Zm 241385

The preparation of the fibrin glue and the selection of the A2A receptor agonist and antagonist doses were based on the methods reported by several articles [27 (link)–30 (link)]. Briefly, fibrinogen powder (Solarbio, China) was dissolved in physiological saline into a concentration with 10 mg/ml at 37°C. Thrombin powder (Solarbio, China) was dissolved in 40 mmol/l calcium chloride solution, and the concentration was 20 μg/ml. Then, 5 mg CGS 21680 (specific adenosine A2A receptor agonist; Selleck, USA) or ZM 241385 (specific adenosine A2A receptor antagonist; Selleck, USA) was dissolved in 75 μl DMSO and mixed with thrombin solution into a concentration with 16 mg/ml CGS 21680 or ZM 241385 thrombin solution. Afterwards, 125 μl CGS 21680, ZM 241385, or thrombin solution (with DMSO as the solvent accelerating agent) was mixed with 125 μl fibrinogen solution to form a fibrin gel drug delivery system after 1 min of mixing.

The mice were given the following treatment, as once daily i.p. injections of ZM241385 (S8105; Selleckchem, USA) at a dose of 3 mg/kg or vehicle for three days. The injected volumes did not exceed 0.2 mL per animal. The doses of the drugs were based on previous studies [35 (link), 36 (link)].

Cerulein was purchased from MedChem Express (HY-A0190, Shanghai). Lipopolysaccharide (LPS) was purchased from Sigma (L2630, Shanghai). ABT702 was purchased from Merck Millipore (116890, Shanghai). ZM241385 (an adenosine A_2A receptor antagonist) and GSK2606414 (PERK inhibitor) were purchased from Selleck (S8105 and S7307, respectively, Shanghai). The following primary antibodies were purchased: ß-actin antibody (Proteintech, 66009), ADK antibody (Abcam, ab227087), RIP1 antibody (CST, 3,493), phosphor-RIP1 (Ser161) antibody (Affinity, AF7377), RIP3 antibody (CST, 15828), phosphor-RIP3 antibody (Abcam, ab195117), mouse MLKL antibody (Proteintech, 66675), rat MLKL antibody (Abcam, ab243142), phosphor-MLKL antibody (Affinity, AF7420), NF-κB p65 antibody (CST, 8242), phosphor-NF-κB p65 antibody (CST, 3,033), GRP78 Antibody (Proteintech, 11587), CHOP antibody (Proteintech, 15204), PERK antibody (Proteintech, 24390), phosphor-PERK antibody (CST, 3,179), eIF2α antibody (CST, 5324), phosphor-eIF2α antibody (CST, 3,398), CD11b antibody (Abcam, ab133357), MOMA-2 antibody (Abcam, ab33451), and amylase antibody (Santa Cruz Biotechnology, sc-46657). Secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, United States).