中国农业科学 ›› 2010, Vol. 43 ›› Issue (17): 3645-3652 .doi: 10.3864/j.issn.0578-1752.2010.17.019

• 兽医 • 上一篇    下一篇

泰山松花粉多糖对小鼠免疫增强作用的研究

魏凯,孙振红,谭燕玲,王慧,王新建,朱瑞良

  

  1. (山东农业大学动物科技学院)
  • 收稿日期:2010-04-06 修回日期:2010-06-23 出版日期:2010-09-01 发布日期:2010-09-01
  • 通讯作者: 朱瑞良

Research on the Effect of Immune Enhancement of Taishan Pine Pollen Polysaccharide in Murine

WEI Kai, SUN Zhen-hong, TAN Yan-ling, WANG Hui, WANG Xin-jian, ZHU Rui-liang
  

  1. (山东农业大学动物科技学院)
  • Received:2010-04-06 Revised:2010-06-23 Online:2010-09-01 Published:2010-09-01
  • Contact: ZHU Rui-liang

摘要:

【目的】研究泰山松花粉多糖对健康小鼠免疫功能的增强作用以及对免疫抑制小鼠的免疫恢复作用。【方法】用改进的水提醇沉法提取泰山松花粉粗多糖。取清洁级昆明小鼠(雌性)160只,随机分为8组,3个多糖组(Ⅰ,Ⅱ,Ⅲ),3个环磷酰胺(Cytoxan, CTX)-多糖组(Ⅳ,Ⅴ,Ⅵ),另外2组为CTX对照组(Ⅶ)和正常对照组(Ⅷ)。Ⅰ、Ⅳ组多糖注射剂量为400 mg?kg-1,Ⅱ、V组多糖注射剂量为200 mg?kg-1,Ⅲ、Ⅵ组多糖注射剂量为100 mg?kg-1,从第1天连续皮下注射7 d;Ⅳ、Ⅴ、Ⅵ、Ⅶ组于第1天每只每次腹腔注射75 mg?kg-1 CTX,隔1天注射一次,共计3次;Ⅷ组皮下注射生理盐水。第8天,所有小鼠腹腔注射1头羽份的ND冻干苗,分别在免疫后3、10和15 d采取血液、脾脏和小肠,用β微量法检测血清抗体、全自动血细胞分析仪测定外周血淋巴细胞比率、流式细胞术检测脾淋巴细胞转化率,ELISA法检测小肠IgA含量。【结果】Ⅰ、Ⅱ、Ⅲ组各指标均显著高于Ⅷ组(P<0.05),其中Ⅰ、Ⅱ组与Ⅷ组相比差异极显著(P<0.01)。Ⅳ、Ⅴ组极显著高于Ⅶ组(P<0.01),Ⅵ组显著高于Ⅶ组(P<0.05),Ⅳ组结果与Ⅷ组差异不显著。并且免疫后10d的各组显著高于免疫后3d的各对应组(P<0.05),与免疫后15d的差异不显著。【结论】泰山松花粉多糖能提高正常小鼠和免疫抑制小鼠的抗体水平、血液淋巴细胞比率、脾脏淋巴细胞转化率以及小肠IgA含量,200 mg?kg-1剂量的效果即非常显著,400 mg?kg-1剂量能使CTX诱导的免疫抑制完全恢复到正常水平。

关键词: 泰山松花粉多糖(TPPPS), 环磷酰胺, 抗体, 免疫增强, 淋巴细胞, IgA

Abstract:

【Objective】 An 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. 【Method】 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-1, polysaccharide dose of group Ⅱ and V was 200 mg?kg-1, polysaccharide dose of group Ⅲ and Ⅳ was 100 mg?kg-1, the mice were administered by intraperitoneal injection for 7 days from the 1st day. GroupⅤ, Ⅵ, Ⅶ were injected with CTX (cyclophosphamide) intraperitoneally at 75 mg?kg-1 on 1st day, once every other day, a total of three times. GroupⅧ 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 taken and the relevant indicators were inspected on the 3rd, 10th, and 15th day after immunization. The lever of systemic antibody was detected by ΒMicrotest 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】 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 Group Ⅶ 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 immunization.【Conclusion】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-1 dose TPPPS was obvious, 400 mg?kg-1 dose TPPPS could completely restore CTX-induced immune suppression to the normal level.

Key words: Taishan pine pollen polysaccharide (TPPPS), CTX, antibody, immune enhancement, lymphocyte, Ig A