Carbamylated erythropoietin-FC fusion protein and recombinant human erythropoietin during porcine kidney ischemia/reperfusion injury
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To test the hypothesis that a carbamylated EPO-FC fusion protein (cEPO-FC) or recombinant human erythropoietin (rhEPO) would protect against kidney ischemia/reperfusion (I/R) injury in pigs with atherosclerosis.
Anesthetized and mechanically ventilated animals received cEPO-FC (50 μg kg−1), rhEPO (5,000 IU kg−1), or vehicle (n = 9 per group) prior to 120 min of aortic occlusion and over 4 h of reperfusion. During aortic occlusion, mean arterial pressure (MAP) was maintained at 80–120 % of baseline values by esmolol, nitroglycerin, and ATP. During reperfusion, noradrenaline was titrated to keep MAP at pre-ischemic levels. Blood creatinine and neutrophil gelatinase-associated lipocalin (NGAL) levels, creatinine clearance, fractional Na+ excretion, and HE and PAS staining were used to assess kidney function and histological damage. Plasma interleukin-6, tumor necrosis factor-α, nitrate + nitrite and 8-isoprostane levels were measured to assess systemic inflammation, and nitrosative and oxidative stress.
I/R caused acute kidney injury with reduced creatinine clearance, increased fractional Na+ excretion and NGAL levels, moderate to severe glomerular and tubular damage and apoptosis, systemic inflammation and oxidative and nitrosative stress, but there were no differences between the treatment groups. Pre-ischemia nitrate + nitrite and 8-isoprostanes levels were lower and higher, respectively, than in healthy animals of a previous study, and immune histochemistry showed higher endothelial nitric oxide synthase and lower EPO receptor expression in pre-ischemia kidney biopsies than in biopsies from healthy animals.
In swine with atherosclerosis, rhEPO and cEPO-FC failed to attenuate prolonged ischemia-induced kidney injury within an 8-h reperfusion period, possibly due to reduced EPO receptor expression resulting from pre-existing oxidative stress and/or reduced NO release.
KeywordsGlomerular filtration Tubular reabsorption Creatinine clearance Fractional Na+ sodium excretion Neutrophil gelatinase-associated lipocalin HE staining PAS staining Apoptosis Cytokines 8-isoprostanes Nitric oxide Endothelial NO synthase EPO receptor
The carbamylated EPO-FC fusion protein (cEPO-FC) and the recombinant human EPO (rhEPO) were kindly provided by Polymun Scientific GmbH, Klosterneuburg, Austria. We are indebted to Tanja Schulz, Rosemarie Mayer, and Ingrid Eble for skillful technical assistance.
Conflicts of interest
Brigitta Vcelar is responsible for preclinical research at and is an employee of Polymun Scientific GmbH (Klosterneuburg, Austria), a company involved in the commercial development of cEPO-FC, but holds no equity in that company nor related to the molecules investigated. The other authors declare that they have no competing interests at all.
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