Protective Effect Of Purify Exosomes From Human Adipose-derived Mesenchymal Stem Cells On Glutamate Excitotixicity-induces Damage In PC12 Cells

Objective: To observe whether the human adipose mesenchymal stem cells derived-exosomes had protective effects on excitotoxicity-induced damage of PC12 cell model and discuss its possible mechanism.Methods: The experiment was divided into two main parts,first we culture adipose derived mesenchymal stem cells(AMSCs)and collect the conditioned medium.Then extract exosomes by ultracentrifugation and identify the vesicles by transmission electron microscopy technology.Then glutamate excitotoxicity model of PC12 cells was established to evaluate the effects of AMSC-exosomes.To measure the death rate of PC12 cells,we use LDH release and trypan blue staining.In the second part we isolated and cultured neonatal SD rat cortical neurons,identified purity of cultured neurons by MAP-2.Establish glutamate excitotoxicity model of PC12 cells and neurons,add different concentrations of AMSC-exosomes and test LDH release.Then detect the energy metabolism of PC12 cells and its autophagy pathway in order to reveal the mechanism of the cell protection.Results: After ultracentrifugation,exosomes were found in the conditional medium from adipose derived mesenchymal stem cells,LDH release proved that AMSC-exosomes can effectively protect PC12 cells from glutamate excitotoxicity.NAD/NADH result showed that the protective effect has no significant relationship with energy metabolism,but exosomes can enhance cell autophagy to against glutamate injury in PC12 cells.Conclusions:Purify exosomes from human adipose-derived mesenchymal stem cells can effectively reduce LDH release rate of PC12 cell model and protect PC12 cells from glutamate excitotixicity-induces damage.Purify exosomes from human adipose-derived mesenchymal stem cells has no significant protective effect on glutamate excitotixicity-induces damaged neures.The protective effect of purify exosomes from human adipose-derived mesenchymal sem cells on glutamate excitotixicity-induces damage in PC12 cells has no significant relationship with energy metabolism,but associated with cell autophagy.