Cellular Mechanisms of Hepatoprotection Mediated by M2-Like Macrophages

Background: Acute liver injury in the setting of hepatic fibrosis is an intriguing and still unsettled issue. We previously have demonstrated the protective effects conferred by M2-like macrophages in the fibrotic liver. In the present work, we further decipher the cellular mechanisms governing this hepatoprotection. Material/Methods: Macrophages were isolated from control mice (MO macrophages), then polarized into M1 or M2 phenotype us- ing IFN-gamma or IL-4, respectively. Conditioned media (CM) from MO, Ml, and M2 macrophages were harvested and applied to M1 macrophages. Cell apoptosis was evaluated by immunostaining and real-time PCR. Similarly, human monocyte-derived macrophages were isolated and polarized, then MO, Ml, and M2 CM were applied to HL-7702 or HepG2 cells followed by apoptosis induction. Cell apoptosis was assessed by flow cytometry. Results: For the mouse conditioned medium experiment, stronger expression of cleaved caspase 3 and higher Bax/Bcl- 2 mRNA ratio were found in M1 macrophages pretreated with M2 CM compared to those in M1 macrophages pretreated with MO or M1 CM. Similarly, exposure of HL-7702 and HepG2 cells to either MO or M1 CM had no significant effect on cell apoptosis. Nevertheless, the frequency of hepatocyte apoptosis was substantially reduced in HL-7702 (from 32.23 +/- 2.99 to 15.37 +/- 0.69 for Annexin V+/PI+ staining, p<0.01) and HepG2 cells (from 36.1 +/- 7.26 to 15.2+1.2 for Annexin V+/PI+ staining, p<0.01) with M2 CM pretreatment. Conclusions: M2-like macrophages exert their hepatoprotective effect by promoting M1-like macrophage apoptosis but protecting against hepatocyte apoptosis.