Department of Food Chemistry and Toxicology, Berlin Institute of Technology, Gustav-Meyer-Allee 25, D-13355 Berlin, Germany.
Metallomics. 2018 Jul 18;10(7):979-991. doi: 10.1039/c8mt00064f.
The molecular mechanisms of intestinal zinc resorption and its regulation are still topics of ongoing research. To this end, the application of suitable in vitro intestinal models, optimized with regard to their cellular composition and medium constituents, is of crucial importance. As one vital aspect, the impact of cell culture media or buffer compounds, respectively, on the speciation and cellular availability of zinc has to be considered when investigating zinc resorption. Thus, the present study aims to investigate the impact of serum, and in particular its main constituent serum albumin, on zinc uptake and toxicity in the intestinal cell line Caco-2. Furthermore, the impact of serum albumin on zinc resorption is analyzed using a co-culture of Caco-2 cells and the mucin-producing goblet cell line HT-29-MTX. Apically added albumin significantly impaired zinc uptake into enterocytes and buffered its cytotoxicity. Yet, undigested albumin does not occur in the intestinal lumen in vivo and impairment of zinc uptake was abrogated by digestion of albumin. Interestingly, zinc uptake, as well as gene expression studies of mt1a and selected intestinal zinc transporters after zinc incubation for 24 h, did not show significant differences between 0 and 10% serum. Importantly, the basolateral application of serum in a transport study significantly enhanced fractional apical zinc resorption, suggesting that the occurrence of a zinc acceptor in the plasma considerably affects intestinal zinc resorption. This study demonstrates that the apical and basolateral medium composition is crucial when investigating zinc, particularly its intestinal resorption, using in vitro cell culture.
肠道锌吸收的分子机制及其调节仍是正在研究的课题。为此,应用合适的体外肠道模型,优化其细胞组成和培养基成分至关重要。作为一个重要方面,在研究锌吸收时,必须考虑细胞培养基或缓冲化合物分别对锌的形态和细胞可用性的影响。因此,本研究旨在研究血清,特别是其主要成分血清白蛋白,对肠道细胞系 Caco-2 中锌摄取和毒性的影响。此外,还使用 Caco-2 细胞和产生粘蛋白的 goblet 细胞系 HT-29-MTX 的共培养物分析了血清白蛋白对锌吸收的影响。细胞顶端添加的白蛋白显著损害了肠细胞对锌的摄取,并缓冲了其细胞毒性。然而,体内肠腔中不会出现未消化的白蛋白,并且白蛋白的消化消除了对锌摄取的损害。有趣的是,锌摄取以及锌孵育 24 小时后 mt1a 和选定的肠道锌转运体的基因表达研究显示,0%和 10%血清之间没有显著差异。重要的是,在转运研究中,血清的基底外侧应用显著增强了锌的腔侧分数吸收,这表明血浆中锌受体的存在会显著影响肠道锌的吸收。本研究表明,在使用体外细胞培养研究锌,特别是其肠道吸收时,细胞顶端和基底外侧培养基的组成至关重要。