Therapeutic potential of exosome derived from hepatocyte growth factor-overexpressing adipose mesenchymal stem cells in TGFβ1-stimulated hepatic stellate cells

Abstract

Cirrhosis is a familiar end-stage of multiple chronic liver diseases. The gene-modified mesenchymal stem cells (MSCs) have become one of the most promising schemes for the treatment of cirrhosis. MSCs exhibit their therapeutic role mainly by secreting hepatocyte growth factor (HGF). The aim of this research was to probe the anti-fibrosis role of exosomes secreted by HGF modified-mouse adipose MSCs (ADMSCs) on activated hepatic stellate cells (HSCs) and to preliminarily explore the possible mechanism. Firstly, mouse ADMSCs were isolated and identified. Quantitative real-time polymerase chain reaction verified the transfection efficiency of ADMSC transfected with HGF lentivirus. Exosomes derived from ADMSC transfecting negative control/HGF (ADMSC^NC-Exo/ADMSC^HGF-Exo) were extracted by density gradient centrifugation. HSCs were allocated to the control, TGF-β, TGF-β + ADMSC-Exo, TGF-β + ADMSC^NC-Exo, and TGF-β + ADMSC^HGF-Exo groups. Moreover, all mice were distributed to the control, CCl₄ (40% CCl₄ in olive oil), CCl₄+ADMSC-Exo, CCl₄+ADMSC^NC-Exo, and CCl₄+ADMSC^HGF-Exo groups. Exosomes derived from ADMSCs with or without HGF transfection suppressed HSC activation, as evidenced by attenuating cell viability and cell cycle arrest at S phase but inducing apoptosis. Moreover, ADMSC-Exo, ADMSC^NC-Exo, and ADMSC^HGF-Exo effectively repressed the gene and protein levels of α-SMA, Col-I, Rho A, Cdc42, and Rac1 in TGF-β-treated HSCs, and ADMSC^HGF-Exo had the best effect. ADMSC^HGF-Exo had a stronger regulatory effect on serum liver index than ADMSC^NC-Exo in CCl₄-induced mice. In conclusion, ADMSC^HGF-Exo alleviated liver fibrosis by weakening the Rho pathway, thus reducing collagen production.