Acinous cell AR42J-derived exosome miR125b-5p promotes acute pancreatitis exacerbation by inhibiting M2 macrophage polarization via PI3K/AKT signaling pathway

Figure 8 miR-125b-5p promoted M1-type polarization of macrophages and the release of inflammatory factors. A: Construction of overexpression miR-125b-5p in RAW264.7 cell line; B: Negative control group: RAW264.7 cells stimulated with interleukin (IL)-4 for 24 h showed M2 polarization of macrophages; C: miR-125b-5p overexpression group: exosomes overexpressing miR-125b-5p were added to RAW264.7 cell line stimulated with IL-4 to inhibit M2 polarization of macrophages; D: miR-125b-5p overexpression group: Exosomes overexpressing miR-125b-5p were added to RAW264.7 cell line stimulated with IL-4 to promote M1 polarization of macrophages. Immunofluorescence counting showed that the number of M2 macrophages in the overexpression group was decreased compared with the control group; E: Western blot showed that the protein expression level of inducible nitric oxide synthase and CD206 in the RAW264.7 cell line; F: IL-6 and C-reactive protein level were determined by enzyme linked immunosorbent assay; G: miR-125b-5p overexpression promoted reactive oxygen species accumulation in the RAW264.7 cell line (P = 0.0224), ^aP < 0.05, ^bP < 0.005, ^cP < 0.0005. NC: Negative control; DAPI: 4',6-diamidino-2-phenylindole; iNOS: Inducible nitric oxide synthase; IL: Interleukin; CRP: C-reactive protein; ROS: Reactive oxygen species.