食品科学 ›› 2010, Vol. 31 ›› Issue (19): 190-193.doi: 10.7506/spkx1002-6630-201019041

• 基础研究 • 上一篇    下一篇

应用细胞模型研究蜂胶醇提物的抗氧化活性

吴正双1,2,董 捷2,张红城2,赵亮亮3,高文宏1 ,*   

  1. 1.华南理工大学轻工与食品学院 2.中国农业科学院蜜蜂研究所 3.哈尔滨商业大学食品工程学院
  • 收稿日期:2010-06-01 出版日期:2010-10-15 发布日期:2010-12-29
  • 通讯作者: 高文宏 E-mail:zzhc@sohu.com
  • 基金资助:

    国家现代农业(蜂)产业技术体系建设专项(NYCYTX-43);农业部公益性行业(农业)科研专项(nyhyzx07-041)

Application of Cell Culture Model to Antioxidant Activity Analysis of an Ethanol Extract from Propolis

WU Zheng-shuang1,2,DONG Jie2,ZHANG Hong-cheng2,ZHAO Liang-liang3,GAO Wen-hong1,*   

  1. 1. College of Light Industry and Food, South China University of Technology, Guangzhou 510641, China;
    2. Bee Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100093, China;
    3. College of Food Engineering, Harbin University of Commerce, Harbin 150028, China
  • Received:2010-06-01 Online:2010-10-15 Published:2010-12-29
  • Contact: GAO Wen-hong E-mail:zzhc@sohu.com

摘要:

为了研究蜂胶醇提物在细胞内的抗氧化活性(CAA),本实验将2′, 7′- 二氯荧光二乙酸盐(DCFH-DA)荧光探针装载到人肝癌Hep G2 细胞内,然后被进入到细胞内的2, 2′- 偶氮二异丁基脒二盐酸盐(ABAP)产生的活性氧氧化成有荧光的二氯荧光(DCF);以荧光值为指标,使用多功能酶标仪和荧光倒置显微镜分析蜂胶醇提物清除活性氧的能力。结果表明:蜂胶醇提物具有明显的抗氧化活性,并呈现良好的剂量- 效应关系。当CAA 值达到50 时,蜂胶醇提物质量浓度仅0.14mg/mL;当蜂胶醇提物质量浓度为0.5mg/mL 时,细胞内的活性氧显著减少,有绿色荧光的细胞数几乎减少一半,清除率接近50%。蜂胶醇提物具有非常好的细胞内抗氧化能力。

关键词: 人肝癌Hep G2 细胞, 活性氧(ROS), 细胞内抗氧化

Abstract:

A cellular antioxidant activity (CAA) assay for quantifying the antioxidant activity of an ethanol extract from propolis (EEP) provided by Bee Research Institute, Chinese Academy of Agricultural Sciences has been developed. Dichlorofluoresceindiacetate (DCFH-DA) as a probe, which is easily oxidized into fluorescent dichlorofluorescein (DCF) by reactive oxygen species (ROS), was trapped within cells and the method was used to measure the ability of EEP to prevent the formation of DCF by 2,2′-azobis(2-amidinopropane) dihydrochloride (ABAP)-generated peroxyl radicals in human liver carcinoma HepG2 cells with multi-functional microplate reader and inverted fluorescence microscope. The decrease in cellular fluorescence intensity compared to the control cells indicates the antioxidant capacity of EEP. In addition, this bioactivity depended on dose. The concentration of EEP corresponding to CAA unit50 was only 0.14 mg/mL. A ROS clearance rate of 47.05% was obtained at an EEP concentration of 0.5 mg/mL.

Key words: human liver carcinoma HepG2 cells, reactive oxygen species, cellular antioxidant activity

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