鸡脾转移因子对鸡肠黏膜免疫屏障与抗氧化功能的影响

发布时间：2019-06-22 10:22

【摘要】：禽类产品是我国人民肉类消费的重要来源。近30年来,我国居民对禽产品的消费需求不断增加。但是受国内养殖业技术水平的限制,家禽养殖常常遭受疾病、营养失衡、应激等影响,使得禽产品供应经常发生较大波动。因此,如何提高家禽健康水平,提升家禽生产成绩,保障禽产品稳定供应以满足居民需求是我国农业产业的重要课题。转移因子(Transfer factor, TF)是一组由T淋巴细胞释放的没有抗原性的小分子多肽、核苷酸、氨基酸类物质组成的混合液,近年来作为增强免疫制剂在家禽生产领域的应用引起了越来越多人的关注。本试验对出壳后5日龄雏鸡灌服0.1mg、0.25mg及1.0mg剂量的鸡脾转移因子,对照组灌服生理盐水,至40日龄时取十二指肠、空肠、回肠及直肠组织,从肠道黏膜组织学结构、细胞因子、MUC2(黏蛋白基因2)及TLR2/4 (Toll样受体2/4)表达及肠道抗氧化功能等途径研究了鸡脾转移因子的作用机制。试验同时研究了鸡脾转移因子在生产中的应用,分析了其对鸡群增重、免疫器官发育和新城疫治疗效果的影响。结果如下：1.鸡脾转移因子对鸡肠黏膜组织学结构影响1.0mg剂量鸡脾转移因子可以显著增加40日龄试验鸡小肠的绒毛高度、肠绒毛高度／隐窝深度比值(V/C)(P0.05)和杯状细胞密度(P0.05)；但各剂量鸡脾转移因子对直肠杯状细胞数目的影响与对照组相比较差异不显著。结果表明鸡脾转移因子可改善鸡小肠的黏膜屏障功能。2.鸡脾转移因子对鸡肠道细胞因子的影响1.0mg剂量的鸡脾转移因子显著提高40日龄鸡十二指肠、空肠、回肠和直肠的IL-10含量(P0.05)。0.25mmg剂量鸡脾转移因子显著提高试验组空肠、回肠、直肠IL-13含量(P0.05)。O.1mg和0.25mg剂量的鸡脾转移因子显著降低40日龄鸡十二指肠的IL-1水平(P0.05)。0.25mg和1.0mg剂量水平的鸡脾转移因子显著降低试验组十二指肠内TNF-α表达水平(P0.05)。结果表明鸡脾转移因子可提高鸡肠道抗炎因子表达和抑制促炎因子分泌,改善肠道黏膜免疫屏障功能。3.鸡脾转移因子对鸡肠道MUC2、TLR2及TLR4 mRNA表达的影响灌服1.0mmg剂量的鸡脾转移因子可显著增加40日龄试验鸡十二指肠、回肠、直肠MUC2 mRNA水平(P0.05)。灌服0.25mg和1.0mg剂量鸡脾转移因子均可以显著降低TLR2、TLR4基因在十二指肠(P0.05)、空肠(P0.05)、回肠(P0.01；P0.05)和直肠(P0.01；P0.05)的表达水平,且这种作用在一定程度上表现出剂量依赖性。4.鸡脾转移因子对鸡肠道抗氧化功能的影响灌服0.25mg鸡脾转移因子显著提高40日龄鸡十二指肠、空肠、直肠CAT活性(P0.05)。0.25mg剂量组的十二指肠、空肠GSH-Px活力与对照组相比显著升高(P0.05)。0.25mg和1.0mg剂量的鸡脾转移因子可显著降低十二指肠、空肠及回肠内MDA含量(P0.05)。0.25mg剂量的鸡脾转移因子可显著性提高十二指肠、回肠、直肠的T-AOC(P0.05)。结果表明鸡脾转移因子可提高鸡肠道组织的抗氧化功能。5.鸡脾转移因子对免疫器官发育及疾病抵抗能力的影响对5日龄雏鸡隔周灌服1.0mg剂量的鸡脾转移因子,40日龄试验组鸡体重显著高于对照组(P0.05)。雏鸡5日龄起每日灌服0.25mg剂量的鸡脾转移因子可以显著提高14日龄和22日龄脾脏与胸腺重量,极显著提高新城疫免疫后的抗体效价(P0.001)。6.鸡脾转移因子在养鸡生产中的应用大群试验结果显示鸡脾转移因子可有效提高规模化生产鸡群的成活率、个体体重,降低料肉比,提高养殖户的综合效益。对感染新城疫的生产鸡群应急处理显示,鸡脾转移因子可以显著提高鸡群的抵抗力,降低感染新城疫鸡群的死亡率,大大降低因疫病传播带来的损失。结论：鸡脾转移因子可以显著提高鸡小肠绒毛高度,增加鸡小肠V/C比值,诱导肠道干细胞增殖分化发育,增加杯状细胞密度,改善肠道黏膜机械屏障功能；改变肠道细胞因子的动态分布,促进肠道抗炎因子IL-10及IL-13的表达,降低促炎因子IL-1和TNF-α水平,改善肠道黏膜免疫屏障功能,进而提高鸡群抗病能力和生长性能。其作用机制可能与鸡脾转移因子提高肠道MUC2基因的表达水平、下调各个肠段TLR2及TLR4 mRNA水平和提高肠道抗氧化功能有关。
[Abstract]:Poultry products are an important source of meat consumption in our country. In the past 30 years, the demand for consumption of poultry products has increased in our country. However, by the limitation of the technical level of domestic industry, poultry breeding often suffers from diseases, nutrition imbalance, stress and so on, so that the supply of poultry products often fluctuates greatly. Therefore, how to improve the health level of the poultry, to improve the performance of the poultry production, to guarantee the stable supply of the poultry products to meet the needs of the residents is an important subject of the agricultural industry in China. Transfer factor (TF) is a group of mixed liquor consisting of small, non-antigenic, small-molecular, nuclear, and amino-acid-like substances released by T-lymphocytes, which has attracted more and more attention in recent years as an increase in the application of immune preparation in the field of poultry production. In the experiment, the chicken spleen transfer factor of 0.1 mg, 0.25 mg and 1.0 mg was given to the five-day-old chicks after the shell, and the control group was given physiological saline, and the duodenum, jejunum, ileum and rectum were taken from the intestinal mucosa, and the histological structure and the cytokines of the intestinal mucosa were taken from the intestinal mucosa. MUC2 (mucin gene 2) and TLR2/4 (Toll-like receptor 2/4) expression and intestinal anti-oxidation function were used to study the mechanism of chicken spleen transfer factor. The application of chicken spleen transfer factor in production was also studied, and its effects on the weight gain, immune organ development and the treatment effect of Newcastle disease were analyzed. The results are as follows:1. The effect of the chicken's spleen transfer factor on the histological structure of the chicken's intestinal mucosa was 1.0 mg, and the spleen transfer factor of the chicken was significantly increased by 40-day-old, the ratio of the depth of the intestinal villus to the depth of the crypt (V/ C) (P0.05) and the density of the cup-like cells (P0.05). However, the effect of each dose of chicken spleen transfer factor on the number of the rectal goblet cells was not significantly different from that in the control group. The results show that the chicken spleen transfer factor can improve the mucosal barrier function of the small intestine of the chicken. The effect of the chicken's spleen transfer factor on the intestinal cytokines of the chicken was higher than that of the chicken's spleen transfer factor of 1.0 mg, and the content of IL-10 in the duodenum, jejunum, ileum and rectum of the 40-day-old chicken was significantly increased (P0.05). The spleen transfer factor of the 0.25 mmg dose of chicken was significantly increased in the jejunum and ileum of the experimental group. The level of IL-1 in the duodenum of the 40-day-old chicken was significantly lower than that of the chicken's spleen transfer factor (P <0.05). The level of TNF-1 in the duodenum of the test group was significantly reduced by the chicken spleen transfer factor of 0.25 mg and 1.0 mg (P0.05). The results show that the chicken spleen transfer factor can improve the anti-inflammatory factor expression of the chicken intestinal tract and inhibit the secretion of the pro-inflammatory factor and improve the immune barrier function of the intestinal mucosa. The effect of chicken spleen transfer factor on the expression of MUC2, TLR2 and TLR4 mRNA in chicken's intestinal tract significantly increased the level of MUC2 mRNA in the duodenum, ileum and rectum of the 40-day-old test (P0.05). The expression level of TLR2 and TLR4 gene in the duodenum (P0.05), jejunum (P0.05), ileum (P0.01), and rectum (P0.01) and rectum (P0.01) and rectum (P0.01) was significantly reduced. The effect of the chicken's spleen transfer factor on the anti-oxidation of the chicken's intestinal tract was significantly increased in the duodenum, jejunum and rectum of the 40-day-old chicken (P0.05). The duodenum of the 0.25 mg dose group, The activity of GSH-Px in the jejunum was significantly higher than that in the control group (P0.05). The dose of 0.25 mg and 1.0 mg of the chicken spleen transfer factor could significantly decrease the content of MDA in the duodenum, jejunum and ileum (P0.05). The dose of 0.25 mg of the chicken spleen transfer factor could significantly increase the T-AOC of the duodenum, the ileum and the rectum (P0.05). The results show that the chicken's spleen transfer factor can improve the anti-oxidation function of the chicken's intestinal tissue. The effect of the chicken's spleen transfer factor on the development of immune organs and the resistance of the disease to the disease resistance was significantly higher than that of the control group (P0.05). The spleen and thymus weight of the spleen and thymus of the 14-day-old and 22-day-old chickens were significantly improved, and the antibody titer after the immunization of the newcastle disease was significantly improved (P 0.001). The results show that the chicken spleen transfer factor can effectively improve the survival rate of the chicken group, the individual weight, the ratio of the feed and the meat, and improve the comprehensive benefit of the farmers. The chicken-spleen transfer factor can obviously improve the resistance of the chicken group, reduce the mortality of the chicken group infected with the newcastle disease, and greatly reduce the loss caused by the transmission of the disease, according to the emergency treatment of the chicken group infected with the newcastle disease. Conclusion: The chicken spleen transfer factor can significantly improve the height of the small intestine of the chicken, increase the V/ C ratio of the small intestine of the chicken, induce the proliferation and differentiation of the intestinal stem cells, increase the density of the goblet cells, improve the mechanical barrier function of the intestinal mucosa, and change the dynamic distribution of the intestinal cytokines, The expression of IL-10 and IL-13 in the intestinal tract is promoted, the IL-1 and TNF-1 levels of the proinflammatory factors are reduced, the immune barrier function of the intestinal mucosa is improved, and the disease resistance and the growth performance of the chicken group are further improved. The mechanism of action may be related to the improvement of the expression level of the intestinal MUC2 gene, the down-regulation of the levels of TLR2 and TLR4 mRNA in the intestinal segments and the improvement of the anti-oxidation function of the intestinal tract.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S831

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