Flores Sebastian E, Day Andrew S, Keenan Jacqueline I
Department of Surgery, University of Otago Christchurch, PO Box 4345, Christchurch, New Zealand.
Biometals. 2015 Feb;28(1):143-50. doi: 10.1007/s10534-014-9810-z. Epub 2014 Nov 27.
Despite the evidence suggesting a role for Helicobacter pylori in the induction of systemic iron deficiency anaemia, little is known about the possibility of infection-associated changes in cellular iron homeostasis at the gastric epithelium. In this study we compared four different techniques for measuring iron in AGS cells, a gastric epithelial cell line that is widely used to model to H. pylori infection in vitro. Inductively coupled plasma-mass spectrometry proved to be an efficient method, but only when large numbers of cells were used. Two colorimetric assays that included the use of concentrated hydrochloric acid with or without potassium ferrocyanide detected iron in the micromolar but not the nanomolar range in cell-free standards. However, the third colorimetric assay that incorporated ferrozine proved to be highly accurate at detecting iron in the nanomolar range, and was able to detect iron in AGS cells, Moreover, using this assay, we were able to show that the level of iron in H. pylori-infected AGS cells is significantly increased when compared to uninfected cells.
尽管有证据表明幽门螺杆菌在系统性缺铁性贫血的诱发中起作用，但对于胃上皮细胞中铁稳态的感染相关变化的可能性却知之甚少。在本研究中，我们比较了四种不同的测量AGS细胞中铁含量的技术，AGS细胞是一种广泛用于体外模拟幽门螺杆菌感染的胃上皮细胞系。电感耦合等离子体质谱法被证明是一种有效的方法，但仅在使用大量细胞时才有效。两种比色法，包括使用浓盐酸（有无亚铁氰化钾），在无细胞标准品中检测到的铁处于微摩尔而非纳摩尔范围。然而，第三种结合了亚铁嗪的比色法在检测纳摩尔范围内的铁时被证明非常准确，并且能够检测AGS细胞中的铁。此外，使用该方法，我们能够表明，与未感染细胞相比，幽门螺杆菌感染的AGS细胞中的铁水平显著增加。
