食品科学 ›› 2010, Vol. 31 ›› Issue (7): 81-85.doi: 10.7506/spkx1002-6300-201007018

• 基础研究 • 上一篇    下一篇

壳寡糖对自由基的清除及对N9小胶质细胞的保护作用

张 吉1,刘洪涛2,李秀英1,谢晓艳1,李文明1,于 超1 ,*   

  1. 1.重庆医科大学生命科学研究院 2.中国科学院大连化学物理研究所
  • 收稿日期:2009-07-20 出版日期:2010-04-01 发布日期:2010-12-29
  • 通讯作者: 于超 E-mail:yuchaom@163.com
  • 基金资助:

    重庆市教育委员会科学技术研究项目(KJ070304);重庆医科大学人才启动基金资助项目(0200100263)

Chitooligosaccharide: Free Radicals Scavenging Activity and Protective Effect against Lipopolysaccharide-induced Oxidative Injury in Microglial Cell Line N9

ZHANG Ji1,LIU Hong-tao2,LI Xiu-ying1,XIE Xiao-yan1,LI Wen-ming1,YU Chao1,*   

  1. 1. Institute of Life Sciences, Chongqing Medical University, Chongqing 400016, China;
    2. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
  • Received:2009-07-20 Online:2010-04-01 Published:2010-12-29
  • Contact: YU Chao E-mail:yuchaom@163.com

摘要:

目的:研究壳寡糖(COS)的抗氧化作用及其对脂多糖(LPS)损伤N9 小胶质细胞的保护作用。方法:首先在细胞外检测COS 对DPPH 自由基、H2O2 和羟自由基的清除作用,然后采用LPS 诱导建立体外培养N9 小胶质细胞活化模型,将细胞分为正常对照组、LPS 模型组、LPS+COS 给药组(COS 质量浓度分别为50、100、200、300、400μg/mL)。观察各组细胞的形态;检测各组细胞培养液中一氧化氮(NO)含量;用流式细胞仪检测细胞内活性氧(ROS)水平并观察其荧光图像;用DNA 损伤实验观察各组细胞凋亡情况。结果:COS 能有效清除DPPH 自由基和羟自由基,对H2O2 无清除能力;COS 能减少活化的N9 细胞数量并明显降低LPS 活化的N9 细胞培养液中NO 水平,降低细胞内ROS 水平,减少LPS 诱导的细胞凋亡,保护N9 小胶质细胞。结论:COS 在N9 小胶质细胞内外均具有较强的抗氧化能力,能保护N9 小胶质细胞,减轻N9 小胶质细胞的氧化损伤。

关键词: 抗氧化, COS, 小胶质细胞, 细胞凋亡

Abstract:

Objective: To study anti-oxidant activity of chitooligosaccharide (COS) and its protective effect against lipopolysaccharide (LPS)-induced oxidative injury in microglial cell line N9. Methods: The antioxidant activity of COS in vitro was evaluated by DPPH and hydroxyl free radical and hydrogen peroxide scavenging assays. Microglial cell line N9 was treated with LPS alone at the dose of 1 μg/mL or co-treated with LPS at the dose of 1 μg/mL and COS at doses of 50, 100, 200, 300 μg/mL and 400 μg/mL, respectively. Cell morphology was examined under microscope with HE staining and NO level in cell culture supernatant was determined. Intracellular generation of reactive oxygen species (ROS) was detected using 2',5' dichlorofluorescin diacetate (DCFH-DA) and flow cytometry combined with fluorescence microscope. Cell apoptosis was detected by DNA ladder assay. Results: COS exhibited an effective scavenging activity against DPPH and hydroxyl free radicals but no scavenging activity against H2O2. Meanwhile, COS attenuated the activation of microglial cell line N9 induced by LPS. The level of NO in cell culture supernatant was significantly decreased due to the presence of COS, and ROS generation induced by LPS was inhibited by COS in a dose-dependent manner. COS also inhibited the apoptosis of microglial cell line N9 induced by LPS. Conclusion: The antioxidant activity of COS is beneficial for protecting microglial cell line N9 from oxidative injury induced by LPS.

Key words: anti-oxidant activity, chitooligosaccharide, microglial cell, apoptosis

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