fig6

Figure 6. PgNVs induce proteomic variations on plasma EVs from AP mice and prevent NF-κB activation. (A) Representative images from TEM of plasma EVs from different mouse groups; (B) Representative images of NTA of plasma EVs from the different groups; (C) Determinations of particles/plasma ratio and mode size by NTA; (D) Particle/protein ratio, calculated as the number of particles per microgram of protein; (E) NF-κB levels in RAW 264.7 cells incubated with LPS (positive control), and EVs isolated from plasma of control, AP, and AP + PgNVs mice groups at 3 days post pancreatitis; (F) Volcano plots from proteomic analysis comparing AP EVs vs. control EVs; AP + PgNVs EVs vs. control EVs; and AP EVs vs. AP + PgNVs EVs. Red and blue dots represent proteins with P < 0.05 and significant Log2 fold change, respectively; while gray dots indicate non-significant differences; (G) Venn diagram displaying common and condition-specific proteins with statistically significant variations detected in plasma EVs isolated from AP, AP + PgNVs, and control mice groups; (H) GO classification of plasma EV proteins grouped as “biological process”, and LFQ intensity of proteins whose levels were significantly altered by AP or AP + PgNVs treatments. Measurements expressed as mean ± SD. *Significantly different from control group; ^#significantly different from AP, P < 0.05. AP: Acute pancreatitis; EVs: extracellular vesicles; PgNVs: pomegranate-derived nanovesicles; NF-κB: nuclear factor kappa B; TEM: transmission electron microscopy; NTA: nanoparticle tracking analysis; RAW: murine macrophage cell line RAW 264.7; LPS: lipopolysaccharide; LFQ: label-free quantification; GO: Gene Ontology; SD: standard deviation.