Establishment of in vitro oxygen-glucose deprivation/reoxygenation axon injury model in rat hippocampal neurons / 第三军医大学学报

ABSTRACT

Objective To establish an in vitro oxygen-glucose deprivation/reoxygenation axon injury model in rat hippocampal neurons in order to found a basis for research concerning the injury of oxygen glucose deprivation and axons regeneration. MethodsThe hippocampal neurons isolated from rats 24 h after born and cultured for 7 d were exposed to D-hanks solution with nitrogen gas instead of the original culture medium for 0.5, 1.0 and 1.5 h respectively, and then continue to be cultured in the original culture medium with oxygen. LDH content in the culture media was measured at 1, 5, 24, 48 and 72 h after reoxygenation. The morphological changes of neuron and axon were observed with inverted phase contrast microscopy. ResultsAfter oxygen-glucose deprivation/reoxygenation injury, hippocampal neurons became darker, swollen, and were with shorten axons. With the elapse of time, the LDH content was increased. The survival rate of hippocampal neurons was higher and the change of axon length was more obvious in the group of oxygen-glucose deprivation for 0.5 h than in the other groups. ConclusionAn in vitro oxygen-glucose deprivation/reoxygenation axon injury model in rat hippocampal neurons is successfully established.