J Vet Sci.  2017 Dec;18(4):507-514. 10.4142/jvs.2017.18.4.507.

Quantitative evaluation of renal parenchymal perfusion using contrast-enhanced ultrasonography in renal ischemia-reperfusion injury in dogs

• Lee G
• Jeon S
• Lee SK
• Cheon B
• Moon S
• Park JG
• Cho KO
• Choi J

• KMID: 2398510
• DOI: http://doi.org/10.4142/jvs.2017.18.4.507

Abstract

This study evaluated whether renal perfusion changes can be noninvasively estimated by using contrast-enhanced ultrasonography (CEUS) in renal ischemia-reperfusion injury and investigated the correlation between renal perfusion measured by CEUS and necrosis and apoptosis of renal tubular epithelial cells. In six dogs with experimentally induced renal ischemia-reperfusion injury, changes in time to peak intensity, peak intensity, and area under the curve were measured on CEUS. Peak intensity and area under the curve of the renal cortex began to decrease on day 1 (about 20% lower than baseline) and reached the lowest levels (about 50% of baseline) on day 4. They then gradually increased until day 10, at which time peak intensity was about 87% and area under the curve was about 95% of baseline; neither fully recovered. Both parameters were strongly correlated with the necrosis scores on histopathologic examination on day 4 (r = −0.810 of peak intensity and r = −0.886 of area under the curve). CEUS allowed quantitative evaluation of perfusion changes in acute renal ischemia-reperfusion injury, and CEUS results were correlated with renal tubular damage on histopathologic examination. Thus, CEUS could be a noninvasive, quantitative diagnostic method for determining progress of renal ischemia-reperfusion injury.

Keyword

contrast-enhanced ultrasonography; dogs; renal tubular damage; reperfusion injury; time-intensity curve

MeSH Terms

Animals
Apoptosis
Dogs*
Epithelial Cells
Evaluation Studies as Topic*
Methods
Necrosis
Perfusion*
Reperfusion Injury*
Ultrasonography*

Figure

• 

  Fig. 1 Schedule diagram showing timing of laboratory examination, power Doppler (PD) and contrast-enhanced ultrasonography (CEUS), and histopathologic examination. Laboratory study included assessment of blood pressure, serum blood urea nitrogen and creatinine, urine-specific gravity and urinary protein to creatinine ratio. The number of animals used at each day is shown.

• 

  Fig. 2 Two regions of interest were placed over the renal cortex (yellow circle) and medulla (blue circle) (A), and time-intensity curves were created (B). After contrast-enhanced ultrasonography contrast agent introduction, the renal cortex showed a rapid, homogeneous, and intense enhancement pattern, while the medulla was enhanced more gradually and with a heterogeneous pattern.

• 

  Fig. 3 Contrast-enhanced ultrasonography images obtained at the time of peak intensity in the renal cortex at baseline (A), day 4 (B), and day 10 (C). At baseline, the renal cortex showed a homogeneous and intense enhancement pattern. At day 4, the renal cortex showed noticeably reduced contrast enhancement intensity. At day 10, there was increased intensity compared to that at day 4.

• 

  Fig. 4 Gross, histopathologic observations and TUNEL assays in the 4 and 10 days after ischemia-reperfusion injury to the kidneys. (A) Severe multiple linear paleness features (arrows) in the cortex of kidney sampled at 4 days after ischemia-reperfusion injury. Note the congestion of the medulla. (B) Moderate diffuse paleness in the cortex of the kidney sampled at 10 days after ischemia-reperfusion injury. (C) Representative histopathological lesions in the cortex of kidney sampled at 4 days after ischemia-reperfusion injury. Proximal convoluted tubular epithelial cells underwent necrosis characterized by the absence of nucleus (arrow) or intracellular mineralization (double arrow). Proximal and distal convoluted tubules contained detached cells, necrotic cell debris, and protein-rich fluids (asterisk). (D) Representative histopathological lesions in the cortex of kidney sampled at 10 days after ischemia-reperfusion injury. The epithelial cells in the proximal convoluted tubule were regenerated, showing attenuation of the swollen cuboidal cells with dense nuclei (double arrow). Necrotic cells lacking nuclei or with pyknosis (arrow) were present. The distal convoluted tubules contained protein-rich materials. (E) Cortex of kidney sampled at 4 days after ischemia-reperfusion injury. Apoptotic cells were detected in the distal or collecting tubules rather than in epithelial cells in the proximal tubules, which usually underwent necrosis. (F) Cortex of the kidney sampled at 10 days after ischemia-reperfusion injury. Apoptotic cells were detected in the proximal and distal tubules. H&E stain (C and D), terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling (E and F). Scale bars = 1 cm (A and B), 50 µm (C–F).

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