Implementation of an in vitro Model System for Investigation of Reperfusion Damage after Renal Ischemia
Implementation of an in vitro Model System for Investigation of Reperfusion Damage after Renal Ischemia
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DOI：
10.1159/000257513
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发表时间：
2009-01-01
影响因子：
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通讯作者：
Gekle, Michael
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文献类型：
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作者：
Sauvant, Christoph;Schneider, Reinhard;Gekle, Michael
缺血性急性肾损伤（iAKI）是器官移植中的常见事件，可能发生在严重手术期间。为了获得近端小管细胞水平缺血诱导的变化的机制的见解，我们建立了一个缺血和再灌注的体外模型，使用大鼠近端小管细胞系NRK-52 E。在这个特定的模型中，我们同时应用酸中毒，缺氧和血糖不高一起2小时，使用低容量缓冲系统和缺氧室。此后，通过随后在标准条件下培养细胞长达48小时来模拟再灌注。为了验证该系统，我们研究了是否可以观察到在现有的缺血和再灌注体内模型中发生的效果。即，诱导坏死、凋亡和缺血再灌注诱导蛋白（IRIP）、去分化（aSMA）、炎症（MCP-1）、诱导型NO合酶（iNOS）、PGE（2）的释放和有机阴离子的基底侧摄取。事实上，所有参数都如所描述的缺血后再灌注期间的体内情况那样发展。总之，我们建立了一个近端小管细胞缺血再灌注损伤的体外模型，显示了体内描述的典型变化。此外，我们的模型系统适合于与缺血相关的典型损伤（例如，单独的酸中毒、单独的缺氧、单独的血糖缺乏）的隔离应用，以获得对导致细胞水平上的肾脏再灌注损伤的机制事件的更多了解。版权所有（C）2009 S. Karger AG，巴塞尔
Ischemic acute kidney injury (iAKI) is a common event in organ transplantation and may occur during severe surgery. To gain mechanistic insights into ischemia-induced alterations at the level of proximal tubule cells we set up an in vitro model of ischemia and reperfusion using the rat proximal tubule cell line NRK-52E. In this particular model we simultaneously applied acidosis, hypoxia and aglycemia together for 2h, using low volume buffer systems and a hypoxia chamber. Thereafter reperfusion was mimicked by subsequently culturing the cells for up to 48h under standard conditions. In order to validate the system we investigated whether effects that take place in existing in vivo models of ischemia and reperfusion can be observed. Namely, induction of necrosis, apoptosis and of ischemia reperfusion induced protein (IRIP), dedifferentiation (aSMA), inflammation (MCP-1), inducible NO-synthase (iNOS), release of PGE(2) and basolateral uptake of organic anions. In fact, all parameters developed as described for the in vivo situation during reperfusion after ischemia. Taken altogether we have established an in vitro model of proximal tubule cell reperfusion damage after ischemia, showing typical changes described in vivo. Additionally, our model system is suitable for isolated application of the typical insults associated with ischemia (e.g. acidosis alone, hypoxia alone, aglycemia alone), in order to obtain more insight into the mechanistic events that lead to reperfusion damage in the kidney on the cellular level. Copyright (C) 2009 S. Karger AG, Basel