Activation of PI3-Kinase/Akt Induced Small Bowel Cell Apoptosis during Laparoscopic Ischaemia-Reperfusion of Swine Jejunum

Abstract

Introduction: This study aimed to detect small bowel cell apoptosis by evaluating the nuclear activation of transcription factors, nuclear factor-kB (NF-kB) and Akt pathways as well as detecting apoptotic cells by TUNEL assay, during ischaemia-reperfusion (I/R) injury by Arterio-Venous Occlusion (AVO) of a jejunal loop in swine, using a laparo-scopic procedure.

Materials and 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-kB 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.

Key Words:

• Cell apoptosis
• ischemia-reperfusion
• laparoscopy
• jejunum
• experimental surgery.

Additional information

Notes on contributors

G. Martines

G. Martines, M.D. Via Giuseppe Pavoncelli 139 70125 Bari, Italy Fax: [phone] E-mail :[email protected]