维生素E对帕金森病模型细胞的抗凋亡作用及其机制

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摘要目的: 探讨维生素E(VE)对1甲基4苯基吡啶离子(MPP+)诱导的神经元凋亡的保护作用及其可能机制。方法: 用重组鼠神经生长因子(nerve growth factor，NGF)对大鼠PC12细胞株进行诱导分化，加入500 μmol/L的MPP+制成帕金森病细胞凋亡模型，再给予不同浓度的VE处理细胞；Hoechst 33342染色后通过荧光显微镜观察各组细胞形态；应用流式细胞术观察各组细胞的凋亡率和坏死率；采用蛋白质印迹法检测各组细胞中细胞周期依赖蛋白激酶5（cyclindependent kinase 5，CDK5）、calpains的表达。结果：给予NGF后第2天可见PC12细胞突起数量增多、长度变长，细胞胞体变大、形态不规则；随着VE浓度升高，细胞凋亡率明显下降(P＜0.01)；VE浓度为200 μmol/L时细胞活性最高(P＜0.01)，细胞核固缩和碎裂现象最少见。随着VE浓度升高，PC12细胞内CKD5、calpains的表达逐渐下降。结论： VE通过抑制细胞凋亡通路中calpains通路，在帕金森病细胞模型中发挥神经元保护作用。

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关键词 ：维生素E, 帕金森病, 细胞凋亡, calpains

Abstract：Objective: To explore the protective role of Vitamin E(Vit E) in 1methyl4phenylpyridinium(MPP+)induced neuronal apoptosis and the probable mechanism. Methods: Nerve growth factor(NGF) was used to induce PC12 cells differentiating to neurons. MPP+ were added into culture medium to establish Parkinson′s disease(PD) cell model which then be added with different concentrations of Vit E to protect apoptotic neurons. Fluorescence microscope was used to detect the cellular morphology after Hoechst 33342 staining. The apoptotic and necrotic rate in each group was detected by FCM. Western blotting was used to detect the expression of CKD5 and calpains in each group PC12 cells. Results: Cells clustered and had good refractive power under an inverted microscope in the 2nd day after NGF added. Nuclear pyknosis and fragmentation was least in 200 μmol/L concentration of Vit E under fluorescence microscope. FCM showed cell viability elevated (P＜0.01) and apoptotic rate declined (P＜0.01) with increasing of concentration of Vit E. Cell viability reached the peak (P＜0.01) and apoptotic rate reduced to the bottom (P＜0.01) in 200 μmol/L concentration of Vit E. Expressions of CKD5 and calpains decreased with increasing concentration of Vit E. Conclusion: Vit E plays an important role in neuronal protection by inhibiting calpains pathways in PD.

收稿日期: 2016-09-22

基金资助:

镇江市社会发展项目（SH2015024；SH2015047）；江苏省“333工程”资助项目（BRA2014173）；江苏省“六大人才高峰”资助项目（WSN038）

通讯作者: 于明（通讯作者），主任医师，硕士生导师，E-mail:yuming7251@163.com

作者简介: 杨溢（1985—），男，江苏泰兴人，硕士，医师，主要从事老年神经退行性疾病的研究

引用本文:

杨溢，何悦，陈思远，于明. 维生素E对帕金森病模型细胞的抗凋亡作用及其机制[J]. 江苏大学学报:医学版, 2017, 27(01): 27-30,35. YANG Yi, HE Yue, CHEN Si-yuan, YU Ming.
Anti-apoptotic effects and potential mechanism of Vitamin E on cellular model of Parkinson′s disease. Journal of Jiangsu University(Medicine Edition), 2017, 27(01): 27-30,35.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2017/V27/I01/27

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