Influence of Vitamin C on Antioxidant Capacity of In Vitro Perfused Porcine Kidneys.
Systemic and localized ischemia and reperfusion injury remain clinically relevant issues after organ transplantation and contribute to organ dysfunctions, among which acute kidney injury is one of the most common. An in vitro test-circuit for normothermic perfusion of porcine kidneys after warm ischemia was used to investigate the antioxidant properties of vitamin C during reperfusion. Vitamin C is known to enhance microcirculation, reduce endothelial permeability, prevent apoptosis, and reduce inflammatory reactions. Based on current evidence about the pleiotropic effects of vitamin C, we hypothesize that the antioxidant properties of vitamin C might provide organ-protection and improve the kidney graft function in this model of ischemia and reperfusion.


METHODS
10 porcine kidneys from 5 Landrace pigs were perfused in vitro for 6 h. For each experiment, both kidneys of one animal were perfused simultaneously with a 1:1 mixture of autologous blood and modified Ringer's solution at 38 °C and 75 mmHg continuous perfusion pressure. One kidney was treated with a 500 mg bolus injection of vitamin C into the perfusate, followed by continuous infusion of 60 mg/h vitamin C. In the control test circuit, an equal volume of Ringer's solution was administered as a placebo. Perfusate samples were withdrawn at distinct points in time during 6 h of perfusion for blood gas analyses as well as measurement of serum chemistry, oxidative stress and antioxidant capacity. Hemodynamic parameters and urine excretion were monitored continuously. Histological samples were analyzed to detect tubular- and glomerular-injury.


RESULTS
vitamin C administration to the perfusate significantly reduced oxidative stress (49.8 ± 16.2 vs. 118.6 ± 23.1 mV; p = 0.002) after 6 h perfusion, and increased the antioxidant capacity, leading to red blood cell protection and increased hemoglobin concentrations (5.1 ± 0.2 vs. 3.9 ± 0.6 g/dL; p = 0.02) in contrast to placebo treatment. Kidney function was not different between the groups (creatinine clearance vit C: 2.5 ± 2.1 vs. placebo: 0.5 ± 0.2 mL/min/100 g; p = 0.9). Hypernatremia (187.8 ± 4.7 vs. 176.4 ± 5.7 mmol/L; p = 0.03), and a lower, but not significant decreased fractional sodium excretion (7.9 ± 2 vs. 27.7 ± 15.3%; p = 0.2) were observed in the vitamin C group. Histological analysis did not show differences in tubular- and glomerular injury between the groups.


CONCLUSION
Vitamin C treatment increased the antioxidant capacity of in vitro perfused kidney grafts, reduced oxidative stress, preserved red blood cells as oxygen carrier in the perfusate, but did not improve clinically relevant parameters like kidney function or attenuate kidney damage. Nevertheless, due to its antioxidative properties vitamin C might be a beneficial supplement to clinical kidney graft perfusion protocols.
['Bleilevens|Christian|C|', 'Doorschodt|Benedict M|BM|', 'Fechter|Tamara|T|', 'Grzanna|Tim|T|', 'Theißen|Alexander|A|', 'Liehn|Elisa A|EA|0000-0002-1253-3272', 'Breuer|Thomas|T|', 'Tolba|René H|RH|', 'Rossaint|Rolf|R|', 'Stoppe|Christian|C|', 'Boor|Peter|P|0000-0001-9921-4284', 'Hill|Aileen|A|0000-0002-7013-3357', 'Fabry|Gregor|G|']
D000818:Animals D000975:Antioxidants D001205:Ascorbic Acid D016207:Cytokines D005260:Female D006454:Hemoglobins D066298:In Vitro Techniques D007668:Kidney D009926:Organ Preservation D018384:Oxidative Stress D010477:Perfusion D015427:Reperfusion Injury D034421:Sus scrofa
vitamins antioxidant reperfusion injury organ dysfunction acute kidney injury oxidative stress animal models ascorbic acid kidney transplantation porcine kidney perfusion model in vitro techniques primary graft dysfunction