磷酸化Smad3在神经元样细胞氧糖剥夺中的表达变化

Changes of Smads mRNA in nerve-like cells after OGD injury

目的探讨磷酸化Smad3蛋白在神经元样PC12细胞氧糖剥夺中的表达变化。方法利用鼠神经生长因子(NGF)诱导大鼠嗜铬细胞瘤细胞(PC12)向神经元样细胞转化,应用无糖DMEM培养液联合连二亚硫酸钠(Na2S2O4)构建氧糖剥夺模型(oxygen-glucose deprivation,OGD),体外模拟神经元缺血性损伤。MTT检测神经元样细胞OGD 0,3,6和12h存活率,Western blot检测OGD前后磷酸化Smad3蛋白表达。结果成功建立神经元样PC12细胞的OGD模型;OGD 3h后细胞存活率显著下降,至OGD 12h降至最低54.7%;细胞内磷酸化Smad3表达在OGD 3、6h与OGD 0h组相比略升高,至OGD 12h时明显降低。结论磷酸化Smad3参与在神经元样细胞OGD早期ActA/Smads通路的活化。

Objective To explore changes of phosphorylated Smad3（p-smad3）expression in neuron-like PC12 cells after oxygen-glucose deprivation（OGD）.Methods Rat nerve growth factor （NGF）was used to induce the differentia-tion of rattus PC12 pheochromocytoma cells to neuron-like cells.Na2S2O4 and glucose-free DMEM were introduced to perform an OGD model.The survival rate of cells suffered by OGD was examined by MTT assay.And the protein ex-pression of p-smad3 measured by Western blot.Results PC12 cells were successfully differentiated into neuron-like cells after NGF exposure.The survival rate of cells were dramatically decreased after 3h of OGD exposure,and reached to its lowest point at 12h.Compared with OGD 0h,expression of p-Smad3 was slightly increased after 3 or 6 h,and sig-nificantly decreased at 12h.Conclusion p-Smad3 took part in the activation of ActA/Smads signaling in response to early time of OGD treatment.