【摘要】 目的探讨Smads mRNA在神经元样PC12细胞氧糖剥夺中的表达变化。方法鼠神经生长因子(NGF,50ng/ml)诱导大鼠嗜铬细胞瘤PC12细胞的神经元样转化,微管相关蛋白(Microtubule-associated protein 2,MAP2)染色,共聚焦显微镜验证其神经元特性。利用连二亚硫酸钠(Na2S2O4,1mmol/L)构建氧糖剥夺模型(oxygen-glucosedeprivation,OGD),模拟神经细胞缺血性损伤。实时定量RT-PCR检测神经元样细胞OGD 0,3和6h后Smad2、4、7基因mRNA的表达变化。结果成功诱导神经元样PC12细胞。OGD处理3h后细胞Smad2、4、7基因mRNA的表达与OGD0h组相比明显升高,分别超过17.7%,413.4%,75.2%。OGD6h,Smad2、7基因mRNA的表达较OGD0h组下调80.9%,57.4%,Smad4mRNA表达较OGD3h组下调73.0%,但与OGD0h相比,表达提高38.8%。结论 ActA/Smads通路激活发生在神经元样细胞OGD损伤的早期(OGD3h);Smad7转录水平上参与该通路的负性调节过程。更多还原

【Abstract】 Objective To explore changes of Smads mRNA expression in nervelike PC12 cells after oxygen-glucose deprivation.Methods Nerve growth factor(NGF,50ng/ml) was used to induce the differentiation of rattus PC12 pheochromocytoma cells to nerve-like cells.After Microtubule-associated protein 2 immunostaining,nerve-like PC12 cells were observed under the confocal microscopy to identify its neuronal characteristic.Oxygen-glucose deprivation(OGD) model was introduced to those nerve-like PC12 cells by hyposulfite of soda,to imitate the pathological process that nerve cells suffered during the ischemic stroke.The transcription of Smad2,4,7 mRNA in nerve-like PC12 cells was detected by Real-Time RT-PCR,after 0,3 or 6h of OGD treatment.Results After 5 days of NGF exposure,PC12 cells were successfully differentiated into nerve-like cells with MAP2 highly expression.The mRNA level of Smad2、4、7 genes in OGD3h group increased over 17.7%,413.4% and 75.2% respectively,compared to that in OGD0h group.After extending the exposure time of OGD to 6h,the mRNA expression of Smad2、7 were significantly decreased by 80.9% and 57.4%,compared to that in OGD0h group.Smad4 mRNA increased over 38.8% compared to OGD0h group,but decreased more than 73.0% compared to OGD3h group.Conclusion ActA/Smads pathway was activated in the early stage of OGD treatment(OGD3h),and the mRNA expression of Smad7 participated in the negative regulation of this pathway.更多还原