HT22海马神经元细胞氧糖剥夺再灌注模型的建立

【目的】探讨HT22海马神经元细胞氧糖剥夺再灌注(OGD/R)模型的建立条件。【方法】将HT22海马神经元细胞分为正常对照组和模型组,以不同的密度接种至96孔板,正常对照组细胞始终在正常气体条件下培养,模型组先采用缺氧小室对细胞进行缺氧处理8 h,然后给予不同时间的复氧处理。应用细胞计数试剂盒8(CCK-8)测定细胞活力,采用微板法检测乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)的活力及丙二醛(MDA)的含量,并进行细胞形态学观察。【结果】与正常对照组比较,接种密度9000、5000、3000个/孔的HT22细胞在复氧6 h时均明显损伤,细胞存活率降低,上清液中LDH活力及MDA含量升高,SOD活力下降,差异有统计学意义(P<0.05)。【结论】HT22细胞OGD/R模型的最佳造模条件是缺氧8 h复氧6 h。

Establishment of HT22 Cell Oxygen-glucose Deprivation Reperfusion Model

Objective To explore the establishment conditions of oxygen-glucose deprivation and reperfusion(OGD/R)model in HT22 hippocampus neurons.Methods The HT22 cells were divided into normal control group and model group.The HT22 cells were inoculated into 96-well plates at different densities.The HT22 cells in the normal control group were always cultured under normal gas conditions.The model group was treated with hypoxia chamber for 8 h,and then given reoxygenation treatment for different time.The cell viability was measured by cell counting kit 8(CCK-8).The activity of lactate dehydrogenase(LDH),superoxide dismutase(SOD)and the content of malondialdehyde(MDA)were detected by microplate method,and the cell morphological feature was observed.Results Compared with the normal control group,the HT22 cells with 9000,5000,3000 cells/well were significantly damaged after reoxygenation for 6 h,the cell viability was decreased,and the LDH activity and MDA content were increased and SOD activity was decreased in the supernatant,the difference being statistically significant(P<0.05).Conclusion The optimal modeling condition for OGD/R model of HT22 cells was hypoxia for 8 h and reoxygenation for 6 h.