Research Of G-CSF Protect Neural Stem Cells From Hypoxia Injury

Objective: In order to investigate the relationship between granulocyte colony-stimulating factor and Anti-apoptotic, neuroprotective, we use cobalt chloride to establish hypoxic model of neural stem cells in vitro, explore expressional level of histone deacetylase 2, hypoxia-inducible factor-1α.Methods: Used C17.2 neural stem cell line as the research object and incubated by Co Cl2 to mimic the hypoxia injury in vitro. Divided into three group:normal control group, hypoxia group, and treatment with G-CSF group, with MTT method to observe different concentrations of G-CSF for the survival of NSCs, each group NSCs proliferation activity, microscope to observe the morphological changes of NSCs, Western Blot to detect the expressional level of HDAC2, HIF-1α.Results: 1.The survival rate of neural stem cells in hypoxia :(1) 50、100umol/L Co Cl2 group increased cell viability at 12 h, has mild proliferation effect,Other time points cell survival rate were decreased(P<0.05).(2) Different concentrations of Co Cl2(50, 100, 200, 400, 800 umol/L) in 24 h, 48 h and 72 h all had a concentration-dependent decreased cell survival rate(P<0.05).(3)200 umol/L Co Cl2 resulted in a time-dependent decrease cell viability. 400,800umol/L Co Cl2 for cell viability at each time point was less than 40%.2.G-CSF intervented hypoxia neural stem cell survival:(1)Different concentrations of G-CSF(10,50,100ng/ml) at 12 h,24h,48 h,72h,cell survival rates were higher than 0ng/ml G-CSF+Co Cl2 group( P <0.05).(2)10ng/ml G-CSF group in cell viability 24 h NSCs are high compared with other concentrations and time points(P <0.05), was 88.7 ± 1.2%. 3.Western Blot results:(1) 200umol/L Co Cl2 hypoxic neural stem cell model:HDAC2 expression level at 12,24,48,72 h decreased than 0h(P <0.05), HIF-1α expression level at 12,24,48,72 h increased than 0h(P <0.05).(2) Under normoxic conditions G-CSF treatment of neural stem cells after 24 h HDAC2and HIF-1α expression level increased,(P <0.05).(3) In hypoxia, The expression of HDAC2 in hypoxia group and G-CSF intervention group were lower than normal group, hypoxia group was lower than that G-CSF group.The expression of HIF-1α in hypoxia group and G-CSF intervention group were higher than normal group, hypoxia group was higher than that G-CSF group.Conclusions: G-CSF could reduce the apoptosis of neural stem cells in hypoxia, G-CSF upregulated the expression of HDAC2 and HIF-1α in hypoxia. The latter might be one of the mechanisms former exert anti-apoptotic and neuroprotective effects, there was a correlation between HDAC2 and HIF-1α.