Urinary proteomics before and after extracorporeal circulation in patients with and without acute kidney injury

Objective

Acute kidney injury is a well-known complication with high morbidity and mortality after cardiopulmonary bypass. Cardiopulmonary bypass–associated acute kidney injury is still poorly understood.

Methods

Thirty-six patients undergoing elective cardiopulmonary bypass were enrolled. Spot urine samples before and after cardiopulmonary bypass were collected. Acute kidney injury was defined according to the RIFLE classification. To identify differentially regulated proteins after cardiopulmonary bypass, we first analyzed the urinary proteome before and after cardiopulmonary bypass. To identify differentially regulated proteins in acute kidney injury, we next compared the urinary proteome obtained on the first postoperative day between patients with and without acute kidney injury. Difference fluorescence gel electrophoresis was used to compare protein profiles and mass spectrometry to identify individual proteins.

Results

After cardiopulmonary bypass, inflammation-associated (zinc-alpha-2-glycoprotein, leucine-rich alpha-2-glycoprotein, mannan-binding lectin serine protease 2, basement membrane-specific heparan sulfate proteoglycan, and immunoglobulin kappa) or tubular dysfunction-associated (retinol-binding protein, adrenomedullin-binding protein, and uromodulin) proteins were differentially regulated. Acute kidney injury developed in 6 of 36 patients. A modified urinary albumin was increased, and zinc-alpha-2-glycoprotein and a fragment of adrenomedullin-binding protein were decreased in patients with acute kidney injury. Decreased excretion of zinc-alpha-2-glycoprotein in patients with acute kidney injury was confirmed by Western blot and enzyme-linked immunosorbent assay in an independent cohort of 22 patients with and 46 patients without acute kidney injury.

Conclusion

Cardiopulmonary bypass leads to increased urinary excretion of inflammatory proteins and markers of tubular injury. Zinc-alpha-2-glycoprotein is a potentially useful predictive marker for acute kidney injury after cardiopulmonary bypass surgery.

CTSNet classification: 25, 29, 38

Abbreviations and Acronyms: 2D, 2-dimensional, AKI, acute kidney injury, AMBP, adrenomedullin-binding protein, AUC, area under the curve, CPB, cardiopulmonary bypass, DIGE, difference gel electrophoresis, eGFR, estimated glomerular filtration rate, ELISA, enzyme-linked immunosorbent assay, GFR, glomerular filtration rate, HSPG, heparan sulfate proteoglycan, MALDI-MS, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, MALDI-MS/MS, matrix-assisted laser desorption ionization time-of-flight mass spectrometry/mass spectrometry, NGAL, neutrophil gelatinase-associated lipocalin, RBP, retinol-binding protein, SELDI-TOF-MS, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, SIRS, systemic inflammatory response syndrome, ZAG, zinc-alpha-2-glycoprotein