《中国康复理论与实践》

《中国康复理论与实践》

• 基础研究 • 上一篇    下一篇

丁苯酞对1-甲基-4-苯基吡啶离子诱导细胞自噬的影响①

贾玉凤 1，吴庆文 1，程月发 2，陈娟 1，孟奇 3   

  1. 1.华北理工大学护理与康复学院，河北唐山市 063000；2.华北理工大学冀唐学院，河北唐山市 063000；3. 四川大学华西公共卫生学院，四川成都市 610041。
  • 出版日期:2016-04-25 发布日期:2016-06-01

Effect of Butylphthalide on Autophagy of SH-SY5Y Cells Induced by l-methyl-4-phenyl-pyridiniumion

JIA Yu-feng1, WU Qing-wen1, CHENG Yue-fa2, CHEN Juan1, MENG Qi4   

  1. 1. College of Nursing and Rehabilitation of North China University of Science and Technology, Tangshan, Hebei 063000, China; 2. Jitang College of North China University of Science and Technology, Tangshan, Hebei 063000, China; 3. West China School of Public Health, Sichuan University, Chengdu, Sichuan 610041, China
  • Published:2016-04-25 Online:2016-06-01

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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，Westernblotting 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 mRNA 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mRNA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。

关键词: 丁苯酞, 帕金森病, SH-SY5Y细胞, 自噬, 大鼠

Abstract: Objective To observe the effects of butylphthalide on the expression of autophagy-related protein and mRNA in l-methyl-4-phenyl-pyridiniumion (MPP+)- induced SH-SY5Y cells, and to explore the protective effect and possible mechanism of butylphthalide to the cell model of Parkinson's disease. Methods The SH-SY5Y 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-SY5Y cells induced by MPP+ was measured with MTT assay, the morphology of SH-SY5Y cells was observed. The expression of microtubule associated protein 1 light chain 3 (LC3)-II/I and Beclin 1 protein was detected by Western blotting. And the expression of LC3-II/I and Beclin 1 mRNA 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-SY5Y cells induced by MPP+, which may affect Parkinson's disease by inducing autophagy.

Key words: butylphthalide, Parkinson's disease, SH-SY5Y cells, autophagy, rats