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为了从钙离子角度探讨葡萄糖对神经干细胞缺氧性损伤保护作用机制,本实验将三气培养箱的氧气浓度调至5%制备缺氧环境。分别在缺氧前后于无血清培养基中加入不同剂量的葡萄糖,同时设常氧常糖正常对照。通过MTT法检测干细胞的存活和增殖情况,用激光扫描共聚焦显微镜和Fluo-3荧光探针标记技术检测神经干细胞内钙离子浓度。结果显示,缺氧前加入30mmol/L葡萄糖,神经干细胞的存活率和增殖率较常糖缺氧组明显增高,其胞内钙离子浓度显著低于常糖缺氧组。缺氧后再加入葡萄糖时,其保护作用不明显。本实验提示:缺氧前给予足够浓度的葡萄糖可通过抑制胞内钙超载机制对神经干细胞损伤起到一定的保护作用。
In order to explore the protective effect of glucose on hypoxic injury of neural stem cells from the perspective of calcium ion, oxygen concentration in three-gas incubator was adjusted to 5% in this experiment to prepare hypoxic environment. Before and after hypoxia in serum-free medium were added different doses of glucose, while the normal hypoxia normal control. The survival and proliferation of stem cells were detected by MTT assay. The concentration of calcium in neural stem cells was detected by laser scanning confocal microscopy and Fluo-3 fluorescent probe labeling. The results showed that adding 30mmol / L glucose before hypoxia, the survival rate and proliferation rate of neural stem cells were significantly higher than that of hypoxia and hypoxia groups, and the intracellular calcium concentration was significantly lower than that of hypoxia group. Hypoxia and then added glucose, its protective effect is not obvious. This experiment suggests that: given a sufficient concentration of glucose before hypoxia can inhibit neural stem cell damage by inhibiting intracellular calcium overload mechanism.