Ozone promotes macrophage efferocytosis and alleviates neuropathic pain by activating the AMPK/Gas6-MerTK/SOCS3 signaling pathway

Front Immunol. 2024 Nov 19:15:1455771. doi: 10.3389/fimmu.2024.1455771. eCollection 2024.

Abstract

Background: Neuropathic pain (NPP) is a multifaceted pain syndrome that occurs as a consequence of physical injury or underlying diseases, with an incidence rate of 7%-10%, NPP poses a significant clinical challenge as current treatment options are ineffective. The accumulation of apoptotic cells and neuroinflammation play crucial roles in the pathological mechanisms of NPP. Here, we aim to investigate strategies for effectively clearing apoptotic cells and provide therapeutic interventions for NPP.

Methods: CCI mice were treated with different concentrations of ozone (15μg, 30μg, 45μg) to investigate the effects on the accumulation of apoptotic cells and neuroinflammation. In vitro, the phagocytic function of BMDM towards apoptotic neutrophils after ozone treatment was examined.

Results: We found ozone at a concentration of 30μg significantly alleviated mechanical hypersensitivity in CCI mice and ozone significantly upregulates the phagocytic activity of BMDM. Furthermore, we investigated the mechanisms and found ozone could activate AMPK, upregulate Gas6 (but not Protein S), activate MerTK (a key receptor involved in apoptosis), and enhance the phagocytic function of BMDM towards apoptotic neutrophils. It caused the promotion of SOCS3 expression and the suppression of inflammatory factors IL-1β, IL-6, and TNF-a. Interestingly, the effect of ozone in alleviating CCI-induced pain was abolished by the AMPK inhibitor CC and the MerTK receptor inhibitor UNC2541.

Conclusion: Ozone facilitated macrophage clearance of apoptotic cells, decreased neuroinflammation by activation of p-AMPK/Gas6/MerTK/SOCS3 signaling pathway, which may become an effective therapeutic approach for neuropathic pain after further clinical validation.

Keywords: MerTK; SOCS3; efferocytosis; macrophage; neuroinflammation; neuropathic pain; ozone.

MeSH terms

  • AMP-Activated Protein Kinases / metabolism
  • Animals
  • Apoptosis / drug effects
  • Disease Models, Animal
  • Efferocytosis
  • Intercellular Signaling Peptides and Proteins* / metabolism
  • Macrophages / drug effects
  • Macrophages / immunology
  • Macrophages / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neuralgia* / drug therapy
  • Neuralgia* / etiology
  • Neuralgia* / metabolism
  • Ozone*
  • Phagocytosis* / drug effects
  • Signal Transduction* / drug effects
  • Suppressor of Cytokine Signaling 3 Protein* / metabolism
  • c-Mer Tyrosine Kinase* / metabolism

Substances

  • Suppressor of Cytokine Signaling 3 Protein
  • c-Mer Tyrosine Kinase
  • Ozone
  • growth arrest-specific protein 6
  • Mertk protein, mouse
  • Socs3 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • AMP-Activated Protein Kinases

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the National Natural Science Foundation of China (82271252, 82204542, and 81971047), the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No. 21KJB310019), Jiangsu Joint Project of Ili & Jiangsu Joint Institute of Health (yl2022ms02).