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从富硒螺旋藻中纯化得到的含硒别藻蓝蛋白通过抑制 ROS 生成来减轻 AAPH 诱导的人红细胞氧化应激。

Selenium-containing allophycocyanin purified from selenium-enriched Spirulina platensis attenuates AAPH-induced oxidative stress in human erythrocytes through inhibition of ROS generation.

机构信息

Department of Chemistry, Jinan University, Guangzhou, Guangdong Province, China.

出版信息

J Agric Food Chem. 2011 Aug 24;59(16):8683-90. doi: 10.1021/jf2019769. Epub 2011 Aug 1.

Abstract

Both selenium and allophycocyanin (APC) have been reported to show novel antioxidant activities. In this study, a fast protein liquid chromatographic method for purification of selenium-containing allophycocyanin (Se-APC) from selenium-enriched Spirulina platensis and the protective effect of Se-APC on 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress have been described. After fractionation by ammonium sulfate precipitation, and separation by DEAE-Sepharose ion-exchange and Sephacryl S-300 size exclusion chromatography, Se-APC with purity ratio (A652/A280) of 5.30 and Se concentration of 343.02 μg g(-1) protein was obtained. Se-APC exhibited stronger antioxidant activity than APC by scavenging ABTS (2,2'-azinobis-3-ethylbenzothiazolin-6-sulfonic acid) and AAPH free radicals. The oxidative hemolysis and morphological changes induced by AAPH in human erythrocytes were effectively reversed by coincubation with Se-APC. Lipid oxidation induced by the pro-oxidant agent cupric chloride in human plasma, as evaluated by formation of conjugated diene, was blocked by Se-APC. The accumulation of malondialdehyde, loss of reduced glutathione, and increase in enzyme activities of glutathione peroxidase and reductase induced by AAPH in human erythrocytes were effectively suppressed by Se-APC. Furthermore, Se-APC significantly prevented AAPH-induced intracellular reactive oxygen species (ROS) generation. Taken together, our results suggest that Se-APC demonstrates application potential in treatment of diseases in which excess production of ROS acts as a casual or contributory factor.

摘要

硒和别藻蓝蛋白(APC)都已被报道具有新型抗氧化活性。本研究描述了从富硒螺旋藻中快速分离含硒别藻蓝蛋白(Se-APC)的快速蛋白质液相色谱法,以及 Se-APC 对 2,2'-偶氮双(2-脒基丙烷)二盐酸盐(AAPH)诱导的氧化应激的保护作用。经硫酸铵沉淀分级、DEAE-琼脂糖离子交换和 Sephacryl S-300 分子筛层析分离后,获得纯度比(A652/A280)为 5.30 和硒浓度为 343.02μg g(-1) 蛋白的 Se-APC。Se-APC 通过清除 ABTS(2,2'-偶氮双-3-乙基苯并噻唑啉-6-磺酸)和 AAPH 自由基,表现出比 APC 更强的抗氧化活性。Se-APC 可有效逆转 AAPH 诱导的人红细胞氧化溶血和形态变化。通过形成共轭二烯评估的铜离子引发剂氯化铜在人血浆中诱导的脂质氧化被 Se-APC 阻断。Se-APC 有效抑制 AAPH 诱导的人红细胞丙二醛积累、还原型谷胱甘肽损失以及谷胱甘肽过氧化物酶和还原酶活性升高。此外,Se-APC 可显著防止 AAPH 诱导的细胞内活性氧(ROS)生成。总之,我们的结果表明,Se-APC 在治疗以过量 ROS 为因果或促成因素的疾病方面具有应用潜力。

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