谷氨酰胺对断奶仔猪免疫应激调节作用及其分子机制的研究

发布时间：2018-05-06 14:26

本文选题：谷氨酰胺 + 断奶仔猪　；参考：《沈阳农业大学》2017年硕士论文

【摘要】：本论文研究日粮中添加谷氨酰胺对断奶仔猪生长性能、免疫应激调节作用及其分子机制。试验选取64头断奶仔猪,随机分为4组,每组4个重复,每个重复4头猪。采用2×2析因试验设计,日粮类型(基础日粮或1.2%谷氨酰胺日粮)和应激类型(注射LPS或生理盐水)。试验期28天。Ⅰ、Ⅱ组喂食基础日粮,分别在试验第14天和21天腹腔注射等量的生理盐水和LPS配置液(100μg/kgBW);Ⅲ、Ⅳ组在基础日粮中添加1.2%谷氨酰胺,分别在第14天和21天腹腔注射等量生理盐水和LPS配置液(100μg/kg BW)。每个处理组当天注射后3h后进行前腔静脉采血,分离血清,测定血清中免疫球蛋白及各种细胞因子的含量变化。在试验的第29天屠宰,取仔猪胸腺、脾脏及肠组织黏膜(十二指肠、空肠、回肠),采用荧光定量PCR技术检测组织中TLR4、HSP70、MMP-2、MMP-9基因在不同处理组中的表达差异。设Ⅰ组为本次试验的对照组,其研究结果显示:1、LPS应激降低了仔猪的生产性能,导致料重比、腹泻率显著提高(P0.05)。试验第14天,日粮中添加1.2%谷氨酰胺显著提高了仔猪的日增重(P0.05),降低了料重比(P0.05)和仔猪腹泻率(P0.05);试验第14天和21天,添加谷氨酰胺对日增重的影响与LPS存在互作效应,即谷氨酰胺缓解了 LPS引起仔猪日增重的下降(P0.05),日粮添加谷氨酰胺对料重比及腹泻率的影响与LPS刺激不存在交互效应。2、LPS应激极显著增加了机体应激指标水平。注射LPS后试验14天断奶仔猪血清中肌酸激酶的水平极显著提高(P0.01),第14天和21天显著增加了血清中皮质醇含量(P0.05)。而日粮中添加1.2%谷氨酰胺后对断奶仔猪血清中肌酸激酶含量极显著降低(P0.01),对第21天试验期葡萄糖的含量极显著提高(P0.01)。谷氨酰胺与LPS应激处理交互作用仅在14日龄对肌酸激酶活力有极显著影响(P0.01),对葡萄糖及皮质醇不存在交互作用。3、日粮中添加1.2%谷氨酰胺可显著提高机体免疫性能,LPS应激后IgA、IgM显著降低(P0.05),谷氨酰胺的添加则使其降低效果得到有效抑制(P0.05)。添加谷氨酰胺显著提高了第14天和21天IFN-y水平(P0.05),缓解LPS应激对IFN-y降低作用(P0.05)。注射LPS导致第14天中的TNF-α水平显著上升,谷氨酰胺的添加对第14天和21天的TNF-α均有明显降低作用(P0.05)。LPS应激显著降低试验第14天补体3水平与第21天补体4水平(P0.05)。谷氨酰胺与LPS交互作用对补体3和补体4作用不显著。4、LPS应激使机体白介素水平提高。注射LPS后断奶仔猪血清中IL-6、IL-8、IL-10显著升高(P0.05),添加谷氨酰胺缓解LPS应激引起的白细胞介素的水平增加。添加谷氨酰胺与LPS应激交互对三种白介素无显著影响。5、LPS应激后对ROS和NO水平明显提高(P0.05),日粮中添加谷氨酰胺,在试验第14天和21天显著提升了 GSH在血清中的含量(P0.05)。谷氨酰胺与LPS应激交互作用对GSH、ROS和NO水平均无显著影响。6、对仔猪的胸腺、脾脏及肠组织粘膜中TLR4、HSP70、MMP-2及MMP-9基因的表达量进行测定,分析结果表明:日粮中添1.2%谷氨酰胺可以显著提高仔猪胸腺、脾脏及肠组织中HSP70基因的表达量(P0.05),显著降低TLR4、MMP-2及MMP-9的表达水平(P0.05)。LPS应激处理组使TLR4、MMP-2及MMP-9基因含量显著升高(P0.05)。基础日粮与LPS应激交互作用对TLR4表达差异显著(P0.05),对HSP70的表达量有极显著性差异(P0.01)。本研究结果表明,日粮中添加谷氨酰胺可增强断奶仔猪生产性能及免疫功能,提高仔猪抗氧化力。谷氨酰胺缓解免疫应激引起的生长抑制与其提高HSP70基因表达,降低TLR4和MMPs基因的表达有关,从而降低炎性细胞因子的分泌,降低机体炎性反应。
[Abstract]:In this paper, we studied the growth performance, immune stress regulation and molecular mechanism of dietary supplementation of glutamine on weanling piglets. 64 weanling piglets were randomly divided into 4 groups, 4 replicates in each group and 4 pigs per repeat. 2 x 2 factorial test was designed, diet type (basal diet or 1.2% glutamine diet) and stress type (injection) LPS or physiological saline). The experimental period 28 days. Group I, group II feeding base diet, respectively in the fourteenth day and 21 days of the experiment, respectively, intraperitoneal injection of normal saline and LPS configuration liquid (100 mu g/kgBW); group III, IV group added 1.2% glutamine in the basal diet, respectively, in Fourteenth days and 21 days of the abdominal injection of normal saline and LPS configuration solution (100 mu g/kg BW). The treatment group took 3h after the day of injection to collect blood from the anterior vena cava, separated the serum, and measured the changes in the serum immunoglobulin and various cytokines. In the twenty-ninth day of the test, the piglet thymus, spleen and intestinal mucosa (duodenum, jejunum, ileum) were slaughtered and the TLR4, HSP70, MMP-2, MMP-9 base in the tissue were detected by fluorescence quantitative PCR The results showed that: 1, LPS stress reduced the production performance of the piglets, resulting in a significant increase in the weight ratio and the diarrhea rate (P0.05). The addition of 1.2% glutamine in the diet increased the daily weight gain (P0.05) of the piglets and reduced the ratio of weight to weight (P0.05). And the diarrhea rate of piglets (P0.05); the effect of adding glutamine on the daily weight gain and the interaction effect of LPS on fourteenth days and 21 days, that is, glutamine relieves the decrease of daily weight gain (P0.05) caused by LPS, and the effect of glutamine on the ratio of feed weight and the rate of diarrhea has no interaction effect with LPS stimulation, and the LPS stress is significantly increased. The level of the body stress index. The level of creatine kinase in the serum of weaned piglets 14 days after the injection of LPS was significantly increased (P0.01). The serum cortisol content (P0.05) was significantly increased in the fourteenth and 21 days. The content of creatine kinase in the serum of weaned piglets was significantly reduced after adding 1.2% glutamine in the diet (P0.01), and for the twenty-first day test period The content of glucose was significantly increased (P0.01). The interaction of glutamine and LPS stress treatment only had a significant effect on the activity of creatine kinase at 14 days of age (P0.01), and there was no interaction between glucose and cortisol.3. The addition of 1.2% glutamine in the diet could significantly improve the immune performance of the body. After LPS stress, IgA, IgM significantly decreased (P0.05), Valley (P0.05). The addition of amides could effectively inhibit the reduction of the effect (P0.05). Adding glutamine significantly increased the IFN-y level of fourteenth and 21 days (P0.05) and alleviated the decrease of IFN-y (P0.05) by LPS stress. The injection of LPS resulted in a significant increase in TNF- alpha in Fourteenth days, and the addition of glutamine to fourteenth days and 21 days of TNF- alpha significantly decreased. The action (P0.05).LPS stress significantly reduced the 3 level of complement fourteenth days and the twenty-first day Complement 4 (P0.05). The interaction of glutamine and LPS had no significant.4 effect on complement 3 and complement 4, and the level of interleukin was increased by LPS stress. IL-6, IL-8, IL-10 in serum of weanling piglets were significantly increased after LPS injection (P0.05), and glutamine was added to alleviate LPS. The interleukin level caused by stress increased. The interaction of glutamine and LPS stress had no significant effect on the three types of interleukin (.5). After LPS stress, the levels of ROS and NO were significantly increased (P0.05), and glutamine was added to the diet. The content of GSH in the blood was significantly increased (P0.05) at the fourteenth and 21 days of the test. The interaction of glutamine and LPS stress The effect on GSH, ROS and NO levels had no significant effect on.6. The expression of TLR4, HSP70, MMP-2 and MMP-9 genes in the mucosa of the thymus, spleen and intestinal tissue of piglets was measured. The results showed that adding 1.2% glutamine in the diet could significantly increase the expression of HSP70 gene in the thymus, spleen and intestinal tissues of piglets (P0.05), significantly reducing TLR4, MMP. The expression level of -2 and MMP-9 (P0.05).LPS stress treatment group increased the content of TLR4, MMP-2 and MMP-9 significantly (P0.05). The interaction between basal diet and LPS stress was significant (P0.05) on TLR4 expression (P0.05), and there was a significant difference in the expression of HSP70. The results of this study showed that the addition of glutamine in the diet could enhance the production of weanling piglets. The performance and immune function can improve the antioxidant power of piglets. The inhibition of growth inhibition by glutamine on immune stress is related to the increase of HSP70 gene expression and the reduction of the expression of TLR4 and MMPs genes, thus reducing the secretion of inflammatory cytokines and reducing the inflammatory response of the body.

【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S828.5

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