磷酸化 Smad3在神经元样细胞氧糖剥夺中的表达变化

摘要

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（Na2 S2 O4）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12 h 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 3 h 后细胞存活率显著下降，至 OGD 12 h 降至最低54．7％；细胞内磷酸化 Smad3表达在 OGD 3、6 h 与 OGD 0 h 组相比略升高，至 OGD 12 h 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.