Leucocyte depletion improves renal function in porcine kidney hemoreperfusion through reduction of myeloperoxidase+ cells, caspase-3, IL-1β, and tubular apoptosis.
BACKGROUND
Leucocytes play crucial roles in ischemia reperfusion injury that is inevitable in kidney transplantation. Leucocyte-depleted hemoreperfusion improved post-ischemic renal function was demonstrated in our previous study and its underlying mechanisms were further investigated in this study.


METHODS
Porcine kidneys were subjected to 7 min warm ischemia and 2 h cold storage, and preserved by hemoreperfusion with or without leucocyte depletion for 6 h on an isolated organ perfusion system.


RESULTS
Tubulointerstitial damage was improved by leucocyte depletion, which was accompanied by reduced myeloperoxidase+ cell infiltration up to 91%. Apoptotic cells in tubular and interstitial areas were increased by hemoreperfusion, but tubular apoptosis was decreased by leucocyte depletion. The raised caspase-3 activity by hemoreperfusion was almost completely abolished by leucocyte depletion. In addition, the expression of IL-1β active subunit was enhanced by hemoreperfusion, but partially reduced by leucocyte depletion, although IL-1β precursor and HSP70 were increased by hemoreperfusion regardless of leucocyte depletion. Furthermore, myeloperoxidase+ cells were associated with caspase-3 activity, both of which were positively correlated with tubular apoptosis, IL-1β active subunit, tubulointerstitial damage, and serum creatinine, while HSP70 was linked to renal blood flow.


CONCLUSIONS
Leucocyte depletion improved post-ischemic renal function and structure was mainly due to reduced infiltration of myeloperoxidase+ cells, which was associated with decreased apoptosis, caspase-3 activity and IL-1β activation.
Yang
Bin
B
Transplant Group, Department of Infection, Immunity and Inflammation, University of Leicester, Leicester General Hospital, University Hospitals of Leicester, Leicester, United Kingdom. by5@le.ac.uk
Hosgood
Sarah A
SA
Harper
Simon J F
SJ
Nicholson
Michael L
ML
D000818:Animals; D017209:Apoptosis; D002315:Cardiopulmonary Bypass; D053148:Caspase 3 / Q000378:metabolism; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006464:Hemoperfusion; D053583:Interleukin-1beta / Q000378:metabolism; D007511:Ischemia; D007668:Kidney / Q000201:enzymology / Q000502:physiology*; D007677:Kidney Function Tests; D007684:Kidney Tubules / Q000502:physiology; D007958:Leukocyte Count; D047589:Leukocyte Reduction Procedures / Q000379:methods; D009926:Organ Preservation / Q000379:methods; D009195:Peroxidase / Q000378:metabolism; D015427:Reperfusion Injury / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control; D013552:Swine
D018840:HSP70 Heat-Shock Proteins; D053583:Interleukin-1beta; D009195:Peroxidase; D004253:DNA Nucleotidylexotransferase; D053148:Caspase 3