陈柳君,宁亚静,康彩峰,等.羊栖菜多糖的提取纯化及免疫活性初步探究[J].中国海洋药物,2017,36(3):81-88. |
羊栖菜多糖的提取纯化及免疫活性初步探究 |
Study on Extraction, Purification and Immunobiological Activity of Sargassum fusiforme Polysaccharides |
投稿时间:2016-10-15 修订日期:2017-03-03 |
DOI: |
中文关键词: 羊栖菜 多糖 纯化 结构分析 免疫活性 |
English Keywords:Sargassum fusiforme polysaccharide purification structure analysis immune activity |
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中文摘要: |
目的 提取纯化羊栖菜多糖,初步研究其理化性质及免疫活性。方法 热水煮提、乙醇沉淀多糖、CaCl2法除去褐藻胶、DEAE-纤维素脱色,经DEAE-Sephaose CL-6B和SephacrylTM S-400 HR分级纯化,得到3个多糖级分SFPF-1、SFPF-2和SFPF-3。高效液相色谱法(HPLC)、紫外分光光度计(UV)、红外光谱(IR)对多糖的结构进行初步分析;体内测定多糖对小鼠脾脏和胸腺指数的影响,体外检测多糖对淋巴细胞活性及对巨噬细胞吞噬能力等影响初步研究其免疫活性。结果 3个多糖级分的分子量在6.01×104~7.7×104 Da,三者均主要由岩藻糖硫酸酯组成,并且SFPF-2的硫酸根含量高达32.8%,而蛋白质和糖醛酸含量较低;体内实验表明多糖显著影响小鼠脾指数和胸腺指数;体外实验表明3个多糖对Raw264.7细胞的增殖能力和吞噬能力影响不一,但都明显促进TNF-α的分泌。结论 羊栖菜多糖SFPF-1、SFPF-2和SFPF-3均含有较高的硫酸根,并且在小鼠和细胞实验中具有显著的免疫刺激功能。羊栖菜多糖可以作为一种免疫刺激剂,具有较好的临床应用价值和应用前景。 |
English Summary: |
Objective To investigate the physical and chemical properties and immunostimulatory activity of Sargassum fusiforme polysaccharides. Methods S. fusiforme polysaccharides were purified by a combined method involving hot-water extraction, treatment with CaCl2 and DEAE-Cellulose, DEAE-Sephaose CL-6B and SephacrylTM S-400 HR column chromatography. Three main fractions of purified polysaccharides, SFPF-1, SFPF-2 and SFPF-3, were obtained and the basic structural characteristics and some physical and chemical properties of these polysaccharides were analyzed by HPLC, UV and IR spectroscopy. The effects of the purified polysaccharides on the spleen and thymus indexes of mice and on the cytoactive and phagocytic activities of Raw264.7 cells were determined. Results SFPF-1, SFPF-2 and SFPF-3 had similar molecular weights. They were mainly composed of fucose sulfate (>80%). SFPF-2 had the highest sulfate content (32.8%), but only a small amount of protein and sugar acid. Mice administered with SFPF-2 clearly showed an increase in the weights of the thymus and spleen. The three polysaccharides exerted different effects on the proliferation and phagocytosis of Raw264.7 cells, but they all promoted the secretion of TNF-α from the cells. Conclusion S. fusiforme polysaccharides contained a high level of sulfate fucose, and had significant immune stimulate function in some extent when administered to mice or applied to cultured immune cells. They might act as an immunostimulant with great application prospects and clinical values. |
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