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血红素载体蛋白1(HCP1):在培养细胞中的表达及功能研究

Haem carrier protein 1 (HCP1): Expression and functional studies in cultured cells.

作者信息

Latunde-Dada Gladys O, Takeuchi Ken, Simpson Robert J, McKie Andrew T

机构信息

Department of Biochemistry, School of Biomedical and Health Sciences, Franklin Wilkins Building, King's College London, 150 Stamford Street, London, SE1 9NH, United Kingdom.

出版信息

FEBS Lett. 2006 Dec 22;580(30):6865-70. doi: 10.1016/j.febslet.2006.11.048. Epub 2006 Nov 29.

Abstract

Haem released from digestion and breakdown of meat products provides an important source of dietary iron, which is readily absorbed in the proximal intestine. The recent cloning and characterization of a haem carrier protein 1 (HCP 1) has provided a candidate intestinal haem transporter. The current studies describe the expression and functional analysis of HCP1 in cultured Caco-2 cells, a commonly used model of human intestinal cells. HCP1 mRNA expression in other cell types was also studied. The uptake of (55)Fe labeled haem was determined in cells under different experimental conditions and HCP1 expression was measured by RT-PCR and immunohistochemistry. mRNA and protein expressions increased in Caco-2 cells transduced with HCP1 adenoviral plasmid, and consequently (55)Fe haem uptake was higher in these cells. Haem uptake was also increased in fully differentiated Caco-2 cells compared to undifferentiated cells. Preincubation of cells with desferrioxamine (DFO, to deplete cells of iron) had no effect on HCP1 expression or haem uptake. Treatment with CdCl(2) (to induce haem oxygenase, HO-1) enhanced HCP1 expression and increased haem uptake into the cells. HCP1 expression and function were found to be adaptive to the rate of haem degradation by HO-1. Furthermore, HCP1 expression in different cells implies a functional role in tissues other than the duodenum.

摘要

肉类产品消化和分解过程中释放的血红素提供了膳食铁的重要来源,这种铁易于在近端肠道被吸收。最近克隆和鉴定的血红素载体蛋白1(HCP 1)为肠道血红素转运体提供了一个候选蛋白。当前的研究描述了HCP1在常用的人肠道细胞模型——培养的Caco-2细胞中的表达及功能分析。还研究了HCP1 mRNA在其他细胞类型中的表达。在不同实验条件下测定细胞对(55)Fe标记血红素的摄取,并通过RT-PCR和免疫组织化学检测HCP1的表达。用HCP1腺病毒质粒转导的Caco-2细胞中,mRNA和蛋白表达增加,因此这些细胞中(55)Fe血红素摄取更高。与未分化细胞相比,完全分化的Caco-2细胞中的血红素摄取也增加。用去铁胺(DFO,使细胞耗尽铁)预孵育细胞对HCP1表达或血红素摄取没有影响。用CdCl2处理(诱导血红素加氧酶,HO-1)可增强HCP1表达并增加细胞对血红素的摄取。发现HCP1的表达和功能可适应HO-1介导的血红素降解速率。此外,HCP1在不同细胞中的表达意味着它在十二指肠以外的组织中也发挥功能作用。

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