Mycoplasma gallisepticum induced exosomal gga-miR-193a to disturb cell proliferation, apoptosis, and cytokine production by targeting the KRAS/ERK signaling pathway

Int Immunopharmacol. 2022 Oct:111:109090. doi: 10.1016/j.intimp.2022.109090. Epub 2022 Jul 30.

Abstract

Mycoplasma gallisepticum (MG) is the main pathogen of chronic respiratory disease (CRD), an infectious disease in chickens with high morbidity. Exosomal miRNAs are emerging as important regulators in host immune response to microbial invasion. Previously, we found that gga-miR-193a was significantly up-regulated in exosomes from MG-infected primary chicken type II pneumocytes (CP-IIs). Therefore, the purpose of this study was to investigate the role of exosomal gga-miR-193a in MG infection. Exosomes were isolated and identified via ultracentrifugation, transmission electron microscopy, and nanoparticle-tracking analysis. Real-time quantitative PCR and Western blot were used to detect the gene expression. Enzyme-linked immunosorbent assay was used to examine the levels of the inflammatory cytokines. CCK-8 and flow cytometry assays were applied to analyze the cell functions. The results showed that MG infection induced high expression of gga-miR-193a in exosomes from CP-IIs. Moreover, exosomes secreted by MG-infected CP-IIs could selectively transport gga-miR-193a into DF-1 cells. Exosomal gga-miR-193a internalized by DF-1 cells inhibited cell proliferation, promoted apoptosis, and increased interleukin-1β and tumor necrosis factor-α secretions by targeting the RAS/ERK signaling pathway. These results suggest that MG induced the secretion of gga-miR-193a by exosomes to damage the life activities of normal cells, which partially interpreted the mechanism of MG establishing systemic chronic infection in the body.

Keywords: Cell life activity; Exosomes; Mycoplasma gallisepticum; RAS/ERK signaling pathway; gga-miR-193a.

MeSH terms

  • Animals
  • Apoptosis
  • Cell Line
  • Cell Proliferation
  • Chickens
  • Cytokines / metabolism
  • Fibroblasts / metabolism
  • Gene Expression
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Mycoplasma Infections* / genetics
  • Mycoplasma Infections* / metabolism
  • Mycoplasma Infections* / veterinary
  • Mycoplasma gallisepticum* / genetics
  • Mycoplasma gallisepticum* / metabolism
  • Proto-Oncogene Proteins p21(ras) / metabolism
  • Signal Transduction

Substances

  • Cytokines
  • MicroRNAs
  • Proto-Oncogene Proteins p21(ras)