Seleno-polymannuronate Synthesis and Resistance to Oxidation and Apoptosis in Alzheimer's Disease Cells
Seleno-polymannuronate Synthesis and Resistance to Oxidation and Apoptosis in Alzheimer's Disease Cells
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阿尔茨海默氏病细胞中硒代聚甘露糖醛酸的合成以及抗氧化和凋亡
DOI：
10.7503/cjcu20120462
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发表时间：
2013-01-10
影响因子：
1
通讯作者：
Song Yun
中科院分区：
文献类型：
--
作者：
Zhu Zhi-Jie;Liu Qiong;Song Yun
缺硒与多种疾病密切相关。补充适量的硒对人体健康非常重要。由于不同的硒化合物具有不同的生物学效应，新产品的制备和功能研究是发现含硒药物的关键。在这项工作中，。以聚甘露醇(PM)为原料，合成了磺化聚甘露醇(S-PM)。然后用亚硒酸钠取代S-PM中的磺化基团制备了硒-聚甘露酸单酯(Se-PM)。以N2a-APP695-Sw细胞为AD模型，用纯化的Se-PM研究其抗氧化性能及对阿尔茨海默病(AD)的影响。CCK-8比色法检测细胞活力。用流式细胞仪检测细胞内活性氧(ROS)。激光扫描共聚焦显微镜检测线粒体膜电位和细胞色素C释放。Western Blot分析细胞凋亡和AD病理相关蛋白的表达水平。结果表明，Se-PM的最佳硒浓度为2.5mU/L，可显著提高细胞存活率。该浓度的Se-PM可保护细胞免受过氧化氢的氧化损伤，抑制细胞内ROS，提高超氧化物歧化酶和谷胱甘肽过氧化物酶活性，促进线粒体膜电位，抑制细胞色素C释放到细胞质。同时，Se-PM还能增加Bcl2的表达，降低Bar的表达，抑制AD病理相关蛋白BACE1和APP的表达。这些结果表明，Se-PM可通过抑制细胞凋亡和淀粉样斑块的形成来对抗AD，这是AD的一个重要病理特征。为抗AD新药的开发提供了基础数据。
Selenium deficiency is closely relate to multiple diseases. Supplementation with adequate amount of selenium is very important for human health. As various selenium compounds have different biological effects, preparation and functional study of new product are essential for the discovery of selenium-containing drug. In this work,. polymannuronate(PM) was used as the raw material to synthesize sulfonated polymannuronate( S-PM). Seleno-polymannuronate(Se-PM) was then prepared by the replacement of sulfonated group in S-PM with sodium selenite. The yield of Se-PM synthesis was 54% with a selenium content of 437.25 mu g/g. Purified Se-PM was used to study its antioxidative property and effect on Alzheimer's disease (AD), using N2a-APP695-sw cells as an AD model. Cell viability was detected by CCK-8 assay. Intracellular reactive oxygen species(ROS) was measured by flow cytometry. Mitochondrial membrane potential and cytochrome C release were detected by laser scanning confocal microscope. The expression levels of cell apoptosis and AD pathology relevant proteins were analyzed by Western Blot. The results showed that the optimum selenium concentration was 2. 5 mu mol/L for Se-PM to significantly increase cell viability. Se-PM at this concentration could prevent cells from the oxidative damage of hydrogen peroxide, inhibit intracellular ROS, increase the activity of superoxide dismutase and glutathione peroxidase, promote mitochonclrial membrane potential, and suppress cytochrome C release into cytoplasm. Meanwhile, Se-PM could also increase Bcl-2 expression and decrease Bar expression, inhibit the expression of AD pathology relevant proteins BACE1 and APP. Those results indicated that Se-PM could resist AD through the prevention of cell apoptosis and amyloid plaque formation, a key pathological feature of AD. It also provides basic data for the development of new anti-AD drug.