• Sascha David, Ute von Jan, Philipp Kümpers, Hermann Haller, and Joon-Keun Park.
• Division of Nephrology and Hypertension, Medical School Hannover, Hannover, Germany. david.sascha@mh-hannover.de
• J. Nephrol. 2013 Mar 1; 26 (2): 385-8.

BackgroundAcute kidney injury (AKI) in sepsis accompanied by functional impairment regularly lacks histologic pathology to a similar extent. Moderately reproducible - semi-quantitative - scoring systems still remain the method of choice to quantify the extent of experimental AKI. Our aim was to establish a rather simple, fast, and reproducible staining protocol to quantify loss of brush border (BB) in septic AKI.MethodsMurine septic AKI was induced by cecal ligation and puncture (CLP) and kidneys were harvested 24 h later. Binding of Phaseolus vulgaris erythroagglutinin (PHA-E) was visualized by the ABC-GO method coupled to tetranitroblue tetrazolium (TNBT). The mean BB area of five randomly chosen, non-overlapping cortical fields was analyzed by customized planimetric software.ResultsWe detected statistically significant differences in the BB/noBB pixel ratio between septic AKI, sham-operated animals, and healthy mice (CLP: 0.091 ± 0.019; SHAM: 0.141 ± 0.048, P<.001; HEALTHY controls: 0.266 ± 0.14 pixel ratio). Moreover, this ratio correlates with functional parameters of renal function (creatinine r=-0.37, BUN r=-0.54, P<.0001).ConclusionsOur results demonstrate that PHA-E lectin staining followed by planimetric quantification is a reliable tool to analyze loss of BB during experimental AKI.

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