Molecular Therapy for Non-Alcoholic Fatty Liver Disease: Angiotensin-(1-7) Delivery via Cyclic RGD-Modified Vesicles Activates Mas Receptor to Ameliorate Fibrosis Through Autophagy and Metabolic Reprogramming

Non-alcoholic fatty liver disease (NAFLD) is a global health burden characterized by hepatic steatosis and progressive fibrosis, necessitating novel therapeutic strategies. This study investigates the molecular mechanism by which cyclic RGD peptide (cRGD)-modified adipose-derived mesenchymal stem cell (ADMSC)-derived extracellular vesicles (EVs) deliver Angiotensin-(1-7) to attenuate NAFLD-associated liver fibrosis. EVs were isolated from murine ADMSCs via ultracentrifugation, surface-modified with cRGD using EDC/NHS crosslinkers, and loaded with Angiotensin-(1-7) via an ultrasound-assisted method. The therapeutic effects were evaluated in vitro using hepatic stellate cells (LX-2) and in vivo using a high-fat diet (HFD)-induced NAFLD mouse model. Multi-omics analyses (transcriptomics, proteomics, metabolomics) were performed on liver tissues to elucidate underlying pathways. Results demonstrated that cRGD-modified EVs loaded with Angiotensin-(1-7) exhibited excellent biocompatibility and targeted liver accumulation, significantly reducing hepatic lipid accumulation, fibrosis, and serum markers of liver damage (ALT, AST). Mechanistically, Angiotensin-(1-7) activated the Mas receptor, enhancing Akt-Foxo1-dependent Autophagy and fatty acid metabolism reprogramming, as confirmed by upregulation of autophagy-related proteins (LC3-II, p62) and downregulation of fibrosis markers (TGF-β1, α-SMA, Collagen I). Multi-omics data revealed enrichment in fatty acid degradation and Autophagy pathways, while Mas receptor inhibition abolished these effects. This study establishes that cRGD-modified EVs deliver Angiotensin-(1-7) as a potent strategy to mitigate NAFLD fibrosis through Mas/Akt/FOXO1 signaling, offering a promising therapeutic avenue for metabolic liver diseases.

adipose‐derived mesenchymal stem cells; angiotensin‐(1–7); autophagy; cyclic RGD peptide; extracellular vesicles; non‐alcoholic fatty liver disease.