ABSTRACT
ABSTRACT:Objective To explore the role of PTEN in the suppression of neural stem cells so as to clarify whether neural stem cell proliferation can be promoted by regulating the PI3K-Akt/PTEN expression level. Methods We removed the hippocampus from neonatal 24 h Kunming mice,isolated and cultured the generation of neural stem cells,which were then identified by immunofluorescence test.We randomly grouped the cultured neural stem cells into normal group,ischemia model group,hypoxia group (hypoxia + ischemia model group),Lip2000 group (hypoxia+ ischemia model group + Lip2000 null transfection group,PTEN transfection group (hypoxia +ischemia model group+Ad5-PTEN transfection group),and PTEN interference group (hypoxia+ ischemia model group+Lip2000+PTENsiRNA interference group).We detected the proliferation of neural stem cells in the groups at different time points,and PTEN protein expression of neural stem cells in each group after PTEN transfection, and the effect on iconic protein on Akt-PI3K signaling pathway.Results (1 )Nestin identification of the neural stem cells was tested by immunofluorescence.We observed green bright and typical cell spheroids under fluorescence microscope;the clear structure could be seen within spheroids.(2)We determined the proliferation of neural stem cells at different time points by MTT.After 36 h of culture,the neural stem cells had obvious proliferation in the hypoxia group and Lip2000 group compared with the model group, the PTEN transfection group and PTEN interference group (P 0.05);(4)After PTEN plasmid transfection,there were no apparent changes in the total Akt and bad in the groups,but compared with the normal group,PI3K,p-Akt,and p-bad expressions in model group and PTEN gene transfection group reduced (P <0.05),with the more obvious changes in the PTEN transfection group.PI3K,p-Akt,and p-bad expressions in the hypoxia group, Lip2000 group and PTEN interference group increased (P < 0.05 ). Conclusion Hypoxia contributes to promoting the proliferation of neural stem cells,and the inhibition of PTEN on PI3K/Akt might be the key factor for blocked proliferation of adult neural stem cells.