Sauvant Christoph, Schneider Reinhard, Holzinger Hildegard, Renker Sylvia, Wanner Christoph, Gekle Michael
Physiologisches Institut, Bay. Julius-Maximilians Universitat Wurzburg, Germany.
Cell Physiol Biochem. 2009;24(5-6):567-76. doi: 10.1159/000257513. Epub 2009 Nov 4.
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 (alphaSMA), 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.
缺血性急性肾损伤（iAKI）是器官移植中的常见事件，在严重手术期间也可能发生。为了深入了解缺血诱导的近端小管细胞水平的改变机制，我们使用大鼠近端小管细胞系NRK-52E建立了一个缺血再灌注体外模型。在这个特定模型中，我们使用低容量缓冲系统和缺氧箱，将酸中毒、缺氧和无糖血症同时应用2小时。此后，通过在标准条件下将细胞培养长达48小时来模拟再灌注。为了验证该系统，我们研究了是否能观察到在现有的缺血再灌注体内模型中发生的效应。即坏死、凋亡和缺血再灌注诱导蛋白（IRIP）的诱导、去分化（αSMA）、炎症（MCP-1）、诱导型一氧化氮合酶（iNOS）、PGE(2)的释放以及有机阴离子的基底外侧摄取。事实上，所有参数在缺血后再灌注期间均如体内情况所描述的那样出现变化。综上所述，我们建立了一个缺血后近端小管细胞再灌注损伤的体外模型，显示出体内描述的典型变化。此外，我们的模型系统适用于单独应用与缺血相关的典型损伤（如单独的酸中毒、单独的缺氧、单独的无糖血症），以便在细胞水平上更深入了解导致肾脏再灌注损伤的机制事件。
