Donor pretreatment with FK506 reduces reperfusion injury and accelerates intestinal graft recovery in rats.
BACKGROUND
FK506 alleviates warm ischemia-reperfusion injury, but it remains unknown if such protection is manifest after cold storage and transplantation. We studied the early outcome after transplantation of intestines from donors pretreated with FK506 compared to grafts from controls treated with saline (154 mM NaCl).


METHODS
Sprague-Dawley rats received 0.3 mg/kg FK506 or saline intravenously 6 hours before graft retrieval. The small bowel was harvested, stored for 3 hours, and then transplanted heterotopically. Samples were taken after preservation and at 20 minutes, 6 hours, 12 hours, and 24 hours after reperfusion. Heat shock protein 72 (Hsp72) and iintercellular adhesion molecule (ICAM)-1 expression and nuclear factor kappaB (NF-kappaB) activation were assessed via Western blots and eelectrophoretic mobility shift assay (EMSA), respectively. Dissacharidase activity and enterocyte proliferation rate were also studied.


RESULTS
Preservation injury was similar between groups, but pretreated grafts had better morphology already 20 minutes after reperfusion. Control grafts always had thinner mucosa and more PMN infiltration. Hsp72 expression was greater in pretreated grafts. ICAM-1 was absent after harvesting, preservation, and immediately after reperfusion but increased in control grafts at the later time points. Control grafts showed a biphasic NF-kappaB activation pattern, whereas NF-kappaB activation was inhibited effectively in pretreated grafts. Dissacharidase activity decreased during the first 6 hours after reperfusion but recovered within 24 hours in pretreated grafts but not in control grafts. Earlier enterocyte proliferation was observed in pretreated grafts.


CONCLUSIONS
FK506 donor pretreatment reduced graft proinflammatory activation and neutrophil inflammation. Pretreated groups revealed a milder reperfusion injury and accelerated morphologic and functional recovery. The mechanisms involved appear to involve Hsp72 upregulation and NF-kappaB inhibition.
Oltean
Mihai
M
Department of Surgery, Sahlgrenska Academy at Göteborg University, Göteborg, Sweden. mihai.oltean@wlab.gu.se
Pullerits
Rille
R
Zhu
Changlian
C
Blomgren
Klas
K
Hallberg
Eva C
EC
Olausson
Michael
M
D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D004186:Disaccharidases / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D020895:Enterocytes / Q000187:drug effects; D006085:Graft Survival / Q000187:drug effects; D050884:HSP72 Heat-Shock Proteins / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007421:Intestine, Small / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D008297:Male; D020556:Neutrophil Infiltration; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000473:pathology / Q000517:prevention & control*; D016559:Tacrolimus / Q000627:therapeutic use*; D014019:Tissue Donors; D016042:Transplantation, Heterotopic
D050884:HSP72 Heat-Shock Proteins; D007166:Immunosuppressive Agents; D018799:Intercellular Adhesion Molecule-1; D004186:Disaccharidases; D016559:Tacrolimus