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富硒蛹虫草中一种新型硒多糖的结构表征及抗肿瘤活性

Structural characterization and antitumor activity of a novel Se-polysaccharide from selenium-enriched Cordyceps gunnii.

机构信息

State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, PR China.

出版信息

Food Funct. 2018 May 23;9(5):2744-2754. doi: 10.1039/c8fo00027a.

Abstract

Selenium (Se) has been recognized as an essential element. Animals and humans absorb and retain more organic Se than inorganic Se. Bio-transformation is a major approach to transform inorganic Se to organic Se. Cordyceps gunnii (C. gunnii), well known as a Chinese edible and medicinal fungus, has a variety of biological functions. C. gunnii and selenium were combined through liquid-fermentation to obtain selenium-enriched C. gunnii. A novel Se-polysaccharide (SeCPS-II) was extracted from selenium-enriched C. gunnii. The molecular weight, sugar content and selenium content of SeCPS-II were 4.12 × 103 kDa, 17.89 μg g-1 and 90.75%, respectively. The structure of SeCPS-II was characterized using FT-IR, NMR, GC and GC-MS studies. SeCPS-II was composed of pyranose, which contained α-l-rhamnose, α-d-mannose, α-d-glucose and β-d-galactose at a ratio of 4.33 : 12.62 : 27.50 : 18.99. SeCPS-II contained α-(1 → 4)-d-glucose, α-(1 → 3)-d-glucose, β-(1 → 6)-d-galactose, α-(1 → 6)-d-mannose, and α-(1 → 4)-l-rhamnose, and the main chain was composed of α-(1 → 4)-d-glucose. An MTT assay indicated that SeCPS-II could influence the cell viability of SKOV-3 cells, H1299 cells and HepG2 cells in a dose-dependent and time-dependent manner. Morphologic changes (AO/EB staining and DAPI staining) and an MMP assay (JC-1 staining) of SKOV-3 cells treated with SeCPS-II were performed to research the antitumor activity of SeCPS-II. SeCPS-II could significantly induce apoptosis in SKOV-3 cells with typical apoptotic characteristics. To study the mechanism, the expressions of caspase-3, caspase-9, PARP, p53, Bax and Bcl-2 in SKOV-3 were studied via western blotting. SeCPS-II can stimulate the apoptosis of SKOV-3 cells through the p53-Bax-caspase pathway. Animal experiments revealed that SeCPS-II inhibits tumors within an ovarian tumor model rat, modeled with SKOV-3 cells. All the results indicated that selenium-enriched C. gunnii can be used to develop new selenium-containing additives.

摘要

硒(Se)已被确认为一种必需元素。动物和人类吸收并保留更多的有机硒而不是无机硒。生物转化是将无机硒转化为有机硒的主要途径。蛹虫草(Cordyceps gunnii),作为一种中国食用和药用真菌,具有多种生物功能。蛹虫草与硒通过液体发酵结合,获得富硒蛹虫草。从富硒蛹虫草中提取了一种新型硒多糖(SeCPS-II)。SeCPS-II 的分子量、糖含量和硒含量分别为 4.12×103 kDa、17.89μg g-1和 90.75%。使用傅里叶变换红外光谱(FT-IR)、核磁共振(NMR)、气相色谱(GC)和气相色谱-质谱联用(GC-MS)研究了 SeCPS-II 的结构。SeCPS-II 由吡喃糖组成,其中含有α-l-岩藻糖、α-d-甘露糖、α-d-葡萄糖和β-d-半乳糖,比例为 4.33:12.62:27.50:18.99。SeCPS-II 含有α-(1→4)-d-葡萄糖、α-(1→3)-d-葡萄糖、β-(1→6)-d-半乳糖、α-(1→6)-d-甘露糖和α-(1→4)-l-岩藻糖,主链由α-(1→4)-d-葡萄糖组成。MTT 测定表明,SeCPS-II 可在剂量和时间依赖性方式下影响 SKOV-3 细胞、H1299 细胞和 HepG2 细胞的细胞活力。通过 AO/EB 染色和 DAPI 染色以及 MMP 测定(JC-1 染色)对 SeCPS-II 处理的 SKOV-3 细胞进行形态学变化研究,以研究 SeCPS-II 的抗肿瘤活性。SeCPS-II 可显著诱导 SKOV-3 细胞发生凋亡,具有典型的凋亡特征。为了研究其机制,通过蛋白质印迹法研究了 SKOV-3 细胞中 caspase-3、caspase-9、PARP、p53、Bax 和 Bcl-2 的表达。SeCPS-II 可以通过 p53-Bax-caspase 途径刺激 SKOV-3 细胞凋亡。动物实验表明,SeCPS-II 可抑制 SKOV-3 细胞荷瘤模型大鼠中的肿瘤。所有结果表明,富硒蛹虫草可用于开发新的含硒添加剂。

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