title: The mechanism of oxythiamine-induced collagen biosynthesis in cultured fibroblasts.
abstract: The oxythiamine (OXY) is antivitamin of thiamine. The finding that OXY increases the cytoplasmic concentration of pyruvate, known to enhance collagen biosynthesis, led us to investigate the mechanism of this antivitamin action on collagen biosynthesis in cultured human skin fibroblasts. Confluent fibroblasts were treated with micromolar concentrations (30-1,000 µM) of OXY for 24 and 48 h. It was found that OXY-dependent increase in collagen biosynthesis was accompanied by parallel increase in prolidase activity and level, compared to untreated cells. Since phosphoenolpyruvate (PEP) is known as an inhibitor of prolidase-the enzyme that plays important role in collagen biosynthesis, the mechanism of pyruvate interconversion was considered as a regulatory switch in collagen biosynthesis. In fact, 3-MPA, specific inhibitor of phosphoenolpyruvate carboxykinase (PEPCK), contributed to up-regulation of prolidase activity, suggesting that down-regulation of PEP formation is an underlying mechanism. Since collagen biosynthesis and prolidase activity are regulated by signal induced by activated α2β1 integrin receptor as well as insulin-like growth factor-I receptor (IGF-IR), the expression of these receptors was measured by Western immunoblot analysis. The exposure of the cells to OXY contributed to decrease in IGF-IR, α2β1 integrin receptor, pERK1/2, and NF-κB p65 expressions. It was accompanied by increase in total ERK1/2 expression and induction of phosphorylation of Akt protein. The data suggest that OXY-dependent increase of collagen biosynthesis in cultured human skin fibroblasts results from activation of prolidase activity and level, induction in pAkt expression and down-regulation of pERK1/2 and NF-κB p65, the known inhibitor of collagen gene expression.
authors: Department of Medicinal Chemistry, Medical University of Bialystok, Mickiewicza 2 D, 15-222, Bialystok, Poland.
authors: Lukasz
authors: L
authors: Szoka
authors: Ewa
authors: E
authors: Karna
authors: Jerzy
authors: J
authors: Palka
mesh_terms: D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D004150:Dipeptidases / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D010119:Oxythiamine / Q000494:pharmacology*; D012867:Skin / Q000166:cytology
chemical_list: D010119:Oxythiamine; D003094:Collagen; D004150:Dipeptidases; C022449:proline dipeptidase
references: Cancer Res. 1997 Oct 1;57(19):4242-8
references: 9331084
references: Cancer Lett. 1998 May 15;127(1-2):63-70
references: 9619859
references: Mol Cell Biochem. 1998 Dec;189(1-2):177-83
references: 9879669
references: J Immunol. 1999 Apr 1;162(7):4226-34
references: 10201951
references: FEBS Lett. 1999 Jul 30;456(1):113-8
references: 10452541
references: J Biol Chem. 1990 Nov 15;265(32):19600-5
references: 2246245
references: Biochemistry. 1991 Apr 30;30(17):4160-4
references: 2021607
references: Crit Rev Biochem Mol Biol. 1993;28(1):31-81
references: 8444042
references: J Nutr. 1993 May;123(5):909-14
references: 8487102
references: J Cell Biol. 1999 Oct 18;147(2):401-16
references: 10525544
references: J Biol Chem. 1951 Nov;193(1):265-75
references: 14907713
references: Expert Opin Ther Targets. 2005 Aug;9(4):753-68
references: 16083341
references: Can J Microbiol. 2005 Oct;51(10):833-9
references: 16333342
references: J Biochem. 2007 Feb;141(2):287-92
references: 17169973
references: Mol Cell Biochem. 2008 Aug;315(1-2):61-7
references: 18491034
references: Amino Acids. 2008 Nov;35(4):731-8
references: 18320291
references: J Proteome Res. 2010 Feb 5;9(2):980-9
references: 20035555
references: Postepy Hig Med Dosw (Online). 2011;65:447-69
references: 21734329
references: DNA Cell Biol. 1999 Oct;18(10):751-61
references: 10541434
references: Comp Biochem Physiol A Mol Integr Physiol. 2000 Feb;125(2):265-71
references: 10825698
references: Exp Toxicol Pathol. 2000 May;52(2):149-55
references: 10965990
references: J Endocrinol. 2002 Jul;174(1):63-70
references: 12098664
references: J Basic Microbiol. 2003;43(6):522-9
references: 14625902
references: Am J Med. 1968 Jul;45(1):152-9
references: 4968882
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: Oncology. 1976;33(5-6):250-2
references: 1026857
references: Clin Chim Acta. 1982 Oct 27;125(2):193-205
references: 7139961
references: Life Sci. 1984 May 21;34(21):1985-98
references: 6727550
references: Clin Chim Acta. 1985 Jul 30;150(1):1-9
references: 4042361
references: Cancer Res. 1987 Feb 15;47(4):936-42
references: 3802100
references: Br J Exp Pathol. 1987 Feb;68(1):7-13
references: 3814502
references: Lab Invest. 1987 Oct;57(4):392-6
references: 2823005
references: Endocrinology. 1989 Feb;124(2):964-70
references: 2463909
references: Clin Physiol Biochem. 1989;7(3-4):128-36
references: 2805564
references: Arch Biochem Biophys. 1990 Jan;276(1):85-93
references: 2297232
references: Can Med Assoc J. 1975 Oct 18;113(8):759, 762-3
references: 803128
references: Biochim Biophys Acta. 1990 Jun 21;1049(2):171-6
references: 2364107
references: Mol Cell Biochem. 1997 Mar;168(1-2):177-83
references: 9062907
references: Pol J Pharmacol. 1996 Nov-Dec;48(6):609-13
references: 9112701
references: Tokai J Exp Clin Med. 1996 Dec;21(4-6):207-13
references: 9300982
references: J Cell Biochem. 1997 Nov 1;67(2):166-75
references: 9328822