Mechanism of ADSC-Exos promoting macrophage migration via Itgav/Fak/Src signaling pathway

Abstract Macrophages play an important role in peripheral nerve regeneration, but the specific mechanisms remain unclear. Our previous studies demonstrated that adipose stem cell-derived exosomes (ADSC-Exos) promote peripheral nerve regeneration, correlating with their effects on macrophages. To test this, ADSC-Exos were isolated from primary ADSCs and co-cultured with RAW264.7 macrophages. We examined ADSC-Exos' impact on macrophage migration by assessing RAW264.7 cell viability, migration capability, and adhesion ability. Transcriptome sequencing identified differentially expressed genes (DEGs) and examined their interactions. Key gene expressions were verified by qRT-PCR, while pathway-related proteins were detected using Western blot and qRT-PCR. Results showed ADSC-Exos enhanced RAW264.7 cell viability, adhesion and migration. Transcriptomic analysis revealed 233 DEGs. GO analysis showed DEGs were enriched in biological processes including negative regulation of damage response and cell migration, cellular constituents like cytoskeleton and adhesion complexes, and molecular functions including redox activity and ubiquitination. KEGG analysis indicated DEGs were abundant in pathways of cell adhesion molecules, MAPK, folate-dependent one-carbon metabolism, and NF-κB. ADSC-Exos promoted Itgav expression and Fak, Src, and Bcar1 phosphorylation. The Itgav inhibitor cilengitide decreased Itgav expression and phosphorylation of Fak, Src, and Bcar1. These findings suggest ADSC-Exos enhance macrophage migration and adhesion through the Itgav/Fak/Src signaling pathway.