title: Hyaluronic acid abrogates ethanol-dependent inhibition of collagen biosynthesis in cultured human fibroblasts.
abstract: BACKGROUND
  The aim of the study was to evaluate the effect of ethanol on collagen biosynthesis in cultured human skin fibroblasts, and the role of hyaluronic acid (HA) in this process. Regarding the mechanism of ethanol action on human skin fibroblasts we investigated: expression of β1 integrin and insulin-like growth factor 1 receptor (IGF-IR), signaling pathway protein expression: mitogen-activated protein kinases (MAPKs), protein kinase B (Akt), nuclear factor kappa B (NF-κB) transcription factor, cytotoxicity assay and apoptosis, metalloproteinase activity, as well as the influence of HA on these processes.
  
  
  METHODS
  Collagen biosynthesis, activity of prolidase, DNA biosynthesis, and cytotoxicity were measured in confluent human skin fibroblast cultures that have been treated with 25, 50, and 100 mM ethanol and with ethanol and 500 µg/mL HA. Western blot analysis and zymography were performed to evaluate expression of collagen type I, β1 integrin receptor, IGF-IR, NF-κB protein, phospho-Akt protein, kinase MAPK, caspase 9 activity, and matrix metalloproteinases (MMP-9 and MMP-2).
  
  
  RESULTS
  Ethanol in a dose-dependent manner lead to the impairment of collagen biosynthesis in fibroblast cultures through decreasing prolidase activity and expression of β1 integrin and IGF-IR. This was accompanied by an increased cytotoxicity, apoptosis and lowered expression of the signaling pathway proteins induced by β1 integrin and IGF-IR, that is, MAPK (ERK1/2) kinases. The lowered amount of synthesized collagen and prolidase activity disturbance may also be due to the activation of NF-κB transcription factor, which inhibits collagen gene expression. It suggests that the decrease in fibroblast collagen production may be caused by the disturbance in its biosynthesis but not degradation. The application of HA has a protective effect on disturbances caused by the examined substances. It seems that regulatory mechanism of ethanol-induced collagen aberration take place at the level of collagen biosynthesis, since no effect of ethanol and HA was found on process of collagen degradation by MMP-2 and MMP-9.
  
  
  CONCLUSIONS
  This study provides evidence that ethanol impairs collagen metabolism in human skin fibroblasts, leading to a significant decrease in the amount of produced protein. This mechanism probably is due to downregulation of prolidase activity, expression of β1 integrin and IGF-IR receptors, and the signaling pathway proteins induced by these receptors.
authors.affiliation: Department of Esthetic Medicine, Medical University of Białystok, Białystok, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Donejko
authors.affiliation: Department of Esthetic Medicine, Medical University of Białystok, Białystok, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Przylipiak
authors.affiliation: Department of Medicinal Chemistry, Medical University of Białystok, Białystok, Poland.
authors.forename: Edyta
authors.initials: E
authors.lastname: Rysiak
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Medical University of Białystok, Białystok, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Miltyk
authors.affiliation: Department of Esthetic Medicine, Medical University of Białystok, Białystok, Poland.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Galicka
authors.affiliation: Medical Office, Białystok, Poland.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Przylipiak
authors.affiliation: Department of Medicinal Chemistry, Medical University of Białystok, Białystok, Poland.
authors.forename: Ilona
authors.initials: I
authors.lastname: Zaręba
authors.affiliation: Department of Medicinal Chemistry, Medical University of Białystok, Białystok, Poland.
authors.forename: Arkadiusz
authors.initials: A
authors.lastname: Surazynski
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis* / Q000378:metabolism; D004150:Dipeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D019012:Integrin beta1 / Q000378:metabolism; D017526:Receptor, IGF Type 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship
chemical_list: D019012:Integrin beta1; D000431:Ethanol; D006820:Hyaluronic Acid; D003094:Collagen; D017526:Receptor, IGF Type 1; D004150:Dipeptidases; C022449:proline dipeptidase
keywords: collagen; ethanol; fibroblast; hyaluronic acid