Induction and Treatment of Acute Pancreatitis Models

TLCS-AP was induced by pancreatic ductal infusion of 3 mM TLCS at 5 μL/min for 10 min via an infusion pump as described previously [20 (link)]; FAEE-AP was induced by 2 hourly intraperitoneal injections of 1.35 g/kg ethanol and 150 mg/kg palmitoleic acid (POA) [4 (link)], and mice sacrificed 24 h later. CER-AP was induced by 7 hourly intraperitoneal cerulein injections at a dose of 50 mg/kg [3 (link)] and mice sacrificed 12 h later. In all models, analgesia was administered using 0.1 mg/kg buprenorphine hydrochloride (Temgesic, Reckitt and Coleman, Hull, UK). All experimental protocols were approved by the local ethics committee (University of Liverpool). Nec-1 (Sigma-Aldrich, N9073, Dorset, UK) was dissolved in 10% DMSO + 90% PEG 400 and administered at a dose of 56 mg/kg consistently for 12/24 h via subcutaneous osmotic mini-pump (Charles River UK, Ltd., ALZET osmotic mini-pumps (2001D), Cambridge, UK). Epacadostat (EPA) (Selleck Chemicals, INCB24360, Cambridge, UK) was dissolved in 10% DMSO + 90% PEG 400 and delivered through osmotic mini pump at the dose of 50 mg/kg [21 (link)]. Osmotic mini-pumps for treatments (Nec-1, EPA) were inserted into the mice 0.5 h after TLCS-AP induction, after the 2nd injection of POA in FAEE-AP and after the 3rd injection of caerulein in CER-AP [22 (link)].