Changes of PACAP immunoreactivities and cytokine levels after PACAP-38 containing intestinal preservation and autotransplantation.
Small bowel is one of the most sensitive organs to ischemia-reperfusion injury, which is a significant problem during transplantation. Pituitary adenylate cyclase-activating polypeptide (PACAP) has cytoprotective effect in ischemic injuries of various tissues. The aim of our study was to measure changes of PACAP-38 and PACAP-27 immunoreactivities and cytokine levels in intestinal grafts stored in PACAP-38-containing preservation solution. Small bowel autotransplantation was performed on male Wistar rats. Grafts were stored in University of Wisconsin (UW) solution at 4 °C for 1 h (group (G)I), for 3 h (GII), and for 6 h (GIII) and in PACAP-38-containing UW solution for 1 h (GIV), for 3 h (GV), and for 6 h (GVI). After preservation, performing vessel anastomosis reperfusion began, which lasted 3 h in each group. Tissue biopsies were collected after laparotomy (control) and at the end of the reperfusion periods. Intestinal PACAP-38 and PACAP-27 immunoreactivities were measured by radioimmunoassay. To measure cytokines from tissue homogenates, we used rat cytokine array and Luminex Multiplex Immunoassay. Levels of PACAP-38 and PACAP-27 immunoreactivity decreased after 1 and 3 h preservation compared to control levels. This decrease was significant following 6 h cold storage (p < 0.05). Values remained significantly higher in grafts stored in PACAP-38-containing UW. Cytokine array revealed that expression of the soluble intercellular adhesion molecule-1 (CD54) and L-selectin (CD62L/LECAM-1) was increased in GIII. Both 6 h cold storage in PACAP-38-containing UW solution and 3 h reperfusion caused strong reduction in these cytokines activation in GVI. RANTES (CCL5) levels were increased in all groups. Strong activation of the tissue inhibitor of metalloproteinase-1 was in GIII. However, PACAP-38-containing cold storage could decrease its activation in GVI. Furthermore, strong activation of the tissue inhibitor of metalloproteinase-1 was detected in 6 h preserved grafts without PACAP-38 (GIII). PACAP-38-containing cold storage could decrease its activation in GVI. Our present study showed that PACAP-38 and PACAP-27 immunoreactivities decreased in a time-dependent manner during intestinal cold preservation, which could be ameliorated by administration of exogenous PACAP-38 to the preservation solution. Moreover, PACAP-38 could attenuate tissue cold ischemic injury-induced changes in cytokine expression.
Department of Surgical Research and Techniques, University of Pecs, Pecs, Hungary.
Klara
K
Nedvig
Gyorgy
G
Weber
Jozsef
J
Nemeth
Krisztina
K
Kovacs
Dora
D
Reglodi
Agnes
A
Kemeny
Andrea
A
Ferencz
D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D018946:Chemokine CCL5 / Q000032:analysis / Q000096:biosynthesis; D003080:Cold Temperature; D016207:Cytokines / Q000032:analysis* / Q000096:biosynthesis; D005978:Glutathione / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis / Q000096:biosynthesis; D007421:Intestine, Small / Q000737:chemistry / Q000378:metabolism / Q000637:transplantation*; D019041:L-Selectin / Q000032:analysis / Q000096:biosynthesis; D007813:Laparotomy; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000032:analysis* / Q000494:pharmacology; D020033:Protein Isoforms / Q000032:analysis; D011863:Radioimmunoassay; D011887:Raffinose / Q000494:pharmacology; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D015424:Reperfusion; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis / Q000096:biosynthesis; D014182:Transplantation, Autologous
D018946:Chemokine CCL5; D016207:Cytokines; D007328:Insulin; D019188:Organ Preservation Solutions; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D020033:Protein Isoforms; C576839:TIMP1 protein, rat; D019715:Tissue Inhibitor of Metalloproteinase-1; C055206:University of Wisconsin-lactobionate solution; D018799:Intercellular Adhesion Molecule-1; D019041:L-Selectin; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose