| Macrophage is a kind of important immune cells that widely distributed in the body. Macrophages mainly originated from the bone marrow precursor cells. Macrophage can be activated by outside stimuli, and the phagocytosis of the activated macrophages enhanced greatly which can kill pathogens directly through phagocytosis or indirectly by the secretion of inflammatory cytokines(eg. TNF-a, IL-6, and IL-lβ), NO, and reactive oxygen species, which formed the first line of defense of the organsim. Macrophages can also through the engulf pathogens and present antigen, so as to induce the body’s immune response against the exogenous pathogens. Because of Seaweed polysaccharides has many kinds of biological activity, such as such as antitumor, antiviral, antioxidant, hypolipidemic, immune regulation, etc. it was widely studied in recent years. Seaweed was now a lot of artificial breeding, rich in resources, explore its biological active ingredient, development of functional food in order to achieve high value utilization is of great significance.In this study, porphyran and polysaccharides from Gracilaria lemaneiformis were obtained by water decoction and alcohol precipitation, and purified by DEAE-cellulose column chromatography orderly. The carbohydrate, SO42-, uronic acid of porphyran are 81.20%, 14.67%, 7.35%. The carbohydrate, SO42-, uronic acid of polysaccharides from Gracilaria lemaneiformis are 81.19%, 9.33%, 8.20%. RAW264.7 macrophage was used to study the biological activity of porphyran and polysaccharides from Gracilaria lemaneiformis.. MTT test showed that both polysaccharide can effectively promote the proliferation of RAW264.7, enhance cell viability.In a certain concentration scale(5~100 μg/m L) for 12 hours, the proliferation of RAW264.7 caused by porphyran was dose-dependent. The cell viability reached the highest when incubated with porphyran of 40 μg/m L for 24 hours. The concentrations within the scope of 12.5 to 200 μg/m L of polysaccharides from Gracilaria lemaneiformis can significantly enhance the viability of RAW264.7 cells, and there was no significant difference between different concentrations. Neutral red uptake experiments show that porphyran and polysaccharides from Gracilaria lemaneiformis could significantly enhance the phagocytic ability of the macrophages. The phagocytic ability of treatment groups, 25 μg/m L, 50 μg/m L and 100 μg/m L of porphyran, were respectively 1.8, 2.1, 2.5 times in the control group, and the results of polysaccharides from Gracilaria lemaneiformis were 1.7, 2.0, 2.4 times. In addition, porphyran and polysaccharides from Gracilaria lemaneiformis could effectively promote macrophage RAW264.7 to release NO, TNF-α, IL-6. The serection of NO were 34.76 μmol/L, 49.20 μmol/L and 53.18 μmol/L respectively after treated with different concentrations of porphyran(25 μg/m L, 50 μg/m L and 100 μg/m L), it was remakablely increased compared to control group. And the same effects on TNF-α( 136.76 ng/L, 147.08 ng/L, 141.72 ng/L) and IL-6( 147.17 ng/L, 160.55 ng/L, 155.13 ng/L) were revealed. After incubating with different concentrations(50 μg/m L, 100 μg/m L, 200 μg/m L) of polysaccharides from Gracilaria lemaneiformis, the serection of NO were 34.76 μmol/L, 49.20 μmol/L, 53.18 μmol/L, and the serection of TNF-α were 136.76 ng/L, 147.08 ng/L and 141.72 ng/L respectively, and the serection of IL-6 were 147.17 ng/L, 160.55 ng/L and 155.13 ng/L respectively, and the results was remakablely increased compared to control group. The results showed that porphyran and polysaccharides from Gracilaria lemaneiformis promoted RAW264.7 to release NO mainly through JNK and JAK2 signaling pathway.In summary, porphyran and polysaccharides from Gracilaria lemaneiformis can effectively activate the macrophage RAW264.7, promote the cell proliferation, enhance their consuming ability, regulate secretion of cytokines released by RAW264.7. |
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