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30 April 2018 : Laboratory Research  

Cellular Mechanisms of Hepatoprotection Mediated by M2-Like Macrophages

Li Bai1ABCDE, Liming Fu2AD, Lu Li3BCF, Feng Ren4ADG, Qingfen Zheng1BCF, Shuang Liu1DE, Yuanping Han5ADG, Sujun Zheng1DG, Yu Chen1AG, Zhongping Duan1AG*

DOI: 10.12659/MSM.907222

Med Sci Monit 2018; 24: LBR2675-2682

Abstract

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 AND METHODS: Macrophages were isolated from control mice (M0 macrophages), then polarized into M1 or M2 phenotype using IFN-γ or IL-4, respectively. Conditioned media (CM) from M0, M1, 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 M0, M1, 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 M0 or M1 CM. Similarly, exposure of HL-7702 and HepG2 cells to either M0 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.

Keywords: Defense Mechanisms, Macrophage Activation

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750