基础医学与临床 ›› 2019, Vol. 39 ›› Issue (4): 495-500.

• 研究论文 • 上一篇    下一篇

重组人源galectin-1通过调节自噬水平保护神经元

李毅1,伍超1,任静1,高歌1,段春礼2,杨慧3,鲁玲玲4   

  1. 1. 首都医科大学
    2. 首都医科大学神经科学研究所
    3. 首都医科大学北京神经科学研究所 北京神经再生与修复研究重点实验室
    4. 首都医科大学北京神经科学研究所
  • 收稿日期:2018-09-25 修回日期:2018-12-21 出版日期:2019-04-05 发布日期:2019-03-26
  • 通讯作者: 鲁玲玲 E-mail:lllu@ccmu.edu.cn
  • 基金资助:
    国家重点研究发展计划;北京市教育委员会科技发展计划一般项目

Recombinant human galectin-1 protects neuron by regulating autophagy

  • Received:2018-09-25 Revised:2018-12-21 Online:2019-04-05 Published:2019-03-26
  • Contact: Ling-ling LU E-mail:lllu@ccmu.edu.cn

摘要: 目的 制备重组人源半乳糖凝集素1(rhGalectin-1),并研究其对受损神经元自噬水平的促进作用。方法 利用PCR技术构建pEGX-4T-1-Galectin-1原核生物蛋白表达质粒,构建工程菌。收集rhGalectin-1蛋白,并进行纯化及纯度鉴定。用细胞毒性物质鱼藤酮(rotenone)处理神经母细胞瘤SK-N-SH细胞系,检测在有无rhGalectin-1预处理的情况下自噬标志物LC3 II以及p62的水平,分析不同组别细胞的自噬水平。结果:成功制备了具有生物学活性的rhGalectin-1,发现rhGalectin-1可降低鱼藤酮对SK-N-SH细胞的毒性作用。鱼藤酮处理SK-N-SH细胞后,其自噬水平有所降低。表现为LC3 II水平的降低以及p62水平的增加(p<0.05)。而rhGalectin-1预处理组细胞自噬水平明显增加,LC3 II水平的升高,p62水平的降低(p<0.05)。当用自噬阻断剂bafilomycin阻断自噬体降解后,可观察到rhGalectin-1预处理并给予鱼藤酮刺激组较单独鱼藤酮刺激组LC3 II水平明显增加(p<0.05),此现象说明了rhGalectin-1处理的确增强了自噬水平而不是抑制自噬体的降解。结论:rhGalectin-1可以增强神经元的自噬水平,这一特性可能在神经元的毒性抵抗以及损伤后修复过程中发挥重要作用。

关键词: 关键词:半乳糖凝集素1, 蛋白纯化, 神经元, 鱼藤酮, 自噬

Abstract: Objective To prepare recombinant human Galectin-1(rhGalectin-1) and study its role in promoting autophagy of damaged neurons. Methods Plasmid of pEGX-4T-1-Galectin-1 was constructed by PCR and then transformed into E.coli BL21(DE3). SK-N-SH cells treated with rotenone and pretreated with or without rhGalectin-1. LC3 II and p62 levels were tested by Western blot.. Results Galectin-1 reduces the toxic effects of rotenone on SK-N-SH cells. After treatment of SK-N-SH cells with rotenone , the decrease in autophagy levels was observed by a decrease in LC3 II levels and an increase in p62 levels. In the Galectin-1 pretreatment group, the autophagy level increased significantly, the level of LC3 II increased and the level of p62 decreased. When the autophagy blocker bafilomycin was used, the level of LC3 II in the Galectin-1 pretreatment group increased, indicating that Galectin-1 promoted the autophagy level of injured SK-N-SH neurons. Conclusions rhGalectin-1 can promote the autophagy response of injured neurons, which may play an important role in the toxicity resistance process of neurons.

Key words: Key words: Galectin-1, protein purification, neuron, rotenone, Autophagy

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