Netrin-1 enriched ADSC exosomes attenuate abdominal aortic aneurysm via NF-κB and JNK/AP-1 modulation

Background Abdominal aortic aneurysm (AAA) is a macrophage-driven disease without effective pharmacological therapy. Given the anti-inflammatory effects of Netrin-1 in macrophages, this study aimed to investigate whether exosomes derived from Netrin-1 overexpressing adipose-derived stem cells (N-ADSC-exo) can reprogram macrophages and attenuate AAA. Methods Murine ADSCs were transduced with or without Netrin-1 adenoviral vectors to generate ADSC-exo and N-ADSC-exo. Effects on RAW264.7 macrophages were evaluated by adhesion and migration assays, LPS-induced ROS, apoptosis, and CCK-8 viability. Bulk RNA sequencing with GO/KEGG enrichment and GSEA compared transcriptomes. NF-κB and JNK/AP-1 signaling and polarization markers were analyzed by Western blotting. In ApoE −/− mice with Ang II-induced AAA, PBS, ADSC-exo, or N-ADSC-exo were injected intravenously. AAA incidence, maximal abdominal aortic diameter, elastin/collagen histology, immunofluorescence analysis of macrophage polarization, aortic IL-1β/MMP9, DiD-labeled exosome biodistribution, and systemic biocompatibility were assessed. Results ADSC-exo and N-ADSC-exo were similarly internalized by macrophages. Compared with ADSC-exo, N-ADSC-exo enhanced adhesion and migration, reduced LPS-induced ROS without increasing apoptosis, and modestly increased viability. RNA sequencing showed enrichment of cell-cycle pathways and suppression of NF-κB related gene sets in N-ADSC-exo treated cells, consistent at the protein level with decreased p-p65, p-JNK, AP-1 and IL-1β, and increased M2 markers. In AAA mice, N-ADSC-exo showed preferential uptake by F4/80 + cells, lowered AAA incidence, reduced aortic diameter, improved elastin preservation, decreased F4/80 + CD86 + macrophages and IL-1β/MMP9, and did not induce systemic toxicity. Conclusions N-ADSC-exo mitigate AAA via macrophage reprogramming toward a NF-κB and JNK/AP-1 suppressed, M2-skewed phenotype.