The Expression Of FIP200 On Retinoic Acid-induced Autophagy,differentiation And Apoptosis Of Neural Stem Cells

Objectives:From small to large concentrations of ATRA training NSCs, to study FIP200 and their interaction proteins levels on Retinoic acid-induced autophagy,differentiation and apoptosis,to initially confirmed NSCs’ different biological processes is by chaging the FIP200 protein level,also studied the relationship between three biological process,so as to provide a basis for further reveal how is ATRA affect nervous system diseases.Methods:Fetal rat cortical were isolated from 14.5 days pregnant C57 BL / 6J mice to cell culture and passaged.By microscopic morphology and differentiation immunofluorescence is neural stem cells.P3 NSCs add Different concentrations(0,0.1,0.5,1,5,10,15,20μM) induced culture24 h,AO staining to observe autophagy bubble,Semiquantitative PCR and Western blotting detection of LC3-II, FIP200 and P53 levels,to study the effect of retinoic acid in NSCs’ autophagy.Adding small doses of retinoic acid(0,0.1,0.5,1,5μM) induced cultured for7 days,Immunofluorescence assay the differentiation rate of NSCs, after adding LY294002 detection 1μM retinoic acid induce Differentiation change situation,research the Relationships about differentiation and autophagy in ATRA induce NSCs.Western blot to detect the FIP200 interacting proteins Rb1’s level.Adding Large doses of ATRA(5,10,15,20,40μM) induced culture 24 h,Using flow cytometry detect apoptosis rate, also detect FIP200 level.Add autophagy inhibitor LY294002 to Compare rate of apoptosis change,research the relationships about apoptosis and autophagy in ATRA induce NSCs.Results and:(1)Through morphology observation under a microscope and immunofluorescence results confirm is NSCs.(2)Different doses of ATRA cuture NSCs,FIP200 level is First rise after falling,the same to autophagy.P53 is the opposite.(3)Lower concentrations ATRA can induce NSCs differentiation.This is positively related with FIP200,Inhibition of autophagy can not inhibit differentiated,differentiation is related with RB1.(4)Large doses of ATRA make apoptosis.This is negative correlation to FIP200,Inhibition of autophagy can promote apoptosis.Conclusions:The effect of FIP200 on retinoic acid induced neural stem cells is to promote differentiation, anti-apoptotic.