The Effects Of Interfering Olig2 On Proliferation,Differentiation And Apoptosis Of Hippocampal Neural Stem Cells Mediated By MK-801

ObjectiveMouse hippocampal neural stem cells(NSCs)were cultured in vitro to investigate the effects of interfering Olig2 on proliferation,differentiation and apoptosis of hippocampal NSCs mediated by MK-801,and to investigate the regulation of Olig2 on hippocampal neurogenesis after MK-801 intervention.MethodsNewborn C57BL/6 mice within 24 hours were selected to be extracted hippocampal tissue for culturing NSCs which was identified by immunofluorescence staining.NSCs were treated with 200 ?M MK-801 for 24 h to establish a cell injury model which was infected by Olig2-shRNA lentivirus.NSCs were divided into 4 groups:(1)MK-801+ lentiviral interference group [MK801+Olig2-shRNA group,(M+O group)],(2)MK-801+ lentivirus The empty vector group [MK-801+LV-EGFP group,(M+L group)],(3)the MK-801 group(M group),(4)the Blank control group(Blank).The expression of Olig2 was detected by Western blot to determine whether the Olig2-shRNA lentiviral vector was successfully constructed.Western-blot ? Real-time PCR and immunocytochemistry were used to investigate the expression of Nestin,Sox2,Ki67,S100? and NeuN,which was used to observe the proliferation and differentiation change of NSCs.Flow cytometry was used to detect the apoptosis of NSCs.ResultsWestern-blot results showed the expression of Olig2 was significantly decreased in the Olig2-shRNA1 group(P<0.05);compared with M group,the levels of Olig2 in the M+O group was significantly reduced(P<0.05)and the levels of NeuN and Sox2 in the M+O group were significantly higher(P<0.05),but there was no significant difference in the M+L group and Blank group(P>0.05);qRT-PCR results showed that compared with M group,the expression of Nestin and Ki67 in M+O group were significantly higher(P<0.05),and the expression of Nestin in the Blank group was higher too(P<0.05),there was no significant difference in the expression of Nestin between M+L group and M group(P>0.05).there was no significant difference in the expression of Ki67 in the M+L group and Blank group(P>0.05).and there was no significant difference in the expression of S100? in the M+O group?M+L group and Blank group(P>0.05);Immunofluorescence results show that compared with M group,the expression of Ki67 in M+O group were significantly higher(P<0.05),but this was no significant difference in the M+L group and Blank group(P>0.05),and there was no significant difference in the expression of S100? in the M+O group?M+L group and Blank group(P>0.05);The results of flow cytometry showed that the early apoptosis and total apoptotic rate of M+O group were lower than those of M group(P<0.05).There was no significant difference in late apoptosis rate.The early apoptosis and total apoptotic rate of M group were significantly higher than those of Blank group(P<0.05).Conclusion1.MK-801 has decreased proliferation and increased apoptosis after treatment of NSCs2.Low expression of Olig2 can promote the proliferation of NSCs after schizophrenia,promote the differentiation of NSCs into neurons,reduce apoptosis,and increase hippocampal neurogenesis.