葡萄糖对神经干细胞缺氧性损伤保护作用的实验研究

PROTECTION OF GLUCOSE FOR HYPOXIC NSCs

目的探讨葡萄糖对神经干细胞缺氧性损伤的保护作用。方法分离培养小鼠胚胎11.5d神经干细胞。将三气培养箱的氧气浓度调至5%,以制备缺氧环境。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。通过计数克隆形成率和MTT法检测干细胞的存活和增殖情况,AnnexinⅤ-FITC/PI染色、激光扫描共聚焦显微镜观察和计算其凋亡率。结果缺氧前加入30 mmol/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,但仍低于常糖常氧正常对照组。缺氧后再加入葡萄糖时,其保护作用不明显。结论缺氧前给予足够浓度的葡萄糖对神经干细胞的缺氧性损伤具有一定的保护作用。

Objective To explore the protection of glucose for hypoxic neural stem cells（NSCs）.Methods The NSCs of E11.5d were isolated and cultured.Different concentrations of glucose were added to the nutritive medium before and after 5%O2 120h,and a normoxic-normoglycemic control group was employed.Counting cloning efficiency and MTT colorimetric method were used to detect the survival and proliferation of NSCs,and Annexin Ⅴ-FITC/PI immunofluorescent staining was used to detect apoptosis of NSCs by laser confocal scanning microscopy.Results Compared with those of the normoglycemic hypoxic group,the survival and proliferation of NSCs significantly intensified when 30mmol/L glucose was added before hypoxia,but were lower than those of the normoxic-normoglycemic control group.Glucose added after hypoxia had no obvious protection.Conclusion Sufficient glucose added before hypoxia can certainly protect NSCs agaist hypoxia.