关键词: 氧/糖剥夺, 神经干细胞, 葡萄糖, 神经损伤与保护

Abstract:

Objective　 To explore the effects of glucose concentration on the viability and proliferation of hypoxic adult neural stem cells (ANSCs) after OGD.　Methods　The ANSCs cell line from adult Fisher 344 rats were cultured in serum-free medium and identified using Nestin staining. Anoxic cultured ANSCs (1% O2, 94%N2，and 5%CO2 for  6 h) were treated with different concentrations of D-glucose (7.5, 17.5, 27.75, 41.75, 83.75 mmol/L). A normoxic-normoglycemic control group was also employed. CCK-8 colorimetric method was used to determine the survival and proliferation of ANSCs, as well Hoechst 33342 immunofluorescent staining was used to detect the apoptosis of ANSCs. Mannitol was used as a control to exclude a possible effect of osmolality on cell viability．Results　Compared with those of the normoxic-normoglycemic control group, the O.D. at 450 nm of OGD group was higher significantly. Furthermore, there was a significant cell viability decrease in cultures exposed to 7.5 mmol/L, 41.75 mmol/L and 83.75 mmol/L glucose after hypoxia compared to that of control (P<0.05) . There was no difference between 27.75 mmol/L and 17.5 mmol/L glucose groups, under the exclusion of the influence of osmolarity on cell viability．The CCK-8 detection results were consistant with that of microscopic observation.　Conclusions　Mildly elevated glucose concentration after hypoxia may protect NSCs against hypoxia.

Key words: Oxygen glucose deprivation/reoxgenation（OGD/R）, Neural stem cells, D-glucose, Neuroprotection and injury