Study of cell death in duodenal mucosa in celiac disease

CAROLINA N. RUERA; FEDERICO PEREZ; LUCIANA GUZMAN; LORENA MENENDEZ; LAURA GARBI; FERNANDO CHIRDO

Ayr. Escocia

Encuentro; 32rd Meeting of the European Prolamin Working Group of Prolamin Analysis and Toxicity; 2018

European Prolamin Working Group of Prolamin Analysis and Toxicity

Celiac disease (CD) is a chronic enteropathy characterized by massive loss of enterocytes from proximal small intestine. Apoptosis has been considered as the main pathway of enterocytes death, however this mechanism has been only partially studied.The aim of this work was to characterize the cell death pathways in intestinal mucosa in CD enteropathy.Duodenal biopsies were collected from pediatric and adult patients during the routine procedure for CD diagnosis. Ethic committees from Public Health Institutions approved this work. Expression of caspase 1, 3, 8, 4 and Gasdermine D (GSDMD) were assessed by Western blot (WB) in protein extracts from whole biopsy. TUNEL reaction and Immunofluorescence microscopy (IFI) analysis were performed on sections of paraffin-embedded tissues.TUNEL reaction, which accounts for an advanced stage in cell death, showed increased number of TUNEL+ cells of duodenal lamina propria of CD patients compared with healthy controls.WB analysis of protein extracts from biopsies of CD patients showed higher expression of Caspase 3. In addition, IFI analysis showed significative increment in caspase 3 and 8 in epithelium and lamina propria of duodenal samples of CD patients.Moreover, expression of Caspase 1 and GSDMD determined by WB were also significantly increased in active CD patients. In conclusion, we found that multiple pathways associated with cell death and inflammation are activated in the small intestine of CD patients at diagnosis. These findings suggest that apoptosis, as well as pyroptosis, may occur jointly in the enteropathy.