Hypoxia-conditioned extracellular vesicles from human amniotic membrane stem cells attenuate D-galactose-induced aging in mice

초록

BACKGROUND Anti-aging for the preservation and recovery of physical and brain functions may be a major topic of regenerative medicine in the super-aging society. Stem cells and their secretory active molecules can play a central role of regenerative medicine. AIM To investigate whether extracellular vesicles (EVs) from amniotic membrane stem cells (AMSCs) enhance physical activity, including stamina, and cognitive function in a mouse model of facilitated brain-aging, and to elucidate underlying mechanisms. METHODS EVs were collected from conditioned media of AMSCs after hypoxic (2% O-2, 5% CO2) cultivation for 3 days. The size and composition of EVs was analyzed via nanoparticle-tracking analysis and proteome/lipidome profiles, and functional molecules such as growth factors and neurotrophic factors were analyzed via enzyme linked immunosorbent assay. Male ICR mice were subcutaneously administered with D-galactose (300 mg/kg) for 6 weeks to induce facilitated aging, during which intravenously injected with EVs (6 & times; 10(8) exosome particles/body) at weeks 0, 2, 4, and 6. Physical activity and cognitive function were assessed through Rota-rod, forced swimming and passive avoidance performances, respectively. To clarify underlying mechanisms, acetylcholine (ACh), brain-derived neurotrophic factor (BDNF), sirtuin 1 (SIRT1), glial fibrillary acid protein (GFAP), glycogen, and thiobarbituric acid-reactive substances (TBARS) were analyzed in the brain and muscles. RESULTS Six-week injection of D-galactose decreased physical activity and impaired learning and memory function, along with the reduced ACh, BDNF, SIRT1, and glycogen in the brain and muscles, whereas brain GFAP and muscular TBARS increased. However, EV treatment recovered the D-galactose-induced neurobehavioral deficits not only by increasing BDNF and SIRT1 (regulating neuro-muscular adaptation and function) and enhancing brain ACh (governing memory acquisition), but also by restoring muscular glycogen (an energy source) and attenuating brain GFAP and muscular TBARS (inflammatory and oxidative injury markers). CONCLUSIONEVs from AMSCs restored cognitive function of mice with facilitated brain-aging by increasing ACh, BDNF, and SIRT1. EVs also enhanced stamina not only by attenuating tissue injury, but also by strengthening the muscles through the production of glycogen and BDNF.

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