Key Laboratory of Animal Nutrition & Feed Science, Zhejiang Province, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Nutrients. 2019 Feb 26;11(3):485. doi: 10.3390/nu11030485.
This experiment was conducted to investigate the transport characteristics of iron from ferrous bisglycinate (Fe-Gly) in intestinal cells. The divalent metal transporter 1 (DMT1)-knockout Caco-2 cell line was developed by Crispr-Cas9, and then the cells were treated with ferrous sulfate (FeSO₄) or Fe-Gly to observe the labile iron pool and determine their iron transport. The results showed that the intracellular labile iron increased significantly with Fe-Gly or FeSO₄ treatment, and this phenomenon was evident over a wide range of time and iron concentrations in the wild-type cells, whereas in the knockout cells it increased only after processing with high concentrations of iron for a long time ( < 0.05). DMT1-knockout suppressed the synthesis of ferritin and inhibited the response of iron regulatory protein 1 (IRP-1) and IRP-2 to these two iron sources. The expression of peptide transporter 1 (PepT1) was not altered by knockout or iron treatment. Interestingly, the expression of zinc-regulated transporter (ZRT) and iron-regulated transporter (IRT)-like protein 14 (Zip14) was elevated significantly by knockout and iron treatment in wild-type cells ( < 0.05). These results indicated that iron from Fe-Gly was probably mainly transported into enterocytes via DMT1 like FeSO₄; Zip14 may play a certain role in the intestinal iron transport.
本实验旨在研究二价甘氨酸亚铁(Fe-Gly)中铁在肠道细胞中的转运特性。利用 Crispr-Cas9 构建了二价金属转运蛋白 1(DMT1)敲除的 Caco-2 细胞系,然后用硫酸亚铁(FeSO₄)或 Fe-Gly 处理细胞,观察不稳定铁池并确定其铁转运。结果表明,细胞内不稳定铁随着 Fe-Gly 或 FeSO₄处理而显著增加,这种现象在野生型细胞中在广泛的时间和铁浓度范围内都很明显,而在敲除细胞中,只有在长时间(<0.05)处理高浓度铁后才会增加。DMT1 敲除抑制了铁蛋白的合成,并抑制了铁调节蛋白 1(IRP-1)和 IRP-2 对这两种铁源的反应。肽转运蛋白 1(PepT1)的表达不受敲除或铁处理的影响。有趣的是,锌调节转运蛋白(ZRT)和铁调节转运蛋白(IRT)样蛋白 14(Zip14)的表达在野生型细胞中,无论是敲除还是铁处理,都显著上调(<0.05)。这些结果表明,Fe-Gly 中的铁可能主要通过类似于 FeSO₄ 的 DMT1 进入肠细胞;Zip14 可能在肠道铁转运中起一定作用。