Quantification of experimental acute kidney injury by computer-assisted imaging of lectin phytohemagglutinin E.
BACKGROUND
Acute 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.


METHODS
Murine 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.


RESULTS
We 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).


CONCLUSIONS
Our results demonstrate that PHA-E lectin staining followed by planimetric quantification is a reliable tool to analyze loss of BB during experimental AKI.
Division of Nephrology and Hypertension, Medical School Hannover, Hannover, Germany. david.sascha@mh-hannover.de
Sascha
S
David
Ute
U
von Jan
Philipp
P
Kümpers
Hermann
H
Haller
Joon-Keun
JK
Park
D058186:Acute Kidney Injury / Q000097:blood / Q000382:microbiology / Q000473:pathology* / Q000503:physiopathology; D000818:Animals; D015415:Biomarkers / Q000097:blood; D001806:Blood Urea Nitrogen; D002432:Cecum / Q000382:microbiology / Q000601:surgery*; D004396:Coloring Agents; D003404:Creatinine / Q000097:blood; D004195:Disease Models, Animal; D007090:Image Interpretation, Computer-Assisted; D007668:Kidney / Q000382:microbiology / Q000473:pathology* / Q000503:physiopathology; D008026:Ligation; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008871:Microvilli / Q000473:pathology; D010835:Phytohemagglutinins; D011237:Predictive Value of Tests; D011677:Punctures; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods*; D013778:Tetrazolium Salts
D015415:Biomarkers; D004396:Coloring Agents; D010835:Phytohemagglutinins; D013778:Tetrazolium Salts; C067494:erythroagglutinating phytohemagglutinin; C028130:tetranitrotetrazolium blue; D003404:Creatinine