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小鼠急性肾损伤及损伤后纤维化的缺血再灌注模型

Ischemia-reperfusion model of acute kidney injury and post injury fibrosis in mice.

作者信息

Skrypnyk Nataliya I, Harris Raymond C, de Caestecker Mark P

机构信息

Division of Nephrology, Vanderbilt University Medical Center.

出版信息

J Vis Exp. 2013 Aug 9(78):50495. doi: 10.3791/50495.

Abstract

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

摘要

缺血再灌注诱导的急性肾损伤(IR-AKI)在小鼠中被广泛用作急性肾损伤的模型,但结果往往差异很大,死亡率很高且常常未报告,这可能会混淆分析结果。双侧肾蒂夹闭常用于诱导IR-AKI,但有效夹闭压力之间的差异和/或肾脏对缺血的反应差异往往导致结果更具可变性。此外,已知较短的夹闭时间会导致更具可变性的肾小管损伤,而夹闭时间较长的双侧损伤小鼠会出现更一致的肾小管损伤,但它们通常会在损伤后的头3天内由于严重肾功能不全而死亡。为了提高损伤后的存活率并获得更一致和可预测的结果,我们开发了两种单侧缺血再灌注损伤模型,随后进行对侧肾切除术。两种手术均采用背部入路,减少了常用于小鼠IR-AKI手术的腹部剖腹术所产生的手术应激。为了诱导中度损伤,BALB/c小鼠单侧夹闭肾蒂26分钟,并同时进行对侧肾切除术。采用这种方法,50-60%的小鼠在损伤后24小时出现中度急性肾损伤,但90-100%的小鼠存活。为了诱导更严重的急性肾损伤,BALB/c小鼠肾蒂夹闭30分钟,然后在损伤后8天进行对侧肾切除术。这允许在90-100%存活的情况下对损伤后的肾脏恢复进行功能评估。使用该模型,损伤后早期的肾小管损伤以及损伤后的纤维化高度一致。

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