Anti-inflammatory Properties of Ginseng-Derived Exosome-like Nanoparticles in LPS-induced RAW264.7

Abstract

Abstract Objectives Exosomes are extracellular nano-sized vesicles which transport proteins, lipids, nucleic acids and polysaccharides. Panax ginseng C.A. Meyer is known to have various pharmacological activities including anti-inflammatory and antioxidant effects. Furthermore, ginseng derived exosome-like nanoparticles (GDEs) may be considered to contribute to inflammation mechanisms. The main aim of this study elucidated that GDEs exert anti-inflammatory activities, which can molecularly modulate an inflammation mechanism in mammalian cells.Methods GDEs were isolated by the method using sucrose cushioning ultracentrifugation and the morphology of GDEs was shown through transmission electron microscope (TEM). The number of particles and size distribution of the GDEs were evaluated using nanoparticle tracking analysis (NTA). To investigate the anti-inflammatory activity of GDEs, the expression levels of inflammatory mediators and cytokines were analyzed in lipopoly-saccarides (LPS)-induced RAW 264.7, murine macrophages. ELISA, qPCR, WB and nitric oxide assay were carried out for analyzing anti-inflammatory effects of the GDEs in mammalian cells.Results GDEs showed a spherical shape with an average diameter of 176.2 nm and a substantial number of GDEs was counted as 1.09 x 1012 ± 1.19 x 1011 particles/ml. The pretreated GDEs in LPS-induced RAW264.7 suppressed the protein expression levels of pro-inflammatory cytokines including interlukin-1beta (IL-1β), interlukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α). In addition, protein expression levels of inflammation-related enzymes, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) as well as production of nitric oxide (NO) were dramatically decreased by the GDEs in LPS-induced RAW264.7. Correspondingly, the GDEs treatment clearly reduced LPS-induced mRNA expression levels of the iNOS and COX-2. In addition, expression patterns of the IL-1β, IL-6, TNF-α were gradually down-regulated by the GDEs in a dosage dependent manner.Conclusion These results demonstrated that Ginseng-derived exosome-like nanoparticles inhibited the expression of the inflammation mediators, suggesting that the GDEs possess anti-inflammatory activities and molecularly regulate the inflammatory mechanism in mammalian cells. Collectively, the GDEs has high potential as a novel therapeutic agent to treat chronic inflammatory diseases.