基础医学与临床 ›› 2014, Vol. 34 ›› Issue (4): 480-484.

• 研究论文 • 上一篇    下一篇

过表达GBA诱导SH-SY5Y细胞自噬和降低α-synuclein的表达

范成河,王雪晶,楚广磊,马耀华,荆婧,滕军放   

  1. 郑州大学第一附属医院
  • 收稿日期:2013-07-18 修回日期:2013-10-26 出版日期:2014-04-05 发布日期:2014-03-31
  • 通讯作者: 滕军放 E-mail:13838210077@163.com
  • 基金资助:
    Numb/Notch信号通路调控一种新的蛋白修饰模式-

Overexpression of GBA induces autophagy and decreases the expression level of α-synuclein in SH-SY5Y cells

  • Received:2013-07-18 Revised:2013-10-26 Online:2014-04-05 Published:2014-03-31

摘要: 目的 探讨葡萄糖脑苷脂酶(GBA)对SH-SY5Y细胞自噬及α-突触核蛋白(α-synuclein)表达水平的影响。方法 将EGFP-N1、EGFP-GBA质粒转染SH-SY5Y细胞,Western blot检测Beclin 1 、LC3-II/LC3-I及α-synuclein蛋白的表达,荧光显微镜观察自噬泡的形成及LC3与GBA在细胞内的共定位。结果 与转染EGFP-N1组相比,转染EGFP-GBA组上调Beclin 1和LC3-II/LC3-I 蛋白的表达水平(P<0.05),而α-synuclein表达水平则下降(P<0.05);与转染EGFP-N1组相比,转染EGFP-GBA组自噬空泡数目明显增加,外源性LC3免疫荧光表达明显增强,且GBA与LC3存在共定位。结论 GBA上调自噬水平,降低α-synuclein蛋白的表达。

关键词: 葡萄糖脑苷脂酶, α-突触核蛋白, 自噬

Abstract: Objective To investigate the mechanism of autophagy induced by GBA and the regulation of GBA on the expression of α-synuclein in SH-SY5Y cells. Methods The recombinated plamid EGFP-N1, EGFP-GBA were transfected into SH-SY5Y cells through eukaryotic cell transfection technique. The expression of beclin1, LC3-II/LC3-I and α-synuclein in the level of protein was detected through Western blot,MDC staining and cellular immunofluorescence microscopy were performed to observe the change of autophagic vacuolization and their subcellular colocalization. Results Compared to EGFP-N1 group, the expression of Beclin1 and LC3-II/LC3-I in group of overexpression EGFP-GBA were significantly increased, and the average number of autophagic vacuoles per cell increased, however, the expression of α-synuclein was significantly decreased (P<0.05) . Besides, the signals for LC3, GBA and the extent of their co-localization increased in GBA groups. Conclusion GBA protein decreases the expression level of α-synuclein and induces autophagy in SH-SY5Y cells.

Key words: GBA, α-synuclein, autophagy

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