Figure 2.

Chloroquine inhibits NET formation in acute pancreatitis. Neutrophils harvested from mice with pancreatitis had evidence of spontaneous NET formation and greater propensity to form NETs upon stimulation with platelet activating factor (PAF), as visualized by staining of DNA with Hoechst (A). CQ treatment dramatically reduced spontaneous and stimulated NET formation. Supernatant DNA was measured to objectively quantify NETs (B). Serum cell-free DNA (C) and citrullinated histone H3 (CitH3) (D), biomarkers of in vivo NET formation were elevated with induction of pancreatitis, but significantly reduced with chloroquine treatment. Co-localization of pancreatic CitH3 (Green), with neutrophils (Ly-6G, Red) is increased in pancreatitis mice compared to sham controls (E), but treatment with CQ reduced neutrophil CitH3 expression (20x, scale bar = 50 μm). These are representative images from at least two independent analyses. ^*p < 0.05.