Protective effects of ezrin on cold storage preservation injury in the pig kidney proximal tubular epithelial cell line (LLC-PK1).
Protective effects of ezrin on cold storage preservation injury in the pig kidney proximal tubular epithelial cell line (LLC-PK1).
复制标题
埃兹蛋白对猪肾近端肾小管上皮细胞系（LLC-PK1）冷藏保存损伤的保护作用。
DOI：
10.1097/tp.0b013e3181a43f18
复制
发表时间：
2009
期刊：
影响因子：
6.2
通讯作者：
Mangino,MartinJ
中科院分区：
文献类型：
--
作者：
Tian,Tao;Lindell,SusanneL;Henderson,ScottC;Mangino,MartinJ
背景：冷保存和热再灌注引起的肾损伤已被充分证实，包括组织水肿、细胞肿胀、ATP耗竭、钙毒性和氧化应激。然而，更常见的近端机制尚未确定，这可能会限制有效的临床治疗策略的发展。以前的工作表明，许多细胞骨架结构的影响，冷保存和再灌注，包括膜丰富ezrin相关的复合物。本研究的目的是调查是否亚板层细胞骨架蛋白ezrin是因果关系参与冷保存损伤在肾小管epithelial cells.Methods。我们创建了一个稳定转染的细胞系（LLC-EZ）使用猪肾近端肾小管上皮细胞系（LLC-PK 1），组成型过表达野生型ezrin。将这些细胞冷保存在威斯康星州大学溶液中，并在体外再灌注以模拟肾小管保存损伤，通过生化、代谢、功能和结构终点来评估。（乳酸脱氢酶释放），线粒体活性（ATP合成、脱氢酶活性和线粒体内膜电位），保护细胞膜微绒毛和线粒体结构。再灌注诱导的细胞凋亡也显着减少LLC-EZ细胞过表达ezrin.Conclusions。增强ezrin表达保护肾小管上皮细胞免受冷藏保存损伤，可能是通过膜或线粒体机制。
Background.Renal damage caused by cold preservation and warm reperfusion has been well documented and involves tissue edema, cell swelling, ATP depletion, calcium toxicity, and oxidative stress. However, more common proximal mechanisms have not been identified, which may limit the development of effective clinical treatment strategies. Previous work indicates that many cytoskeletal structures are affected by cold preservation and reperfusion, including membrane-rich ezrin-associated complexes. The aim of this study was to investigate whether the sublamellar cytoskeletal protein ezrin is causally involved in cold preservation injury in renal tubule epithelial cells.Methods.We created a stably transfected cell line (LLC-EZ) using the pig kidney proximal tubular epithelial cell line (LLC-PK1), which constitutively overexpresses wild-type ezrin. These cells were cold stored in University of Wisconsin Solution and reperfused in vitro to model renal tubule preservation injury, which was assessed by biochemical, metabolic, functional, and structural endpoints.Results.Overexpression of ezrin increased cell viability (lactate dehydrogenase release), mitochondrial activity (ATP synthesis, dehydrogenase activity, and inner mitochondrial membrane potential), and protected the structure of cell membrane microvilli and mitochondria after cold storage preservation injury. Reperfusion-induced apoptosis was also significantly reduced in LLC-EZ cells overexpressing ezrin.Conclusions.Enhanced ezrin expression protects tubule epithelial cells from cold storage preservation injury, possibly by membrane or mitochondrial mechanisms.