缺氧性神经干细胞钙离子浓度变化和葡萄糖保护作用的研究

Study on [Ca^(2+)]_i concentration change and glucose protection of hypoxic NSCs

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。

To observe the change of Ca^2＋ concentration and explore the protection of glucose for hypoxic neural stem cell （NSCs） , the NSCs at El 1.5 were isolated and cultured by above-mentioned empirical method. Different concentrations of glucose were added to nutritive medium before and after 5% O2 culture for 120 h, at the same time, normoxic-nomloglycemic commom control group was set up. MTT colorimetric method was used to detect the survival and proliferation of NSCs, and [ Ca2 ＋ ] i of NSCs was measured by laser confocal scanning microscope. Compared with normoglycemie hypoxic group, the survival and proliferation of NSCs significantly intensified when 30 mmol/L glucose was added before hypoxia, and [ Ca^2＋]i concentration lowered significantly compared with hypoxia control group. Glucose added after hypoxia has not obvious protection. The present study indicate that sufficient glucose added before hypoxia can certainly protect NSCs agaist hypoxia through inhibiting [ Ca^2＋]i overload.