An Experiment Research Of Autophagy Of Neural Stem Cell Induced By Oxidative Stress

[Objective]To research the induction of oxidative stress on autophagy of neural stemcells.[Methods] Separate and culture Wistar pregnant mice’s neural stem cells, and makethe good grow neural stem cell ball undergo Nestin immune cell chemical dyeing.Observe the differentiation of neural stem cells through immune cells chemicaltechnology. Detect the effect of H₂O₂on neural stem cells’ proliferation through MTTmethod. Observe the ultrastructure of autophagosome through electron microscopic.Make the NSCs in good condition undergo acridine orange （AO） dyeing and observethe outcome through fluorescence microscope. Detect the existence of LC3proteinthrough the Western blot technical.[Results] NSCs were successfully separated and cultured. The result by MTTcolorimetric shows that, under the effect of different doses of H₂O₂after12h or24h,the cell survival rates are gradually reduced with the increase in concentration of H₂O₂,and it shows dose-time correlation also. The comparison between0.5mM,1mMH₂O₂group and the control group has statistical significance. Compared with thecontrol group, red fluorescence positive results of0.5mM H₂O₂group aresignificantly enhanced, generates of cell acidic autophagy vacuoles are significantlyincreased. The autophagy vacuoles formation was observed by transmission electronmicroscopy after0.5mM H₂O₂acts on NSCs after12h. The result shows that, theNSCs performance of the control group displays the normal cell morphology, andnuclei are rules circular. The cell volume becomes large and expands under the effectof0.5mM H₂O₂, and autophagy vacuoles generate obviously. The expression of LC3protein of NSCs is detected by Western Blot under the effect of0.5mM H₂O₂roleafter12h, showing that LC3protein expression is significantly enhanced comparedwith the control group.[Conclusion]Oxidative stress can induce the occurrence of neural stem cells’autophagy.