The Effect of Cyanate on Type I Collagen gene Expression by Dermal Fibroblasts in Culture

Park, Jae Hong; Kim, Byung Chun; Lee, Kyu Suk

Abstract

BACKGROUND
Fibrosis is a common response to various insults or injuries and can be the outcome of any perturbation in the cellular function of any tissue. Peritoneum is always exposed to the waste products during peritoneal dialysis. In aqueous solution, there is partial and spontaneous decomposition of urea to ammonia, carbonate and cyanate. Cyanate reacts irreversibly with the N-terminal groups of amino acid, peptides and many proteins by a process known as carbamylation and may contribute to peritoneal injury with fibrosis. But only little is known about the molecular and cellular mechanism underlying the effect of cyanate on the expression of type I collagen in cultured skin fibroblasts. OBJECTIVE: The purpose of this study was to examine the effect of cyanate on type I collagen gene expression in cultured skin fibroblasts. METHODS: In this study, the effects of cyanate were examined by Northern blot hybridization, chloramphenicol acetyltransferase(CAT) assay, and immunohistochemical stain in cultured human fibroblasts. RESULTS: In Northern blot hybridization, steady-state levels of alpha1(I) procollagen mRNA were increased 2-fold at 100 micromol of cyanate compared to untreated control. Cyanate caused an alteration in the alpha1(I) procollagen mRNA expression in a dose-related fashion. In CAT assay, the relative CAT activity was 1.0 in the untreated control, 0.9 at a concentration of 0.1 micromol, 2.3 at 10 micromol, and 2.6 at a 100 micromol. Cyanate caused a marked increase on type I collagen gene promotor activity. In imunohistochemical stain with anti-type I collagen antibody, type I collagen was markedly increased in cultured fibroblasts by cyanate. CONCLUSION: These results indicate that cyanate may be a powerful up-regulator of type I collagen production through the transcriptional activation of gene expression in cultured skin fibroblasts.