Knowledge Management System Of Institute of Oceanology, Chinese Academy of Sciences
|Place of Conferral||海洋研究所|
|Keyword||多肋藻 褐藻多糖硫酸酯 结构表征 肺纤维化 免疫调节|
1. 用热水浸提的方法从多肋藻中提取了多肋藻总糖CCP。CCP经过DEAE-Sepharose Fast Flow凝胶柱洗脱获得三种分级组分F3、F4及F5。化学组分分析表明CCP是一种以半乳糖、岩藻糖为主，硫酸基含量为18.54%，分子量为149846 Da的复杂多糖。F3是一种各种单糖含量比较平均的、硫酸基含量极低的杂多糖，重均分子量为57264 Da；F4是以岩藻糖为主、甘露糖次之的硫酸化岩藻聚糖，重均分子量为74005 Da；F5是以半乳糖和岩藻糖为主、其他单糖含量极低的硫酸化半乳岩藻聚糖，其重均分子量为75859 Da。
Costaria costata belongs to Costaria Greville, is a large annual brown algae with a long history of consumption, mainly in the northern Pacific coast, Hokkaido, Japan and the Great Peter Bay. For many years, most of the research on C. costata has been focused on ecological studies and artificial culture, but the study of its polysaccharides is still not comprehensive enough and needs further research. In this study, the extraction, graded purification and degradation of fucoidan from C. costata (CCP) were investigated. The chemical properties and structure of CCP were analysed, and the biological activity was also systematically investigated to lay the foundation for the comprehensive exploitation of C. costata.
In this study, the CCP was isolated from C. costata, and three main graded fractions of CCP were obtained by graded purification named F3, F4, F5. The chemical properties of CCP, F3, F4 and F5 were investigated, and the structure of F5 was analysed. The anti-pulmonary fibrosis activity, anti-tumour activity and immunomodulatory activity of CCP, F3, F4 and F5 were evaluated by in vitro and in vivo models, respectively. The main experimental results are as follows:
1. CCP was extracted by hot water extraction and further eluted on a DEAE-Sepharose Fast Flow gel column to obtain three graded fractions F3, F4 and F5. Chemical composition analysis showed that CCP is a complex polysaccharide with galactose and fucose as the main components, with a sulfate group content of 18.54% and a molecular weight of 149846 Da. F3 is a heterogeneous polysaccharide with an average content of various monosaccharides and a very low content of sulfate group, with a molecular weight of 57264 Da; F4 is a sulfated fucose with a molecular weight of 74005 Da, mainly fucose, followed by mannose; F5 is a sulfated polysaccharide with a molecular weight of 75859 Da, mainly galactose and fucose, with a very low content of other monosaccharides.
2. The structural analysis of F5 was carried out by NMR, IR spectroscopy and mass spectrometry. The results of the spectral analysis showed that there were mainly eight kinds of heterocapitated carbons in F5, and the polysaccharide configuration was α-type; F5 mainly contained three types of polysaccharide fragments, separately, they are sulfated-fucose oligosaccharide, sulfated-galactose oligosaccharide and sulfated fucose-galactose oligosaccharide; the oligosaccharide polymerization degree was 1~9, and it contained different numbers of sulfate group substitutions, and the sulfate group substitutions mainly occurred at the C-2 and C-3 positions, and there was no sulfate group substitution at the C-4 position.
3. The anti-fibrotic activity of CCP was investigated in vivo using a mouse model of pulmonary fibrosis, and CCP was found to have in vivo anti-fibrotic activity. CCP treatment was able to reduce inflammatory factor expression and inhibit collagen deposition in mice with pulmonary fibrosis. Further studies revealed that CCP could resist TGF-β1-induced epithelial-mesenchymal transition by reducing the expression of COL2A1 and p-Smad2/3 in vitro.
4. In the study of the anti-tumor activity and immunomodulatory activity of CCP and its graded fractions, it was found that F3, F4, F5 and CCP could inhibit the growth of lung cancer cells and pancreatic cancer cells and promote apoptosis in A549 and PANC-1 cells. In the evaluation of immunomodulatory activity, it was found that F4, F5 and CCP did not have pro-inflammatory activity at low concentrations, while at high concentrations F4, F5 and CCP all had some pro-inflammatory activity. F3 had anti-inflammatory activity at both low and high concentrations; the remaining fractions did not have anti-inflammatory activity.
From the above experiments, we conclude that Fucose Sulfate of D. multifida has good potential for anti-pulmonary fibrosis, anti-pulmonary tumour, anti-pancreatic tumour and immunomodulatory applications.
|MOST Discipline Catalogue||工学|
|Table of Contents|
|魏思洁. 多肋藻褐藻多糖硫酸酯的制备、结构表征及生物活性评价[D]. 海洋研究所. 中国科学院大学,2023.|
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