| Summary: | 碩士 === 國立海洋大學 === 食品科學系 === 89 === Marine algae contain large amounts of characteristic polysaccharides such as alginate, funcoidan, carrageenan, and agarose have physiological activity. It have reported that the enzyme-degraded polysaccharide fraction from Gracilaria verrucosa (GWS), which is a material for the agar industry , increased the number of PEC, phagocytic activity and oxygen radical activity ofmice in vivo (Yoshizawa et al., 1996). In our studies, screening of biological activity substances were performed by two treatments.Gracilaria tenuistipitata was extracted with Phosphate buffer and 70% amonium sulfate precipitation (GWS-A) and 0.1N HCl, thenboiled with hot-water (100 ℃, 30 min) (GWS-H). The cell proliferation activation was used as indicators of function. PBS extracts of Gracilaria tenuistipitata promoted cell proliferation, Immunoglobulin production and cell aggregation toward human hybridoma cell lines HB4C5, and promoted cell proliferation towardmacrophage-like cell lines (UM and J774.1) and PEC, spleen cell of C57BL/6J mice (♀, 9 weeks).In agglutination assay, Sheep red blood cells were agglutinated by GWS-A, and the agglutinating activity of GWS-A was inhibited by heating at 121oC for 15 min, trypsin and saccharide. Cell proliferation activity remained about 70%-80%, when being treated at 100 oC for 60min or 121 ℃15min. By using DEAE anion-exchange chromatography, the isolatedbioactive factor was found the protein fraction at 30% and 50%, shown strong both growth promoting and immunostimulating effect toward HB4C5, but their molecular weight and structure would be studed further. 0.1N HCl extracts (GWS-H) inhibited the growth of chronic myelogenous leukemia (K562), acute promyelocytic leukemia (HL-60), human hybridoma (HB4C5), breast cancer (MCF-7) and colon cancer (HT-29), but not sensitivetoward B16. Acid extracts (GWS-H) cannot prevent the growth ofPEC, spleen cell of C57BL/6J mice (♀, 9 weeks),and the extractsare considered to be non-toxic. In vitro, we proved the fraction ofoligosaccharide obtained from enzyme from small abalone treated GWS-H inhibited HL-60 and HB4C5 is better than GWS-H. For in vivo assay, mice (C57BL/6J) were i.p. administered GWS-H (4 mg/mouse, 8 mg/mouse) and PEC were harvested 2 to 5 days later. GWS-H increased the number of PEC, NK cell activity and phagocytosis of granulocyte from peripheral blood of mice at 3 to 4 days later. BALB/c mice were fed with different dose of GWS-H. The results showed GWS-H did not affect the growth curve and food efficiency by non-specific and specific immune response. GWS-H significantly increased serum antibody IgG and decreased liver lipid peroxidation. Furthermore, with ConA, PHA and LPS enhance promote spleen cell proliferation. In antitumor, GWS-H markedly inhibited growth of B16 and increased prolongation ratio. These result obtained in this study indicated that the extract of Gracilaria tenuistipitata should be good candidate for health food development.
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