Korean J Nutr KJN Korean Journal of Nutrition 0367-6463 2005-7121 The Korean Nutrition Society 10.4163/kjn.2009.42.6.505 Original Article Effect of Vitamin C, Silicon and Iron on Collagen Synthesis and Break-Down Enzyme Expression in the Human Dermal Fibroblast Cell (HS27) Kim Jeong Eun Lee Jinah Kim Hyunae Kim Jungmin Cho Yunhi Department of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University, Yongin 449-701, Korea. To whom correspondence should be addressed. (choyunhi@khu.ac.kr) 09 2009 30 09 2009 42 6 505 515 06 07 2009 26 08 2009 04 09 2009 © 2009 The Korean Nutrition Society 2009

Collagen is the major matrix protein in dermis and consists of proline and lysine, which are hydroxylated by prolyl hydroxylase (PH) and lysyl hydroxylase (LH) with cofactors such as vitamin C, oxygen, iron (Fe2+), ketoglutarate and silicon. The collagen degradation is regulated by matrix metalloproteinase-1 (MMP-1), of which is the major collagen-degrading proteinase whereas tissue inhibitors of metalloproteinase-1 (TIMP-1) bind to MMP-1 thereby inhibiting MMP-1 activity. In this study, we investigated the effects of vitamin C, silicon and iron on mRNA, protein expressions of PH, LH, MMP-1 and TIMP-1. The physiological concentrations of vitamin C (0-100 µM), silicon (0-50 µM) and iron (Fe2+:0-50 µM) were treated to human dermal fibroblast cells (HS27 cells) for 3 or 5days. The expression level of mRNA and protein was increased in not only PH but also LH when cells were incubated with vitamin C. A similar increase in LH mRNA or protein expression occurred when cells were incubated with silicon. Our results suggest that treatment of vitamin C and silicon increased mRNA and protein expression of PH and LH in human dermal fibroblast.

 vitamin C silicon iron prolyl hydroxylase lysyl hydroxylase

This research was supported by grants from Korea research foundation (2006-KRF-531-C00063).

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Effects of ascorbic acid, silicon and iron on prolyl hydroxylase (PH) mRNA expressions of cultured human dermal fibroblasts. Human dermal fibroblast cultures were treated with ascorbic acid (0, 5, 10, 50, 100 µM), silicon (0, 5, 10, 25, 50 µM) and Fe (0, 5, 10, 20, 30, 50, 100 µM) for 3 days (A) and 5 days (B). The expression levels of PH mRNA was analyzed by RT-PCR. The signal intensity were quantified the integrated area were percentized to the signal observed in control cells (% control). Results shown are means ± SEM (n = 3)(p < 0.05).

Effects of ascorbic acid, silicon and iron on lysyl hydroxylase (LH) mRNA expressions of cultured human dermal fibroblasts. Human dermal fibroblast cultures were treated with ascorbic acid (0, 5, 10, 50, 100 µM), silicon (0, 5, 10, 25, 50 µM) and Fe (0, 5, 10, 20, 30, 50, 100 µM) for 3 days (A) and 5 days (B). The expression levels of LH mRNA was analyzed by RT-PCR. The signal intensity were quantified the integrated area were percentized to the signal observed in control cells (% control). Results shown are means ± SEM (n = 3)(p < 0.05).

Effects of ascorbic acid, silicon and iron on prolyl hydroxylase (PH) protein expressions of cultured human dermal fibroblasts. Human dermal fibroblast cultures were treated with ascorbic acid (0, 5, 10, 50, 100 µM), silicon (0, 5, 10, 25, 50 µM) and Fe (0, 5, 10, 20, 30, 50, 100 µM) for 3 days (A) and 5 days (B). The expression levels of PH proteins was analyzed by western blotting. The signal intensity were quantified the integrated area were percentized to the signal observed in control cells (% control). Results shown are means ± SEM (n = 3)(p < 0.05).

Effects of ascorbic acid, silicon and iron on Lysyl hydroxylase (LH) protein expressions of cultured human dermal fibroblasts. Human dermal fibroblast cultures were treated with ascorbic acid (0, 5, 10, 50, 100 µM), silicon (0, 5, 10, 25, 50 µM) and Fe (0, 5, 10, 20, 30, 50, 100 µM) for 3 days (A) and 5 days (B). The expression levels of LH proteins was analyzed by western blotting. The signal intensity were quantified the integrated area were percentized to the signal observed in control cells (% control). Results shown are means ± SEM (n = 3)(p < 0.05).

Effects of ascorbic acid, silicon and iron on MMP-1 and TIMP-1 expressions of cultured human dermal fibroblasts. Human dermal fibroblast cultures were treated with ascorbic acid (0, 5, 10, 50, 100 µM), silicon (0, 5, 10, 25, 50 µM) and Fe (0, 5, 10, 20, 30, 50 µM) for 3 days. A: The expressions of MMP-1 protein was analysis by western blotting. B: The expressions of TIMP-1 was analyzed by western blotting. Results shown are means ± SEM.

Primer sequences used in RT-PCR