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目的探讨不同浓度的葡萄糖对氧/糖剥夺条件下体外成年大鼠神经干细胞活力和增殖的影响。方法无血清培养成年Fisher 344大鼠海马神经干细胞系,用Nestin和DAPI免疫荧光双染以确认其生物学特性。将三气培养箱氧气浓度调至1%以制备缺氧环境,将培养基换为无糖Earle’s平衡盐溶液缺氧培养6 h后更换为含有不同浓度葡萄糖(7.5、17.5、27.75、41.75、83.75 mmol/L)的培养基恢复正常条件继续培养后,检测神经干细胞的活力和形态学改变。同时设置常氧常糖为正常对照组和甘露醇为渗透压对照组。结果 CCK-8检测显示,常糖常氧正常对照组细胞吸光度值较缺氧模型组高,7.5 mmol/l糖培养组以及41.75 mmol/L和83.75 mmol/L糖培养组吸光度值均较正常糖缺氧组(17.5 mmol/L)低,差异有统计学意义(P<0.05);而27.75 mmol/L糖培养组吸光度值较正常糖缺氧组低,差异无统计学意义,且排除了高糖培养基引起的渗透压力的影响。各组细胞形态学改变与CCK-8检测结果一致。结论缺氧后轻度升高的糖对体外缺氧成年神经干细胞的损伤可能有保护作用,而较低浓度和更高浓度的葡萄糖则加重其损伤。
Objective To investigate the effects of different concentrations of glucose on the activity and proliferation of neural stem cells in adult rats in vitro under oxygen / glucose deprivation. Methods The adult rat hippocampal neural stem cell line of Fisher 344 was cultured in serum-free medium. Double immunofluorescence staining with Nestin and DAPI was used to confirm the biological characteristics. The three-gas incubator oxygen concentration adjusted to 1% to prepare hypoxic environment, the medium was replaced with sugar-free Earle’s Balanced Salt Solution 6 h after hypoxia and replaced with different concentrations of glucose (7.5,17.5,27.75,41.75,83.75 mmol / L) medium to resume normal conditions continue to culture, detection of neural stem cell activity and morphological changes. At the same time set the normoxia sugar as normal control group and mannitol as osmotic pressure control group. Results The CCK-8 assay showed that the absorbance values of normal sugar and normoxia control group were higher than those of hypoxia model group, and the absorbance values of sugar culture group of 7.5 mmol / l, 41.75 mmol / L and 83.75 mmol / L were higher than that of normal sugar (17.5 mmol / L), the difference was statistically significant (P <0.05); while the absorbance value of 27.75 mmol / L glucose culture group was lower than normal glucose hypoxia group, the difference was not statistically significant, and excluded the high Effect of sugar medium on osmotic pressure. Cell morphological changes in each group and CCK-8 test results. Conclusions: Mild hypoxia increased the damage of hypoxic adult neural stem cells in vitro. However, lower concentrations and higher concentrations of glucose aggravate the injury.