merk
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorJens Malte Baron
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorTonio Wiederholt
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Both authors contributed equally.
Search for more papers by this authorRuth Heise
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Both authors contributed equally.
Search for more papers by this authorClaudia Skazik
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorYvonne Marquardt
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorSylvia Joussen
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorKati Erdmann
College of Pharmacy, University of Minnesota, Minneapolis, MN, USA
Search for more papers by this authorHenning Schröder
College of Pharmacy, University of Minnesota, Minneapolis, MN, USA
Search for more papers by this authorHans F. Merk
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorJens Malte Baron
Department of Dermatology and Allergology, University Hospital of the RWTH, Aachen, Germany
Search for more papers by this authorAbstract
Abstract: Topical application of pantothenate is widely used in clinical practice for wound healing. Previous studies identified a positive effect of pantothenate on migration and proliferation of cultured fibroblasts. However, these studies were mainly descriptive with no molecular data supporting a possible model of its action. In this study, we first established conditions for an in vitro model of pantothenate wound healing and then analysed the molecular effects of pantothenate. To test the functional effect of pantothenate on dermal fibroblasts, cells were cultured and in vitro proliferation tests were performed using a standardized scratch test procedure. For all three donors analysed, a strong stimulatory effect of pantothenate at a concentration of 20 μg/ml on the proliferation of cultivated dermal fibroblasts was observed. To study the molecular mechanisms resulting in the proliferative effect of pantothenate, gene expression was analysed in dermal fibroblasts cultivated with 20 μg/ml of pantothenate compared with untreated cells using the GeneChip® Human Exon 1.0 ST Array. A number of significantly regulated genes were identified including genes coding for interleukin (IL)-6, IL-8, Id1, HMOX-1, HspB7, CYP1B1 and MARCH-II. Regulation of these genes was subsequently verified by quantitative real-time polymerase chain reaction analysis. Induction of HMOX-1 expression by pantothenol and pantothenic acid in dermal cells was confirmed on the protein level using immunoblots. Functional studies revealed the enhanced suppression of free radical formation in skin fibroblasts cultured with panthenol. In conclusion, these studies provided new insight in the molecular mechanisms linked to the stimulatory effect of pantothenate and panthenol on the proliferation of dermal fibroblasts.
References
- 1 Weimann B I, Hermann D. Studies on wound healing: effects of calcium D-pantothenate on the migration, proliferation and protein synthesis of human dermal fibroblasts in culture. Int J Vitam Nutr Res 1999: 69: 113–119.
- 2 Pugliese P T, Farina J C, Chautems Y. Effidacy of dexpanthenol in wound healing: double-blind assessment of excised wound tissue by ultrasound and histologic examination. Nouv Dermatol 1994: 14: 120–138.
- 3 Slyshenkov V S, Omelyanchik S N, Moiseenok A G, Petushok N E, Wojtczak L. Protection by pantothenol and beta-carotene against liver damage produced by low-dose gamma radiation. Acta Biochim Pol 1999: 46: 239–248.
- 4 Wojtczak L, Slyshenkov V S. Protection by pantothenic acid against apoptosis and cell damage by oxygen free radicals – the role of glutathione. Biofactors 2003: 17: 61–73.
- 5 Ebner F, Heller A, Rippke F, Tausch I. Topical use of dexpanthenol in skin disorders. Am J Clin Dermatol 2002: 3: 427–433.
- 6 Presto S, Wehmeyer A, Fibry A, Rippke F, Bielfeldt S. Stimulation of epidermal regeneration by 5% dexpanthenol – results of a placebo-controlled double-blind study. Z Hautkr 2001: 76: 86–90.
- 7 Watson A L, Fray T R, Bailey J, Baker C B, Beyer S A, Markwell P J. Dietary constituents are able to play a beneficial role in canine epidermal barrier function. Exp Dermatol 2006: 15: 74–81.
- 8 Biro K, Thaçi D, Ochsendorf F R, Kaufmann R, Boehncke W H. Efficacy of dexpanthenol in skin protection against irritation: a double-blind, placebo-controlled study. Contact Dermatitis 2003: 49: 80–84.
- 9 Proksch E, Nissen H F. Dexpanthenol enhances skin barrier repair and reduces inflammation after sodium lauryl sulphate-induced irritation. J Dermatol Treat 2002: 13: 173–178.
- 10 Lacroix B, Didier E, Grenier J F. Effects of pantothenic acid on fibroblastic cell cultures. Res Exp Med 1988: 188: 391–396.
- 11 Lacroix B, Didier E, Grenier J F. Role of pantothenic and ascorbic acid in wound healing processes: in vitro study on fibroblasts. Int J Vitam Nutr Res 1988: 58: 407–413.
- 12 Oztürk N, Korkmaz S, Oztürk Y. Wound-healing activity of St. John’s Wort (Hypericum perforatum L.) on chicken embryonic fibroblasts. J Ethnopharmacol 2007: 111: 33–39.
- 13 Klöcker N, Rudolph P, Verse T. Evaluation of protective and therapeutic effects of dexpanthenol on nasal decongestants and preservatives: results of cytotoxic studies in vitro. Am J Rhinol 2004: 18: 315–320.
- 14 Slyshenkov V S, Piwocka K, Sikora E, Wojtczak L. Pantothenic acid protects jurkat cells against ultraviolet light-induced apoptosis. Free Radic Biol Med 2001: 30: 1303–1310.
- 15 Vile G F, Basu-Modak S, Waltner C, Tyrrell R M. Heme oxygenase 1 mediates an adaptive response to oxidative stress in human skin fibroblasts. Proc Natl Acad Sci USA 1994: 29: 2607–2610.
- 16 Dewor M, Steffens G, Baron J M, Bernhagen J. Macrophage migration inhibitory factor (MIF) promotes fibroblast migration in wounded monolayers in vitro. FEBS Lett 2007: 581: 4734–4742.
- 17 Okoniewski M J, Hey Y, Pepper S D, Miller C J. High correspondence between Affymetrix exon and standard expression arrays. BioTechniques 2007: 42: 181–185.
- 18 Baron J M, Wiederholt T, Heise R, Merk H F, Bickers D R. Expression and function of Cytochrome p450-dependent enzymes in human skin cells. Curr Med Chem 2008: 15: 2258–2264.
- 19 Altschul S F, Gish W, Miller W, Myers E W, Lipman D J. Basic local alignment search tool. J Mol Biol 1990: 215: 403–410.
- 20 Skazik C, Heise R, Bostanci O et al. Differential expression of influx and efflux transport proteins in human antigen presenting cells. Exp Dermatol 2008: 17: 739–747.
- 21 Oberle S, Abate A, Grosser N et al. Heme oxygenase-1 induction may explain the antioxidant profile of pentaerithrityl trinitrate. Biochem Biophys Res Commun 2002: 290: 1539–1544.
- 22 Grosser N, Hemmerle A, Berndt G et al. The antioxidant defense protein heme oxygenase-1 is a novel target for statins in endothelial cells. Free Radic Biol Med 2004: 37: 2064–2071.
- 23 Erdmann K, Grosser N, Schipporeit K, Schröder H. The ACE inhibitory dipeptide Met-Tyr diminishes free radical formation in human endothelial cells via induction of heme oxygenase-1 and ferritin. J Nutr 2006: 136: 1–5.
- 24
Stütgen G,
Krause H.
Die percutane absorption von tritium–markiertem Panthenol bei Mensch und Tier.
Arch F Klin Exp Dermatol
1960: 209: 578–582.
10.1007/BF00477492 Google Scholar
- 25 Abiko Y, Tomikawa M, Shimizu M. Enzymatic conversion of pantothenylalcohol to pantothenic acid. J Vitaminol Kyoto 1969: 15: 59–69.
- 26 Lorenz K, Sicker M, Schmelzer E et al. Multilineage differentiation potential of human dermal skin-derived fibroblasts. Exp Dermatol 2008: 17: 925–932.
- 27 Burlet E. Die percutane resorption von panthenol (3-[2’, 4’-Dioxy-3’,3’dimethyl-butyryl]-aminopropanol). Z Vitaminforsch 1946: 16: 92–97.
- 28 Abe M, Yokoyama Y, Syuto T, Ishibuchi H, Ishikawa O. Interleukin-6 counteracts effects of cyclosporin A on extracellular matrix metabolism by human dermal fibroblasts. Cell Tissue Res 2008: 333: 281–288.
- 29 Luckett L R, Gallucci R M. Interleukin-6 (IL-6) modulates migration and matrix metalloproteinase function in dermal fibroblasts from IL-6KO mice. Br J Dermatol 2007: 156: 1163–1171.
- 30 Wang X P, Schunck M, Kallen K J et al. The interleukin-6 cytokine system regulates epidermal permeability barrier homeostasis. J Invest Dermatol 2004: 123: 124–131.
- 31 Gallucci R M, Sloan D K, Heck J M, Murray A R, O’Dell S J. Interleukin 6 indirectly induces keratinocyte migration. J Invest Dermatol 2004: 122: 764–772.
- 32 Takamiya M, Fujita S, Saigusa K, Aoki Y. Simultaneous detection of eight cytokines in human dermal wounds with a multiplex bead-based immunoassay for wound age estimation. Int J Legal Med 2008: 122: 143–148.
- 33 Kubo K, Kuroyanagi Y. A study of cytokines released from fibroblasts in cultured dermal substitute. Artif Organs 2005: 29: 845–849.
- 34 Dreuw A, Hermanns H M, Heise R et al. Interleukin-6-type cytokines upregulate expression of multidrug resistance-associated proteins in NHEK and dermal fibroblasts. J Invest Dermatol 2005: 124: 28–37.
- 35 Zheng W, Wang H, Xue L, Zhang Z, Tong T. Regulation of cellular senescence and p16(INK4a) expression by Id1 and E47 proteins in human diploid fibroblast. J Biol Chem 2004: 279: 31524–31532.
- 36 Clark J E, Green C J, Motterlini R. Involvement of the heme oxygenase-carbon monoxide pathway in keratinocyte proliferation. Biochem Biophys Res Commun 1997: 241: 215–220.
- 37 Hanselmann C, Mauch C, Werner S. Haem oxygenase-1: a novel player in cutaneous wound repair and psoriasis? Biochem J 2001: 353: 459–466.
- 38 Wagener F A, Van Beurden H E, Von Den Hoff J W, Adema G J, Figdor C G. The heme–heme oxygenase system: a molecular switch in wound healing. Blood 2003: 102: 521–528.
- 39 Becker J C, Grosser N, Schröder H, Domschke W, Pohle T. Gastroprotection by vitamin C – a heme oxygenase-dependent mechanism? Biochem Biophys Res Commun 2003: 312: 507–512.
- 40 Berndt G, Grosser N, Hoogstraate J, Schröder H. AZD3582 increases heme oxygenase-1 expression and antioxidant activity in vascular endothelial and gastric mucosal cells. Eur J Pharm Sci 2005: 25: 229–235.
- 41 Numata I, Shibahara S, Aiba S. The functional expression of heme oxygenase-1 in normal human epidermis. J Invest Dermatol 2008: 128: S101.
- 42 Maglara A, Vasilaki A, Jackson M J, McArdle A. Damage to developing mouse skeletal muscle myotubes in culture: protective effect of heat shock proteins. J Physiol 2003: 548: 837–846.
- 43 Reddi A S, Jyothirmayi G N, De Angelis B, Frank O, Baker H. Tissue concentration of water-soluble vitamins in normal and diabetic rats. Int J Vitr Nutr Res 1993: 63: 140–144.
- 44 Slyshenkov V S, Moiseenok A G, Wojtczak L. Noxious effects of oxygen reactive species on energy-coupling processes in Ehrlich ascites tumor mitochondria and the protection by pantothenic acid. Free Radic Biol Med 1996: 20: 793–800.
- 45 Moiseenok A G, Komar V I, Khormich T I, Kanunnikova N P, Slyshenkov V S. Pantothenic acid in maintaining thiol and immune homeostasis. BioFactors 2000: 11: 53–55.
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