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采用 Caco-2 细胞体外通量和生物转化研究及 ICP-MS 检测法评估无机和有机硒化合物的肠道吸收。

Estimating intestinal absorption of inorganic and organic selenium compounds by in vitro flux and biotransformation studies in Caco-2 cells and ICP-MS detection.

机构信息

Department of Pharmaceutics and Analytical Chemistry, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, Denmark.

出版信息

Biol Trace Elem Res. 2012 Feb;145(2):248-56. doi: 10.1007/s12011-011-9174-y. Epub 2011 Aug 24.

Abstract

The aim of the present work was to compare and estimate absorption and biotransformation of selected selenium compounds by studying their fluxes across Caco-2 cells. Five different selenium compounds, selenomethionine (SeMet), Se-methylselenocysteine (MeSeCys), selenate, selenite, and methylseleninic acid (MeSeA), were applied to Caco-2 cells in a concentration of 10 μM, and fluxes in both directions were studied for 2 h. Fluxes of selenite and MeSeA in the presence of excess reduced glutathione (selenite + GSH and MeSeA + GSH) and flux of MeSeA in the presence of excess cysteine (MeSeA + Cys) were also studied. Selenium absorptive and exsorptive fluxes and accumulation in cell cytosol were analyzed by means of flow injection inductively coupled plasma mass spectrometry (ICP-MS). Absorptive flux of SeMet, MeSeCys, and selenate showed values correlating to complete in vivo absorption, while selenite and MeSeA fluxes correlated to poor in vivo absorption. Speciation analysis of cell lysate and donor and receptor solutions by LC-ICP-MS showed limited transformation of all selenium compounds. Extensive transformation as well as significantly increased absorptive flux was observed when co-administering selenite with glutathione compared to administering selenite alone. These observations are possibly due to formation of selenodiglutathione (GS-Se-SG) which may be absorbed differently than selenite. Concomitant application of GSH or cysteine with MeSeA resulted in extensive transformation of MeSeA, including volatile species, whereas no significant increases in fluxes were observed. In summary, the absorption of selenite selenate and the selenoamino acids is considered complete under physiological conditions, but the absorption mechanisms and metabolism of the compounds are different.

摘要

本工作旨在通过研究它们穿过 Caco-2 细胞的通量来比较和估计选定硒化合物的吸收和生物转化。将五种不同的硒化合物,即硒代蛋氨酸(SeMet)、硒代半胱氨酸(MeSeCys)、硒酸盐、亚硒酸盐和甲基亚硒酸(MeSeA),以 10 μM 的浓度应用于 Caco-2 细胞,并在 2 小时内研究两个方向的通量。还研究了在过量还原型谷胱甘肽存在下(亚硒酸盐+GSH 和 MeSeA+GSH)的亚硒酸盐和 MeSeA 的吸收通量以及在过量半胱氨酸存在下的 MeSeA 的通量(MeSeA+Cys)。通过流动注射电感耦合等离子体质谱(ICP-MS)分析细胞胞质溶胶中的硒吸收和外排通量和积累。SeMet、MeSeCys 和硒酸盐的吸收通量值与完全的体内吸收相关,而亚硒酸盐和 MeSeA 的通量与体内吸收不良相关。通过 LC-ICP-MS 对细胞裂解物以及供体和受体溶液进行的形态分析表明,所有硒化合物的转化都受到限制。与单独给予亚硒酸盐相比,当与谷胱甘肽共同给予亚硒酸盐时,观察到广泛的转化以及吸收通量的显著增加。这些观察结果可能是由于形成了硒代二谷胱甘肽(GS-Se-SG),其可能与亚硒酸盐的吸收方式不同。同时给予 GSH 或半胱氨酸与 MeSeA 导致 MeSeA 的广泛转化,包括挥发性物质,而没有观察到通量的显著增加。总之,在生理条件下,亚硒酸盐、硒酸盐和硒代氨基酸的吸收被认为是完全的,但化合物的吸收机制和代谢是不同的。

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