Research On The Extraction And Immune Enhancement Of Taishan Pine Pollen Polysaccharide

Pine pollen, one variety of pollen combining both edible and medicinal function in Chinese medical treasure-house, is regarded as the natural mini-nutrition storeroom. The unique efficacy of pine pollen is on record in ancient literatures of various Chinese dynasties. Pine pollen polysaccharide is composed of monosaccharides such as mannose, glucuronic acid, rhamnose, glucose, galactose, arabinose, and fucose etc.. Modern research has revealed that polysaccharide is one of the nutrient components that retain special-effects of pine pollen, with exceedingly high biological activity. There is a notable rise in the polysaccharide extraction quantity if wall-broken pine pollen is adopted. Polysaccharide was extracted from fresh Pinus massoniana pollen (sieved and break-walled) collected from Taishan region. Pepsin and Sevage reagent were used to remove protein in the extraction process, which ensured the high purity of polysaccharide.1 Research on the effect of immune enhancement of Taishan pine pollen polysaccharide in murineAn experiment was conducted to study the effects of immune enhancement caused by the Taishan pine pollen polysaccharides on healthy mice and the effects of immune recovery on immunosuppressed mice. The modified method of water extraction and alcohol precipitation of Ma Rongchi etc. was used to extract high-dose Taishan pine pollen polysaccharide (TPPPS). One hundred and sixty clean-level Kunming mice (female) were randomly divided into 8 groups: 3 polysaccharide groups (I, II, III), 3 Cytoxan (CTX)-polysaccharide groups (IV, V, VI), the CTX group (VII) and the control group (VIII). Polysaccharide doses of groupIand IV was 400 mg/kg, polysaccharide dose of group II and V was 200 mg/kg, polysaccharide dose of group III and IV was 100 mg/kg, the mice were administered by intraperitoneal injection for 7 days from the 1st day. Group V, VI, VII were injected with CTX intraperitoneally at 75 mg/kg on 1st day, once every other day, a total of three times. Group VIII was injected with physiological saline. On the 8th day, all mice were injected intraperitoneally a plume of ND vaccine. Samples of blood, spleen, small intestine were collected and the relevant indicators were inspected on the 3rd, 10th, and 15th day after immunization. The lever of systemic antibody was detected byβ-Microtest HA and HI, lymphocyte ratio was detected by automated hematology analyzer, spleen lymphocyte transformation rate was detected by flow cytometry, intestinal IgA levels were detected by ELISA method. Results showed the result of Group I, II, III were significantly higher than Group VIII (P<0.05). The differences between Group I, II and Group VIII was very significant (P<0.01). The results of group IV, V were higher than GroupVII very significantly (P<0.01), Group VI was significantly higher than Group VIII (P<0.05). The difference between Group IV and VIII was not significant. And the test results of the groups on the 10th day after immunity were significantly higher than that on the 3rd day (P<0.05) and not significant compared with the the groups on the 15th day after immunitzation.Concl sion: The extraction rate of Taishan pine pollen polysaccharide was 4.34% , the content of polysaccharide was 72.2% by using the method of water extraction and alcohol precipitation in this expermient. Different doses injection of Taishan pine pollen polysaccharide could improve normal and immunosuppressed mice's systemic antibody level, blood lymphocyte ratio, splenic lymphocyte proliferation, intestinal IgA level, the effect of 200 mg/kg dose TPPPS was obvious, 400 mg/kg dose TPPPS could completely restore CTX-induced immune suppression to the normal level.2 Effects of Taishan Pinus massoniana pollen polysaccharide on immune response of rabbit haemorrhagic disease tissue inactivated vaccine and on production performance of Rex rabbitsVaried doses of Taishan pinus pollen polysaccharide (TPPPS) and astragalus polysaccharide (APS) extracted by hot water extraction and ethanol precipiration method were added to the vaccine in order to prepare rabbit haemorrhagic disease (RHD) polysaccharide inactivated vaccine. 72 healthy Rex rabbits (rabbit haemorrhagic disease maternal antibody titer 0) each weighing 1 kg or so were selected and randomly allocated into 6 groups: 3 TPPPS vaccine groups ( I, II, III ), 1 APS vaccine control group (IV). And the other 2 groups were the vaccine-only control group (V) and the blank control group (VI). Each rabbit was injected with 1ml of the vacine liquids. TPPPS contents of the vaccine liquids in group I, II, and III were respectively 400 mg/ml, 200 mg/ml, 100 mg/ml, and APS content in group IV 400 mg/ml. Group V contained no polysaccharide in its liquid,and group VI was the equivalent physiological saline. The blood samples were then taken from the rabbits 0, 3, 7, 14, 21, 28, 35, 42 days after immunization. Serum antibody titer was determined byβ-microscopic method HA and HI, blood lymphocyte ratio by full automatic hematology analyzer, and peripheral blood lymphocyte proliferation rate by flow cytometry; at the same time average daily intake and average daily gain every week of the immuned Rex rabbits were measured to assess effects TPPPS exerted on their production performance. The results showed that each index in group I, II, III, and IV was higher than that in group V, especially group I, II, and IV, the distinction between which and group V was much more conspicuous (P<0.05); each index in group I was extremely statistically higher than that in group V (P<0.01); each index in group I was significantly higher than that in group II, III (P<0.05), and generally no significant differences were observed between group II and III. The overall level in group IV was slightly lower than that in group I. Each index in the polysaccharide groups reached its peak value later than the non-polysaccharide groups.The results suggested that any dose of TPPPS immune potentiator can enhance immunologic function and production performance of rabbits, and the amount of 400 mg has the most obvious efficacy. Furthermore, this immune potentiator can extend the immune peak period of rabbit haemorrhagic disease virus tissue inactivated vaccine and the growing peak period of Rex rabbits. TPPPS has generally higher efficiency than APS.