摘要
目的研究丁苯酞对1-甲基-4-苯基吡啶离子(MPP+)诱导的SH-SY5Y细胞自噬相关因子的影响,探讨丁苯酞对帕金森病细胞模型的神经保护作用机制。方法将SH-SY5Y细胞分为空白对照组(A组)、MPP+组(B组)、雷帕霉素预处理+MPP+组(C组)和丁苯酞预处理+MPP+组(D组)。采用噻唑蓝(MTT)比色法测定细胞相对存活率,倒置相差显微镜观察细胞形态的变化,Western blotting检测微管相关蛋白1轻链3(LC3)-Ⅱ/Ⅰ、Beclin 1蛋白表达,RT-PCR检测LC3-Ⅱ/Ⅰ和Beclin 1 m RNA表达。结果 B组SH-SY5Y细胞存活率显著低于A组(t=20.270,P<0.001),C组和D组显著高于B组(t>8.770,P<0.001),且C组和D组间无显著性差异(t=2.270,P=0.064)。与A组相比,B组LC3-Ⅱ/Ⅰ、Beclin 1蛋白和m RNA表达明显增加(t>6.647,P<0.01);C组和D组明显高于B组(t>3.630,P<0.01),C组与D组间无显著性差异(t<2.238,P≥0.05)。结论丁苯酞可以提高MPP+诱导的SH-SY5Y损伤细胞存活率,其可能通过诱导帕金森病模型细胞自噬,发挥治疗作用。
Objective To observe the effects of butylphthalide on the expression of autophagy- related protein and m RNA in l- methyl-4-phenyl-pyridiniumion（MPP＋）- induced SH-SY5 Y cells, and to explore the protective effect and possible mechanism of butylphthalide to the cell model of Parkinson＇s disease. Methods The SH-SY5 Y cells were divided into control group（A）, MPP＋group（B）, rapamycin pretreated＋ MPP＋group（C） and Butylphthalide pretreated ＋ MPP＋group（D）. The relative viability of SH-SY5 Y cells induced by MPP＋was measured with MTT assay, the morphology of SH-SY5 Y cells was observed. The expression of microtubule associated protein 1 light chain3（LC3）-II/I and Beclin 1 protein was detected by Western blotting. And the expression of LC3-II/I and Beclin 1 m RNA were assayed by real- time quantitative reverse transcription- PCR（RT- PCR）. Results The viability rates of cells were significantly lower in group B than in group A（t=20.270, P〈0.001）, and were significantly higher in groups C and D than in group B（t〉8.770, P〈0.001）, however, there was no significantly difference between groups C and D（t=2.270, P=0.064）. The expression of LC3-II/I and Beclin 1 was higher in group B than in group A（t〈6.647, P〈0.01）, and was higher in groups C and D than in group B（t〉3.630, P〈0.01）, however, there was no significantly difference between groups C and D（t〈2.238, P≥0.05）. Conclusion Butylphthalide could prevent the injury of SH-SY5 Y cells induced by MPP＋,which may affect Parkinson＇s disease by inducing autophagy.
出处
《中国康复理论与实践》
CSCD
北大核心
2016年第4期422-427,共6页
Chinese Journal of Rehabilitation Theory and Practice
基金
河北联合大学博士科研启动基金项目(No.35647699)
