【摘要】 目的研究不同氧糖剥夺(OGD)时间和不同介质对N9小胶质细胞活力的影响,以建立体外小胶质细胞的OGD模型。方法以平衡盐溶液(BSS)和无糖DMEM作为OGD介质,OGD 10、30和60 min并复氧复糖24 h后,倒置相差显微镜下观察N9小胶质细胞形态变化,四氮唑盐复合物(XTT)比色法检测细胞活力变化,比较分析不同OGD持续时间和不同介质产生的结果差异。结果氧糖剥夺/复氧复糖后,各OGD处理组不规则细胞比例增多,部分细胞发生皱缩,细胞轮廓不清,折光度较差,细胞密度减小。各BSS介质的OGD处理组与对照组相比,N9小胶质细胞的活力产生不同程度降低,差别均有统计学意义(P<0.01)。各OGD处理组相比,OGD 30 min组和OGD 60 min组细胞活力低于OGD 10 min组,差别有统计学意义(P<0.01)。各无糖DMEM介质的OGD处理组与对照组相比,N9小胶质细胞的活力产生不同程度降低,差别均有统计学意义(P<0.01),且随着OGD处理时间的延长,细胞活力逐渐减小,各OGD处理组间细胞活力差别有统计学意义(P<0.05)。OGD 10 min处理后,无糖DMEM介质组细胞活力比BSS介质组细胞活力高(P<0.01),OGD 30 min处理后,无糖DMEM介质组细胞活力仍比BSS介质组细胞活力高(P<0.05),OGD 60 min处理后,2组间差别无统计学意义(P>0.05)。结论 OGD处理后,N9小胶质细胞形态发生改变,细胞活力与OGD持续时间呈负相关,体外小胶质细胞的OGD模型制备成功。模型中,2种OGD介质对细胞活力影响的差别随着OGD处理时间的延长逐渐缩小并最终消失。更多还原

【Abstract】 Objective To investigate the effects of different oxygen-glucose deprivation(OGD) durations and media on the cell viability of N9 microglia and establish an OGD model of cultured microglia in vitro.Methods Balanced salt solution(BSS) and glucose-free DMEM were used as OGD media.After OGD for 10,30 and 60 minutes and restoration for 24 hours,morphologic changes of N9 microglia were observed by inverted phase contrast microscope and the cell viability was measured by means of XTT colorimetric assay.Finally,results were analyzed and compared.Results After OGD and restoration,irregular cells of all OGD groups increased and some cells shrank.The outlines of some cells became obscure and the refractive index declined.The cell density of the OGD groups decreased.Compared with control group,the cell viability of all the OGD groups with BSS as medium decreased in varying degrees(P<0.01).The cell viability in OGD 30 minutes group and OGD 60 minutes group was lower than that in OGD 10 minutes group,and there was a statistically significant difference(P<0.01).Compared with control group,the cell viability in all the OGD groups with glucose-free DMEM as medium also decreased in varying degrees(P<0.01).Furthermore,Along with the extending of time,the cell viability gradually decreased.The difference in the cell viability between all OGD groups was statistically significant(P<0.05).After OGD for 10 minutes,the cell viability in glucose-free DMEM groups was higher than that in BSS groups(P<0.01).After OGD for 30 minutes,the cell viability in glucose-free DMEM groups still remained higher than that in BSS groups(P<0.05).But after OGD for 60 minutes,the cell viability of the two groups had no significant difference(P>0.05).Conclusion After OGD,the morphology of N9 microglia changed and there is a negative correlation between the cell viability and the OGD duration.The OGD model of microglia in vitro is successfully established.And in the model,the difference in the effects of the two OGD media on the cell viability narrowed gradually and disappeared as time extended.更多还原