Specificity protein 1/ZIP8/zinc axis reduced macrophage extracellular traps to alleviate severe acute pancreatitis in mice

Figure 3 Macrophage extracellular trap formation depends on the peptidylarginine deiminase 4 pathway and reactive oxygen species, and M1 macrophages exhibit a strong ability to form macrophage extracellular traps. A and B: Primary macrophages were cocultured with phorbol 12-myristate 13-acetate and pancreatic acinar cells (following injury induced by caerulein), and the expression levels of double-stranded DNA (dsDNA) were subsequently measured in each group (n = 3); C and D: Representative immunofluorescence images of citrullinated histone H3 (CitH3) and F4/80 in macrophages from each group (C), as well as statistical analysis of the positivity rate of CitH3 (D), a macrophage extracellular trap marker (n = 5); E-H: After the administration of the peptidylarginine deiminase 4 inhibitor and anti-Ly6G, either alone or in combination, in the severe acute pancreatitis model, the levels of dsDNA in mouse plasma were measured (E), hematoxylin-eosin staining of pancreatic and lung tissues was performed (F), and the activities of amylase and lipase in plasma were determined (G and H) (n = 7); I and J: Representative immunofluorescence images of CitH3 and F4/80 in macrophages from each group, along with statistical analysis of CitH3 (n = 4). ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. NS: Not significant; MET: Macrophage extracellular trap; PMA: Phorbol 12-myristate 13-acetate; BMDMs: Bone marrow-derived macrophages; Nc: Negative control; dsDNA: Double-stranded DNA; CitH3: Citrullinated histone H3; Cl-amidine: BB-Cl-amidine; Ctrl: Control; DAPI: 4’,6-diamidino-2-phenylindole; NAC: N-Acetylcysteine; MFI: Mean fluorescence intensity.