【作者】 曹荣；

【导师】 王加启； 雒秋江；

【作者基本信息】 新疆农业大学 ， 动物营养与饲料科学， 2007， 硕士

【摘要】 本论文通过在围产期奶牛日粮中添加维生素D₃,研究了维生素D₃对奶牛血常规、免疫球蛋白及CD4⁺、CD8⁺的影响,并通过体外培养围产期奶牛外周血淋巴细胞,探讨了不同浓度1,25-（OH）₂D₃对细胞增殖、细胞分泌IFN-γ以及免疫球蛋白的影响。试验一选用16头胎次接近的围产期奶牛,采用完全随机试验设计,研究日粮中额外添加维生素D₃（VD₃:10万IU/头.d）对围产期奶牛血常规、免疫球蛋白和T细胞亚群CD4⁺、CD8⁺的影响。结果表明,在日粮中额外添加10万IU/头.d VD₃可以极显著提高外周血中红细胞、白细胞、血红蛋白的量（P<0.01）。并且血小板数目、淋巴细胞百分比、单核细胞百分比、中性粒细胞绝对值和单核细胞绝对值均有不同程度提高（p>0.05）。添加VD₃组IgG、IgM和IgA均高于对照组（P>0.05）,T细胞亚群CD4+、CD8+的数量都极显著高于对照组（P<0.01）。本试验结果表明,日粮中额外添加10万IU/头.d VD₃可以缓解围产期奶牛免疫球蛋白和免疫细胞下降的现象,增强奶牛非特异性免疫功能。试验二采用完全随机试验设计,研究添加0、0.01、0.1、1、10和100nM 1,25-（OH）₂D₃对体外培养的围产期奶牛外周血淋巴细胞免疫功能的影响。结果表明,添加不同浓度的1,25-（OH）₂D₃对淋巴细胞增殖表现出一定程度的抑制作用（p>0.05）,显著抑制了ConA和PWM刺激的淋巴细胞IFN-γ的分泌。0.01～100nM 1,25-（OH）₂D₃剂量依赖性抑制了ConA和PWM诱导的淋巴细胞IFN-γ的产量。当1,25-（OH）₂D₃浓度达到0.1nM、1nM时与对照组相比分别显著（p<0.05）和极显著（p<0.01）抑制ConA诱导的淋巴细胞IFN-γ的产量。以PWM激活的细胞在培养48h、72h, 0.1～100nM1,25-（OH）₂D₃均极显著抑制IFN-γ的产生（p<0.01）。0.01nM 1,25-（OH）₂D₃对IFN-γ的抑制作用相对较小（p>0.05）,在培养48h后并未表现出显著的抑制作用,但培养72h后也极显著的抑制了IFN-γ的产生。低浓度的1,25-（OH）₂D₃（0.01、0.1nM）促进免疫球蛋白的产生,高浓度表现抑制作用。其中在培养的第7d, 0.01nM组IgM浓度显著高于对照组,培养第9d 0.01、0.1nM组IgA、0.01nM组IgM均极显著高于对照组。1,25-（OH）₂D₃的添加量在1～100nM范围内对淋巴细胞IgA和IgM的产生表现出抑制作用,且随添加量的增大抑制增强。0.1、0.01nM添加组在培养第3d以后表现出促进IgG产生的作用,但与对照组相比差异不显著,添加高浓度的1,25-（OH）₂D₃抑制了IgG的产生,10、100nM添加组在培养3d后,与对照组相比差异显著,1nM添加组在整个培养期间,IgG的产量与对照组相比无显著差异。更多还原

【Abstract】 Two trials were conducted to study the effect of vitamin D₃ on the immune function of periparturient dairy cattle. One is effect on Blood routine ,Immunoglobulins（Ig） , T helper （CD4⁺） and T-cytotoxic/suppressor （CD8⁺） of peripheral blood cow in vivo,and another is study that effects of diverse concentration 1,25-（OH）₂D₃on the proliferation, secretion of interferon-γ（IFN-γ） and Immunoglobulins of peripheral blood lymphocytes（PBLs） from periparturient dairy cattle in vitro.Trial 1:Sixty Holstein breed cows were randomly assigned to experiment or control peripartum diet to investigate the effect of Vitamin D₃ on Blood routine ,Immunoglobulins（Ig） , T helper （CD4⁺） and T-cytotoxic/suppressor （CD8⁺） of peripheral blood. The results showed that RBC、WBC、HGB were increased by supplement 105IU/d VD₃ in diet of every cow, and the difference is significant （P<0.01）. Compear to control, the PLT, LY%, MO%, number of GR, number of MO also were enhanced, but the difference is not significant （P>0.05） . At the same time, the IgG、IgM and IgA were higher than control , but the difference is not significant （P>0.05） . However, CD4+、CD8+T cells in experiment treatment were significantly higher than in control （P<0.01）. The results suggest that Ig and T cell subsets decreased in periparturient dairy cattle can be mitigated by supplement VD₃ in diets.Trial 2:This research investigated the in vitro effects of 0, 0.01, 0.1, 1, 10 and 100nM 1,25-（OH）₂D₃ on the proliferation, secretion of interferon-γ（IFN-γ） . The results suggest that 1,25-（OH）₂D₃ have no effect on proliferation of lymphocytes, but inhibited synthesis of IFN-γby ConcanavalinA and Pokeweed mitogen activated PBLs in a dose-dependent fashion. 0.1 nM and 1nM 1,25-（OH）₂D₃ both significantly suppressed （0.1 nM:p<0.05;1nM:p<0.01）synthesis of IFN-γby ConcanavalinA activated PBLs. And 0.1¹00nM 1,25-（OH）₂D₃ have significant suppressed effect on synthesis of IFN-γby Pokeweed mitogen activated PBLs in 48h and 72h（p<0.01）. However, 0.01nM 1,25-（OH）₂D₃ have no significant effect on IFN-γin 48h by ConcanavalinA and Pokeweed mitogen activated PBLs,but were significantly suppressed（p>0.05）in 72h.0.01,0.1nM 1,25-（OH）₂D₃ inhenced Ig production, but 1,10,100nM inhibited ones. IgM production in group of 0.01nM were significant higher than control in 7d （p<0.05）. And IgA production in group of 0.01,0.1nM ,IgM production in group 0.01nM also significant increased（p<0.01）in 9d. However, 1¹00nM 1,25-（OH）₂D₃ inhibited synthesis of IgA and IgM in a dose-dependent fashion. 0.01, 0.1nM 1,25-（OH）₂D₃ induced IgG production,but have no significant difference in 3d （p>0.05）. 10、100nM1,25-（OH）₂D₃ have significant suppressed IgG production 3 days later.However IgG production supplyment 1nM 1,25-（OH）₂D₃ has not significant difference with control group during the culture .更多还原