Huc-MSCs-derived exosomes attenuate neuropathic pain by inhibiting activation of the TLR2/MyD88/NF-κB signaling pathway in the spinal microglia by targeting Rsad2

Int Immunopharmacol. 2023 Jan:114:109505. doi: 10.1016/j.intimp.2022.109505. Epub 2022 Dec 11.

Abstract

Background: Mesenchymal stem cells (MSCs)-derived exosomes have shown promise as a cell-free therapeutic strategy for neuropathic pain. This study was conducted to explore the potential mechanisms underlying the analgesic effects of MSC-derived exosomes in treating neuropathic pain.

Methods: Human umbilical cord MSCs (huc-MSCs)-derived exosomes were isolated and identified. BV-2 microglia were stimulated with lipopolysaccharide (LPS) in the presence or absence of exosomes. Differentially expressed proteins were identified by tandem mass tag (TMT)-based proteomic analysis. The analgesic effects of huc-MSCs-derived exosomes were evaluated in a rat model of chronic constriction injury (CCI). The underlying mechanism was investigated by flow cytometry, RT-qPCR, Western blotting, immunofluorescent staining, and small interfering RNA transfection.

Results: In vitro, huc-MSCs-derived exosomes suppressed LPS-induced microglial activation and inhibited activation of the TLR2/MyD88/NF-κB signaling pathway. Based on the proteomic analysis, Rsad2 was identified and confirmed to be down-regulated by huc-MSCs-derived exosomes. Importantly, knockdown of Rsad2 also inhibited microglial activation and restrained activation of the TLR2/MyD88/NF-κB signaling pathway. In vivo, intrathecal injection of exosomes ameliorated CCI-induced mechanical allodynia, down-regulated Rsad2 expression and restrained TLR2/MyD88/NF-κB signaling activation in the spinal microglia.

Conclusion: Huc-MSCs-derived exosomes exerted analgesic effects on neuropathic pain by inhibiting activation of the TLR2/MyD88/NF-κB signaling pathway in the spinal microglia. The mechanism underlying these antinociceptive effects involved exosome-mediated interference with Rsad2 expression, thereby inhibiting microglial activation.

Keywords: Huc-MSCs-derived exosomes; Microglial activation; Neuropathic pain; Rsad2; TLR2.

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Analgesics / pharmacology
  • Animals
  • Exosomes* / metabolism
  • Humans
  • Lipopolysaccharides / pharmacology
  • Mesenchymal Stem Cells* / metabolism
  • Microglia / metabolism
  • Myeloid Differentiation Factor 88 / genetics
  • Myeloid Differentiation Factor 88 / metabolism
  • NF-kappa B / metabolism
  • Neuralgia* / drug therapy
  • Oxidoreductases Acting on CH-CH Group Donors* / metabolism
  • Oxidoreductases Acting on CH-CH Group Donors* / pharmacology
  • Oxidoreductases Acting on CH-CH Group Donors* / therapeutic use
  • Proteomics
  • Rats
  • Signal Transduction
  • Toll-Like Receptor 2 / genetics
  • Toll-Like Receptor 2 / metabolism

Substances

  • NF-kappa B
  • Toll-Like Receptor 2
  • Myeloid Differentiation Factor 88
  • Lipopolysaccharides
  • Adaptor Proteins, Signal Transducing
  • Analgesics
  • TLR2 protein, human
  • RSAD2 protein, human
  • Oxidoreductases Acting on CH-CH Group Donors
  • Myd88 protein, rat