以Caco-2細胞模式初探Hepcidin對小腸細胞鐵調節蛋白活性之影響

李怡慧

doi:10.6342/NTU.2005.02621

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以Caco-2細胞模式初探Hepcidin對小腸細胞鐵調節蛋白活性之影響

Effect of Hepcidin on Iron regulatory Proteins Binding Activity in Caco-2 cells

李怡慧 , Masters　　Advisor：蕭寧馨　　

繁體中文 DOI： 10.6342/NTU.2005.02621

Iron regulatory proteins ； Hepcicin ； Caco-2 cell ； iron regulatory proteins ； Hepcicin ； Caco-2 cells

Abstract │ Reference (99) │ Altmetrics

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In mammals, iron balance is maintained by meticulous regulation of iron absorption from the duodenum, which responds to body iron stores and erythropoietic iron demand. Hepcidin (Hepc) is postulated to be a circulating peptide specifically produced in the liver and its expression is regulated by iron storage, inflammation, and anemia. It has been suggested that Hepc could act on the small intestine and regulate the iron absorption. Iron regulatory proteins (IRPs) are crucial components to regulated cellular iron homeostasis. We adopted an in vitro Caco-2 cell model to assess the effect of hepcidin on iron regulatory proteins activity. Hepc20 and Hepc25, composed of 20 and 25 amino acid residues, respectively were synthesized by solid phase peptide synthesis. The primary-structured products were purified and air-oxidized in the presence of the cysteine/cystine redox phosphate buffer at pH 7.4 to form the biologically active molecules containing four disulfide bonds. The molecule weight of primary-structured Hepc20 and Hepc25 were 2,199.9 and 2,797.2; the molecule weight of tertiary -structured Hepc20 and Hepc25 were 2,191.1 and 2,787.9. These tertiary-structured hepcidin were used for cell treatment. Caco-2 cells were exposed to Hepc20 and Hepc25 in the basolateral site prior to the measurement of IRPs activity. Incubation with Hepc20 and Hepc25 at 10 μM for 12 and 24 hours has no effect on IRPs activity. As concentration of treatment increased, the activity of IRPs was still unchanged. IRPs activity was also unaffected by Hepc25 purchased from the market. These results showed that Hepc20 and Hepc25 had no effect on IRPs activity in Caco-2 cells. These phenomena need to be further studied.

Reference ( 99 ) 〈TOP〉

李慧玲 (2004) 以Caco-2細胞模式探討hepcidin對鐵吸收相關分子表現之影響。台大微生物與生化所碩士論文。
連結：
Aisen, P., Wessling-Resnick, M., Leibold, E. A. (1999) Iron metabolism. Curr Opin Chem Biol. 3: 200-206.
連結：
Anderson, G. J., Frazer, D. M., Wilkins, S. J., Becker, E. M., Millard, K. N., Murphy, T. L., McKie, A. T.＆Vulpe, C. D. (2002) Relationship between intestinal iron-transporter expression, hepatic hepcidin levels and the control of iron absorption. Biochem Soc Trans. 30: 724-726.
連結：
Andrews, N. C. (2004) Anemia of inflammation: the cytokine-hepcidin link. J Clin Invest. 113: 1251-1253.
連結：
Anfinsen, C. B. (1973) Principles that govern the folding of protein chains. Science. 181: 223-230.
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