大鼠皮质星形胶质细胞氧糖剥夺后Slit2、Robo1的表达研究

Expression of Slit2 and Robo1 in astrocytes of rat cortex with oxygen-glucose deprivation

目的探讨神经生长导向因子slit2及其受体robo1在氧糖剥夺(oxygen-glucose deprivation,OGD)不同时间后大鼠皮质星形胶质细胞的表达变化及其意义。方法建立培养乳大鼠皮质星形胶质细胞OGD模型,并分为OGD 0h组(对照组)、2h组、4h组、6h组和12h组。相差显微镜观察各组的细胞形态,MTT法检测各组的细胞活力,RT-PCR法检测各组slit2mRNA和robo1 mRNA的相对表达情况。结果 (1)对照组、OGD 2h组、4h组、6h组和12h组的细胞活力(OD值)分别为0.756±0.013、0.681±0.016、0.563±0.033、0.404±0.024和0.222±0.026,细胞活力随着氧糖剥夺时间延长而下降,呈时间依赖性(P<0.01)。(2)氧糖剥夺后,统计结果显示不同组别星形胶质细胞slit2 mRNA和robo1 mRNA表达先逐渐升高,6h组达高峰(与对照组比,P<0.05),12h组明显下降(与6h组比,P<0.01)。结论 slit2/robo1信号通路可能参与缺血后抑制性微环境的形成,影响神经的再生修复过程。

Objective To investigate the expression patterns of the nerve growth guidance cue slit2 and its receptor Robo1 in rat cortex astrocytes with different severity of oxygen-glucose deprivation（OGD）.Methods Cultured cortex astrocytes were designed into five groups：OGD 0h group（control group）,2h group,4h group,6h group and 12h group.Morphological changes were observed by phase-contrast microscopy.MTT analysis was used to detect the cell viability.Reverse transcriptase-polymerase chain reaction（RT-PCR）was contributed to detect slit2 and Robo1 mRNA expressions.Results The cell activities（OD values）were 0.756±0.013,0.681±0.016,0.563±0.033,0.404±0.024 and 0.222±0.026 in OGD 0h group,2h group,4h group,6h group and 12h group respectively（P〈0.01）,with time-dependent manner.After oxygen-glucose deprivation,statistics showed that expressions of slit2 and Robo1 mRNA in astrocytes were first up-regulated,peaked at 6h（P〈0.05,compared with control group）,and then decreased.Compared with 6h group,both of the expressions were decreased significantly in 12h group（both P〈0.01）.Conclusion The expression of slit2/Robo1 in astrocytes can be up-regulated after OGD,suggesting that slit2/Robo1 may be involved in the regulation of neural regeneration.