| Polysaccharide abundant in Anoectochilus formosanus is one important active ingredient. It has a wide range of pharmacological activity. At present, the research on the extraction methods and immunologic enhancement of Anoectochilus formosanus Polysaccharide has made some progress, but the relevant mechanism research is still rarely reported. In this study, the genuine regional medicinal materials of Fujian Anoectochilus formosanus was taken as the material. It was made into ultra-fine powder and then selected by 300 mesh sieve. With water extracting-alcohol precipitating method combined with ultrasonic , the Anoectochilus formosanus polysaccharide from superfine powder could be extracted. Through the single factor test, the optimum extracting conditions for polysaccharide could be determined, at the same time, polysaccharides could be purified preliminarily. Phenol-sulfuric acid method could be taken to determine the content of polysaccharides. The immune adjustment effect on the immunosuppression mice caused by cyclophosphamide and the healthy one, which were administrated by different doses of Anoectochilus formosanus polysaccharide , could be inspected by the determination of mice immune organs and immune cells, immune molecules indicators.Test results are as follows:1. The single factor experiment results showed that the optimum extraction conditions for extracting Anoectochilus formosanus polysaccharide were ultrasonic power 80KHz,70℃,ratio of sample to solution 1:30,1h,twice . The average yield of Anoectochilus formosanus polysaccharide was 7.14%. With Phenol - sulfuric acid method, the total Anoectochilus formosanus polysaccharide content amounted to 87.62%.2. The immune regulation of Anoectochilus formosanus polysaccharides on the immunosuppressed mice2.1The effect of Anoectochilus formosanus polysaccharide on immune organs on immunosuppressed miceAnoectochilus formosanus polysaccharides could improve the weight of immunosuppressive mice. Spleen indexes and thymus indexes of three doses were significantly higher than the CY model group (P<0.05), in which the spleen weight of the low dose group and high dose group were significantly higher than CY model group (P<0.05), and thymus weight of high-dose group was significantly higher than the CY model group (P<0.05). That indicated polysaccharide could promote growth of immune organ in immunosuppressive mice2.2 The effect of Anoectochilus formosanus polysaccharide on immune cells of immunosuppressed miceCarbon clearance test results ,which showed that the phagocytic indexes of small dose and median dose group were significantly higher than the CY model group (P <0.05), indicated that both the two dose levels could promote the phagocytosis of immunosuppressive mice; mice Spleen lymphocyte proliferation test results by MTT method showed that the optical density value of three dosage groups were significantly higher than the CY model group (P<0.05), thus it suggested that Anoectochilus formosanus polysaccharides could stimulate lymphocyte the proliferation of immunosuppressive mice.2.3 The effect of Anoectochilus formosanus polysaccharide on the immune molecules of immunosuppressed miceThe level of cytokine detected by ELISA kit suggested that the TNF-α, IFN-γof three groups were higher than CY model group, and the difference was significant (P <0.01), the IL-6 levels of medium dose group and high dose group were significantly higher than CY model group (P<0.05). This result suggested that the medium and high dose groups could promote the secretion of cytokines of immune suppressed mice and enhance its Secretion too. Turbidimetric method was applied to determine the levels of immunoglobulin IgA, IgM, IgG with the C3, C4 content ,then the result indicated that IgA of the high-dose group was significantly higher than that of CY model group(P<0.05); the IgM level of medium and high dose groups were significantly higher than the CY model group (P<0.05); the medium dose IgG group was significantly higher than CY model group (P<0.05). besides, the C3 levels of the medium and high dose groups were significantly higher than that of CY model group (P<0.05); in addition, the C4 contents of medium and low dose groups were significantly higher than that of CY model group (P<0.05).These Descriptions reflected that Anoectochilus formosanus polysaccharide could promote the secretion of immune factors in the immunosuppressed mice.3.The immunomodulatory effects of Anoectochilus formosanus polysaccharide on the health mice3.1 The effect of Anoectochilus formosanus polysaccharide on immune organs of healthy miceAll of the three dose groups of Anoectochilus formosanus polysaccharide could improve the weight of healthy mice, spleen weight of three dose groups were significantly higher than the control group (P<0.05),and the thymus weight of low and high dose groups were significantly higher than the control group (P<0.05); the spleen index of high-dose group was significantly higher than the control group (P<0.05); the thymus index of medium and high dose groups were significantly higher than the control group(P<0.05).Such results indicated Anoectochilus formosanus polysaccharide could improve the weight of immune organs and immune organ indexes of healthy mice.3.2 The effect of Anoectochilus formosanus polysaccharide on immune cells of healthy miceCarbon clearance test results ,which showed that the high and medium dose groups phagocytic indexes were significantly higher than that of control group (P<0.05), demonstrated that the two dose groups could enhance macrophage phagocytosis of health ones; the results of lymphocyte proliferation test by MTT method, which showed there was no significant difference in OD values amongst three dose groups and the control group, revealed that the impact of Anoectochilus formosanus polysaccharide on lymphocyte proliferation was not significant.3.3 The effect of Anoectochilus formosanus polysaccharide on immune molecules of healthy miceResults of serum cytokines in healthy mice with ELISA kit manifested that, polysaccharide of the three dose groups played no significant role in the content of IL-6,and the levels of TNF-α, IFN-γin the medium and high dose groups were significantly higher than the control group (P<0.05).The levels of serum IgA, IgM, IgG and C3, C4,determined by turbidimetry in healthy mice, showed that IgA, IgM content of the three dose groups showed no significant difference compared with control group;in the high dose group,IgG was significantly higher than the control group (P<0.05); C3 of high-dose group was significantly higher than the control group too (P<0.01); C4 of medium dose group was significantly higher than the control group (P<0.05).The results above showed that Anoectochilus formosanus polysaccharide could promote the secretion of immune factors in healthy mice.
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