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两种褐藻中多糖的分离分析及其生物活性研究
侯宁宁
学位类型硕士
导师张全斌
2017-05-23
学位授予单位中国科学院海洋研究所
学位授予地点青岛
学位专业海洋生物学
关键词莫氏马尾藻 多肋藻 多糖 抗凝血 成纤维细胞生长因子
摘要莫氏马尾藻(Sargassum maclurei)和多肋藻(Costaria coatata)是分别生长于南海和西北太平洋沿岸的海洋褐藻,对其多糖的研究非常有限。本文以莫氏马尾藻及多肋藻为研究材料,通过热水或稀酸提取方法进行粗多糖提取,并利用DEAE-Sepharose Fast Flow柱层析对粗多糖进行分级纯化。通过理化指标测定并结合红外光谱、核磁共振波谱等分析方法,对多糖的理化性质进行研究。结果表明莫氏马尾藻多糖是含有岩藻糖、糖醛酸及多种单糖的结构较复杂的硫酸化多糖;而多肋藻多糖为含有岩藻糖、半乳糖等多种单糖的结构复杂并且适度乙酰化的硫酸化多糖,其硫酸化位置多位于半乳糖及岩藻糖的C2位。
体外抗氧化活性结果表明莫氏马尾藻多糖清除自由基能力较强,具有良好的抗氧化活性;其抗氧化能力与浓度呈现正相关,且水提多糖的抗氧化能力要强于酸提多糖。
体外抗凝血活性研究表明:莫氏马尾藻多糖及多肋藻多糖都具有良好的抗凝血活性,且与剂量正相关,多肋藻多糖的抗凝血活性显著优于莫氏马尾藻。多糖主要抑制内源性凝血途径,并且其抗凝血活性随着硫酸根含量的增加而增强,而降解后的低分子多糖抗凝血活性明显降低,说明抗凝血能力与分子量及硫酸根含量相关。
以BaF3细胞体系测定莫氏马尾藻多糖及多肋藻多糖对FGF/FGFR(成纤维细胞生长因子/成纤维细胞生长因子受体)信号通路激活的作用。莫氏马尾藻多糖尤其是未经分级纯化的粗多糖及降解后的水提粗多糖对该信号通路激活作用较好;而多肋藻多糖对FGF/FGFR信号通路激活作用较明显,尤其是未降解多糖,且其对不同FGF/FGFR信号通路(主要是FGF-1,2,7,8,9,10)的激活作用有所不同,说明多糖对FGF/FGFR信号通路的激活作用与硫酸根含量及分子量相关。
其他摘要Studies on polysaccharides from two kinds of brown algae, Sargassum maclurei and Costaria costata were rarely concerned. In this study, the polysaccharides were extracted from S. maclurei and C. costata by hot-water or dilute acid extraction, and  purified through anion exchange chromatography with DEAE Sepharose FF-column. The structure and chemical characteristic were elucidated based on FT-IR, NMR and normal chemical analysis. Results showed that the polysaccharide from S. maclurei was a kind of sulfated heteropolysaccharide composed of fucose, glucouronic acid and several other monosaccharides. While the polysaccharide from C. costata was a complex and highly acetylated polysaccharide that sulfated mainly at C2 position of galactose and fucose residues.
The antioxidant activity of polysaccharide from S. maclurei was examined in vitro. Results showed that the S. maclurei polysaccharide had great antioxidant activity with strong ability to remove free radicals. Meanwhile, the antioxidant capacity was positively related to the concentration of polysaccharides, and the polysaccharides extracted by hot water had better antioxidant capacity.
The anticoagulant activities of polysaccharides from S. maclurei and C. costata were determined in vitro by the APTT, PT and TT assays using the sheep plasma. Results showed that they all had great anticoagulant activity, and polysaccharides from C. costata were better. The polysacchrides prolonged the APTT and TT, suggested they had anticoagulation activities by inhibiting  the intrinsic pathways. Meanwhile, the results showed a positive correlation between clotting time and sulfate content. While the low molecular weight components had no distinct prolongation of APTT, PT and TT, indicating that the molecular weight might also play an important role in the anticoagulant activity of polysaccharides from brown algae.
The BaF3 lymphoid cell lines were used to determine the activity of polysaccharides that to activate the fibroblast growth factors/ fibroblast growth factors receptors (FGF/FGFR) signaling. The results showed that the crude polysaccharides from S. maclurei and its degradation had great activity to activate the signal. And the polysaccharides from C. costata especially the fractions without degrading activated all the tested FGF/FGFR pairs greatly, while the effects of different components with different sulfate content to the FGF-1, -2, -7, -8, -9 and -10 were slightly different. It indicated that the FGF/FGFR signal activating activity was correspond with the sulfate group and the molecular weight.
学科领域生物学
语种中文
文献类型学位论文
条目标识符http://ir.qdio.ac.cn/handle/337002/136565
专题实验海洋生物学重点实验室
作者单位中国科学院海洋研究所
推荐引用方式
GB/T 7714
侯宁宁. 两种褐藻中多糖的分离分析及其生物活性研究[D]. 青岛. 中国科学院海洋研究所,2017.
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