| Fucoidans are one of the important active components of Sargassum fusiforme, and have extensive biological activities. The research issues are the immunological activities and mechanism of fucoidans. But due to the complexity of its composition, similar polarity, and the distribution of its molecular weight is wide, the work of separating and purificating systematically the fucoidans and filtering the active components was very little, which made the molecular mechanism of immunoregulatory activity of fucoidans from S. Fusiforme is not clear, so far. In order to solve this problem, we need to do the work of separating and purificating the fucoidans from S. Fusiforme systematically and filtering the components, which have immunoregulatory activety.In this article,water-soluble fucoidans(SFPF) were isolated by using the methods of hot water extraction, alcohol precipitation and salting from S. Fusiforme. SFPF-1, SFPF-2 and SFPF-3 were purified from SFPF by DEAE Sepharase CL-6B and Sephacryl S400 column chromatography. The basic stucture information of these four compenents were analyzed by determinating the contents of total carbohydrate, uronic acid, protein and sulfate groups, and using the method of PMP-HPLC and HPGPC studied the monosa-ccharide composition and the average molecular weight.Furthermore, the immunoregulatory activity of these four components were also studied by detecting the influence of these components for the function of spleen lymphocytes and the peritoneal macrophages in mice. In vitro, different dose(10 μg/ml, 100 μg/ml, 1000 μg/ml) of these components cutured with spleen lymphocytes and macrophages witch were from normal mice and then detected their related activity. In vivo, SFPF and SFPF-2 at the doses of 100 mg/kg, 200 mg/kg and 400 mg/kg were given to mice by intragastric administration, successive medication for 7 days, and then the effects of SFPF and SFPF-2 were investigated in mice by detecting the function of spleen lymphocytes and macrophage.Results of the basic physical and chemical properties of SFPF, SFPF-1, SFPF-2 and SFPF-3 show that, the total carbohydrate content of these four components are 65.4%, 77.8%, 81.8% and 77.8% respectively; uronic acid content are 12.4%, 17.0%, 14.7% and 7.5% respectively; protein content are 1.4%, 1.6%, 0.4% and 0.3% respectively; sulfate content are 9.3%, 2.9%, 5.6% and 18.3% respectively; the molecular weight of SFPF-1, SFPF-2, SFPF-3 are 4.4 x104 Da, 2.4 x104 Da, 6.0 x104 Da. Monosaccharide composition is mainly composed of man, gal, xyl and fuc, containing a few Gal A, the proportion of this four main monosaccharide of SFPF is 2.1: 10.2: 5.6: 81.0, SFPF-1 is 1.3:4.6: 8.0: 82.5, SFPF-2 is 2.4: 13.3: 3.1: 80.6 and SFPF-3 is 1.0:10.0: 2.2: 84.1.Active results show that through the in vitro drug delivery, SFPF, SFPF-1,SFPF-2, SFPF-3 can promote both spleen T lymphocyte proliferation induced by Con A and B lymphocyte proliferation induced by LPS, also as an original split promote the mice spleen lymphocyte proliferation. SFPF and its purificated component also can enhance the killing activity of NK cells to Yac-1 and promote the phagocytosis of macrophages, enhance the cytotoxicity of macrophages to L929 cell, and a dose dependent.Through gastric with SFPF and SFPF-2, found that SFPF and SFPF-2 could significantly promote lymphocyte proliferation induced by Con A and LPS, and as an original split promoted lymphocyte proliferation. Meanwhile,SFPF and SFPF-2 have a significant impact on the killing activity of NK cell and the cytotoxic activity of macrophages cell, and enhance the phagocytic activity of macrophage phagocytic for neutral red. |
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