Immunofluorescence method to detect apoptotic events in the duodenum after a situation of hypoperfusion – preliminary results

Background: Apoptosis plays a key role in intestinal injury, as response to ischemia. In this study, apoptosis in the duodenum after haemorrhage and volume replacement was evaluated with M30, in a pig model. Material and methods: Apoptosis was evaluated in paraffin sections of duodenum from pigs that underwent total intravenous anaesthesia (TIVA) with propofol and remifentanil, and controlled bleeding and volume replacement using Ringer’s lactate (Group1, n=6). Apoptosis was also evaluated in a control group of animals that underwent TIVA, without bleeding (n=6). M30 Cytodeath immunofluorescence assay used a mouse monoclonal antibody (M30clone) (Roche) for detection of a caspase cleavage product of cytokeratin 18. Results: Group1: epithelial cells from the tips of the villi showed strong positive reaction, decreasing towards the body of the villi, negative in crypts and glands. Control group: epitelial cells from the tips of the villi showed a weak positive reaction, also decreasing towards the body and negative in crypts and glands. Discussion and Conclusions: Apoptosis occurred mainly in the epithelium of the villi, appearing to precede intestinal mucosa desquamation. Subsequent studies are being developed to determine the apoptotic index and correlation between immunohistochemical methods in small intestinal segments in the two groups of this study.FEDER-COMPETE Program, FCT, project COMPETE: FCOMP-01-0124-FEDER-009525. CI&DETS (PEst-OE/CED/UI4016/2014