Association between High mobility group box-1 protein expression and cell death in acute pancreatitis

Abstract

The present study used caerulein stimulation of AR42J rat pancreatic cells as an in vitro acute pancreatitis (AP) model to investigate proteins differentially expressed in apoptosis and necrosis. AR42J cells were stimulated with 10-8mol/l caerulein and incubated for 24 h. Apoptosis and necrosis were detected using flow cytometry. The sorted Annexin V-positive cells (apoptotic) and the Annexin V/p ropidium iodide double-positive cells (necrotic) were analysed using proteomics. Results showed that numerous proteins were differentially expressed in these 2 groups. Functional enrichment analysis was performed on the differentially expressed genes using the Database for Annotation, Visualization and Integrated Discovery. High mobility group box-1 protein (HMGB1) wa s specifically expressed in the necrosis group. Models of varying degrees of AP were established using caerulein at concentrations of 10-9, 10-8, 10-7, 10-6 and 10-5 mol/l. The percentage of apoptotic and necrotic cells in each group was determined using flow cytometry. Protein expression levels of HMGB1 were detected by western blot analysis. The present study showed that as the concentration of caerulein increased, the percentage of necrotic cells and the protein expression levels of HMGB1 increased. HMGB1 is involved in many biological processes, including the chromosomal protein glycyl lysine isopeptide cross-link. HMGB1 may be involved in the early stage of pancreatitis, potentially by inducing the development of cell death by necrosis. These results provide an experimental basis for clinical intervention in A P.