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[谷胱甘肽与谷胱甘肽检测]

[Glutathione and glutathione assays].

作者信息

Rousar Tomás, Cervinková Zuzana, Muzáková Vladimíra, Kucera Otto, Lotková Halka, Krivákovaá Pavla

机构信息

Univerzita Karlova v Praze, Lékarská fakulta v Hradci Králové, Ustav fyziologie.

出版信息

Acta Medica (Hradec Kralove) Suppl. 2005;48(1):15-20.

Abstract

Glutathione, the very important intracellular antioxidant, is present in intracelullar environment in milimolar concentrations. Glutathione is a tripeptide molecule, which plays an essential role in the antioxidant system, as well as in maintenance of the intracellular redox state. This thiol compound exists in two forms, the reduced (GSH) and the oxidized (GSSG), and the ratio of both forms is crucial for the characterization of the oxidative stress in cells. Number of analytical methods have been developed for the measurement of the glutathione. Especially, High Performance Liquid Chromatography methods (HPLC) are mostly used linked to different types of detection, including electrochemical, UV/VIS or fluorimetric detection. Another approach for glutathione assay is using the spectral methods, either fluorimetric or spectrophotometric assays. In enzymatic assay, glutathione reductase reduces GSSG with simultaneous oxidation of specific substrate, which is sequentially photometrically detected. The fluorimetric method is based on the detection of derivatized GSH molecule.

摘要

谷胱甘肽是非常重要的细胞内抗氧化剂,在细胞内环境中以毫摩尔浓度存在。谷胱甘肽是一种三肽分子,在抗氧化系统以及维持细胞内氧化还原状态中起着至关重要的作用。这种硫醇化合物以两种形式存在,即还原型(GSH)和氧化型(GSSG),两种形式的比例对于表征细胞内的氧化应激至关重要。已经开发了多种分析方法来测量谷胱甘肽。特别是,高效液相色谱法(HPLC)大多与不同类型的检测方法联用,包括电化学检测、紫外/可见检测或荧光检测。另一种谷胱甘肽检测方法是使用光谱法,即荧光或分光光度法检测。在酶促检测中,谷胱甘肽还原酶将GSSG还原,同时特定底物被氧化,随后通过光度法进行检测。荧光法基于对衍生化的GSH分子的检测。

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