Activation of PI3-kinase/Akt induced small bowel cell apoptosis during laparoscopic ischaemia-reperfusion of swine jejunum.
BACKGROUND
This study aimed to detect small bowel cell apoptosis by evaluating the nuclear activation of transcription factors, nuclear factor-kappaB (NF-kappaB) and Akt pathways as well as detecting apoptotic cells by TUNEL assay, during ischaemia-reperfusion (I/R) injury by Arterio-Venous Occlusion (AVO) of ajejunal loop in swine, using a laparoscopic procedure.


METHODS
A jejunal loop was isolated and a biopsy sample (T0) obtained. Biopsies were obtained at 30 minutes (T30I) and 60 minutes (T60I) after clamping and again during reperfusion (R), T30R and T60R. Standard H&E stains and immunohistochemical study for infiltrating polymorphonuclear leukocytes and TUNEL assays were performed. The activation of Akt and NF-kappaB in the animal model was evaluated by indirect immunofluorescence and confocal microscopy.


CONCLUSIONS
Our results confirmed that I/R injury of the jejunum did not only cause epithelial damage, but also activated the molecular pathways triggering an apoptotic reaction (by pro-inflammatory cytokines) and an anti-apoptotic mechanism that can lead to the regeneration of injured tissue.
Puglisi
F
F
General Surgery Unit, Department of Emergency and Organ Transplantation, Bari, Italy.
Lacitigniola
L
L
Loverre
A
A
Capuano
P
P
Martines
G
G
Staffieri
F
F
Rossi
G
G
Crovace
A
A
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D004195:Disease Models, Animal; D020287:In Situ Nick-End Labeling; D007583:Jejunum / Q000201:enzymology / Q000473:pathology* / Q000601:surgery; D010535:Laparoscopy; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015427:Reperfusion Injury / Q000201:enzymology* / Q000209:etiology / Q000473:pathology*; D013552:Swine; D000097782:NF-kappaB-Inducing Kinase
D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D000097782:NF-kappaB-Inducing Kinase