氧糖剥夺离体神经元缺血模型的建立

Development of neuronal ischemic model by oxygen and glucose deprivation in vitro

目的：探讨建立一种方便、实用的神经细胞离体氧、糖剥夺体外“缺血”模型。 方法：新生大鼠皮层－基底节神经元原代分散培养14天后，将培养液分别换为含葡萄糖和不含葡萄糖的Earle氏液。不含葡萄糖组置入一个经改造的保鲜盒内（体积2000ml），密闭并通入混合气体（含95% N2，5% CO2）4h，采用LDH和MTT法检测神经元的损害。结果：氧、糖剥夺（ OGD）4h组随着复氧时间的延长，神经元的活性逐渐下降，培养液中LDH释放量逐渐增加，与对照组比较有显著性差异。结论：建立了方便、实用的体外神经元“缺血”模型，为进一步研究缺血性神经元损害机制及有关药物的保护作用提供了一手段。

Objective: To make a convenient and practical in vitro ischemia model by oxygen and glucose deprivation(OGD) in cultured neuronal cells. Methods: The cultured neurons dissociated from the cortices and striata of the neonatal rats were observed on the 14th day after culture. The culture media were exchanged by Earle's solution with glucose or without glucose, respectively. The group without glucose was placed in a modified chamber(volume, 2000ml) and mixture gas (95% N 2, 5% CO 2) was put inside. The neuronal injury was detected by the lactate dehydrogenase (LDH) release and the tetrazolium salt 3-(4,5-dimethyl-2-thiazolyl)-2,5 diphenyl-2H tetrazolium bromide (MTT) assay after OGD. Results: The activity of neurons (MTT assay) was decreased and the amount of LDH release from damaged neurons was increased significantly in OGD group than in control (Con) group after reoxygen. Conclusion: Our results suggested that in vitro ischemia model may be induced easily by OGD in cultured neurons, and may be used to further study the mechanism of ischemic neurounal cell injury and its protection.