Cold-induced injury to porcine corneal endothelial cells and its mediation by chelatable iron - Implications for corneal preservation
Cold-induced injury to porcine corneal endothelial cells and its mediation by chelatable iron - Implications for corneal preservation
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DOI：
10.1097/01.ico.0000167880.96439.c6
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发表时间：
2006-01-01
期刊：
影响因子：
2.8
通讯作者：
de Groot, H
中科院分区：
文献类型：
--
作者：
Rauen, U;Kerkweg, U;de Groot, H
目的：在角膜的低温储存过程中，角膜内皮损伤限制了储存时间。我们以前报道了一个新的，铁依赖性的机制冷诱导损伤培养的肝细胞。在这项研究中，我们试图评估是否角膜内皮细胞招致类似的injure.Methods：培养的猪角膜内皮细胞暴露于4 ℃的细胞培养基，Krebs-Henseleit缓冲液，Optisol-GS解决方案，或McCarey-Kaufman培养基5小时至14天，然后在细胞培养条件下（3小时）复温。培养物进行了评估的致死性细胞损伤（LDH释放）;细胞，核，和线粒体形态学的改变;脂质过氧化;和线粒体membrane potential.Results：角膜内皮细胞持续实质性损伤后冷藏和复温的细胞培养基（47% +/- 8%和64% +/- 20%细胞死亡后2和5天冷藏，分别）。损伤显示出一些凋亡特征，并且细胞在细胞死亡发生之前丧失线粒体膜电位。铁螯合剂去铁胺、1，10-菲咯啉和2，2 '-联吡啶以及抗氧化剂丁基化羟基甲苯完全抑制了这种细胞损伤。显着的铁依赖性细胞损伤和脂质过氧化反应也发生在冷孵育过程中和之后，在Krebs-Henseleit缓冲液，最重要的是，铁依赖性细胞损伤也观察到冷孵育后的Optisol解决方案，并在McCarey-Kaufman medium.Conclusions：培养的猪角膜内皮细胞产生强烈的铁依赖性损伤引起的低温。这种冷诱导的损伤可能为已知的角膜内皮对低温保存损伤的易感性提供了解释，因此可能适用于治疗干预（即，通过铁螯合剂）。
Purpose: During hypothermic storage of the cornea, corneal endothelial damage restricts storage times. We previously reported a new, iron-dependent mechanism of cold-induced injury to cultured liver cells. In this study, we sought to evaluate whether corneal endothelial cells incur a similar kind of injury.Methods: Cultured porcine corneal endothelial cells were exposed to 4 degrees C in either cell culture medium, Krebs-Henseleit buffer, Optisol-GS solution, or McCarey-Kaufman medium for 5 hours to 14 days and then rewarmed under cell culture conditions (3 hours). The cultures were assessed for lethal cell injury (LDH release); cellular, nuclear, and mitochondrial morphologic alterations; lipid peroxidation; and mitochondrial membrane potential.Results: Corneal endothelial cells sustained substantial injury following cold storage and rewarming in cell culture medium (47% +/- 8% and 64% +/- 20% cell death after 2 and 5 days cold storage, respectively). The injury displayed some apoptotic features, and cells lost mitochondrial membrane potential before cell death occurred. The iron chelators deferoxamine, 1,10-phenanthroline, and 2,2'-dipyridyl and the antioxidant butylated hydroxytoluene completely inhibited this cell injury. Marked iron-dependent cell injury and lipid peroxidation also occurred during and after cold incubation in Krebs-Henseleit buffer and, most importantly, iron-dependent cell injury was also observed after cold incubation in Optisol solution and in McCarey-Kaufman medium.Conclusions: Cultured porcine corneal endothelial cells incur a strong iron-dependent injury elicited by hypothermia. This cold-induced injury might provide an explanation for the known corneal endothelial susceptibility to hypothermic preservation injury, which thus might be amenable to therapeutic interventions (ie, by iron chelators).