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CB2R agonist GW405833 alleviates acute liver failure in mice via inhibiting HIF-1α-mediated reprogramming of glycometabolism and macrophage proliferation

Abstract

The inflammatory responses involving infiltration and activation of liver macrophages play a vital role in acute liver failure (ALF). In the liver of ALF mice, cannabinoid receptor 2 (CB2R) is significantly upregulated on macrophages, while CB2R agonist GW405833 (GW) could protect against cell death in acute liver damage. In this study, we investigated the molecular mechanisms underlying the protective effects of GW against ALF in vivo and in vitro from a perspective of macrophage glycometabolism. Mice were pretreated with GW (10 mg/kg, i.p.), then were injected with D-GalN (750 mg/kg, i.p.) and LPS (10 mg/kg, i.p.) to induce ALF. We verified the protective effects of GW pretreatment in ALF mice. Furthermore, GW pretreatment significantly reduced liver macrophage infiltration and M1 polarization, and inhibited the release of inflammatory factors TNF-α and IL-1β in ALF mice. These protective effects were eliminated by CB2R antagonist SR144528 or in CB2R−/− ALF mice. We used LPS-stimulated RAW264.7 cells as an in vitro M1 macrophage-centered model of inflammatory response, and demonstrated that pretreatment with GW (10 μM) significantly reduced glucose metabolism by inhibiting glycolysis, which inhibited LPS-induced macrophage proliferation and inflammatory cytokines release. We verified these results in a stable CB2R−/− RAW264.7 cell line. Moreover, we found that GW significantly inhibited the expression of hypoxia inducible factor 1α (HIF-1α). Using a stable HIF-1α−/− RAW264.7 cell line, we confirmed that GW reduced the release of inflammatory cytokines from macrophages and inhibited glycolysis by downregulating HIF-1α expression. In conclusion, activation of CB2Rs inhibits the proliferation of hepatic macrophages and release of inflammatory factors in ALF mice through downregulating HIF-1α to inhibit glycolysis.

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Fig. 1: Survival analysis of grouped mice.
Fig. 2: Protective effect of GW in ALF.
Fig. 3: Effects of CB2R activation on macrophage activation, inflammation, and apoptosis of liver cells in ALF.
Fig. 4: Effect of CB2R activation on macrophage proliferation and release of inflammatory cytokines.
Fig. 5: Effect of CB2R activation on glucose metabolism in LPS-activated macrophages.
Fig. 6: Effects of CB2R knockdown for LPS-activated macrophages on cell proliferation, release of inflammatory factors and glucose metabolism.
Fig. 7: Effect of HIF-1α knockdown on GW activated CB2R to alter LPS-stimulated macrophages.
Fig. 8: Differential liver damage in WT and CB2R−/− mice.
Fig. 9: Differential expression of HIF-1α in liver macrophages of ALF in WT and CB2R−/− mice.

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (81700561, 81970523), National Science and Technology Major Project (2018ZX10723203), National Natural Sciences Foundation of Hunan province (2019JJ40496, 2020JJ4877, 2020JJ4879). Sources of funding did not have any role in study design, data collection or analysis, interpretation of results, or manuscript preparation or submission. The corresponding author confirms that all the authors have full access to the full data in the study and accept responsibility to submit for publication.

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ZBH: Conceptualization, project design, and guidance, Funding acquisition, project administration. XGF, JW, YW: writing—review & editing. SLC: Writing—original draft, data curation, formal analysis, visualization. SYP, YXZ, JC: Methodology, investigation. XWH, YH, NL: Writing—review & editing, methodology. All authors read and approved the final version of the manuscript.

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Correspondence to Ze-bing Huang.

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Cai, Sl., Fan, Xg., Wu, J. et al. CB2R agonist GW405833 alleviates acute liver failure in mice via inhibiting HIF-1α-mediated reprogramming of glycometabolism and macrophage proliferation. Acta Pharmacol Sin (2023). https://doi.org/10.1038/s41401-022-01037-8

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  • DOI: https://doi.org/10.1038/s41401-022-01037-8

Keywords

  • acute liver failure
  • cannabinoid receptor 2
  • glycolysis
  • macrophages
  • RAW264.7 cells
  • hypoxia-inducible factor 1α
  • GW405833
  • SR144528

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