本文選題：谷氨酰胺 + 斷奶仔豬��； 參考：《沈陽農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：本論文研究日糧中添加谷氨酰胺對斷奶仔豬生長性能、免疫應(yīng)激調(diào)節(jié)作用及其分子機制。試驗選取64頭斷奶仔豬,隨機分為4組,每組4個重復(fù),每個重復(fù)4頭豬。采用2×2析因試驗設(shè)計,日糧類型(基礎(chǔ)日糧或1.2%谷氨酰胺日糧)和應(yīng)激類型(注射LPS或生理鹽水)。試驗期28天。Ⅰ、Ⅱ組喂食基礎(chǔ)日糧,分別在試驗第14天和21天腹腔注射等量的生理鹽水和LPS配置液(100μg/kgBW);Ⅲ、Ⅳ組在基礎(chǔ)日糧中添加1.2%谷氨酰胺,分別在第14天和21天腹腔注射等量生理鹽水和LPS配置液(100μg/kg BW)。每個處理組當天注射后3h后進行前腔靜脈采血,分離血清,測定血清中免疫球蛋白及各種細胞因子的含量變化。在試驗的第29天屠宰,取仔豬胸腺、脾臟及腸組織黏膜(十二指腸、空腸、回腸),采用熒光定量PCR技術(shù)檢測組織中TLR4、HSP70、MMP-2、MMP-9基因在不同處理組中的表達差異。設(shè)Ⅰ組為本次試驗的對照組,其研究結(jié)果顯示:1、LPS應(yīng)激降低了仔豬的生產(chǎn)性能,導(dǎo)致料重比、腹瀉率顯著提高(P0.05)。試驗第14天,日糧中添加1.2%谷氨酰胺顯著提高了仔豬的日增重(P0.05),降低了料重比(P0.05)和仔豬腹瀉率(P0.05);試驗第14天和21天,添加谷氨酰胺對日增重的影響與LPS存在互作效應(yīng),即谷氨酰胺緩解了 LPS引起仔豬日增重的下降(P0.05),日糧添加谷氨酰胺對料重比及腹瀉率的影響與LPS刺激不存在交互效應(yīng)。2、LPS應(yīng)激極顯著增加了機體應(yīng)激指標水平。注射LPS后試驗14天斷奶仔豬血清中肌酸激酶的水平極顯著提高(P0.01),第14天和21天顯著增加了血清中皮質(zhì)醇含量(P0.05)。而日糧中添加1.2%谷氨酰胺后對斷奶仔豬血清中肌酸激酶含量極顯著降低(P0.01),對第21天試驗期葡萄糖的含量極顯著提高(P0.01)。谷氨酰胺與LPS應(yīng)激處理交互作用僅在14日齡對肌酸激酶活力有極顯著影響(P0.01),對葡萄糖及皮質(zhì)醇不存在交互作用。3、日糧中添加1.2%谷氨酰胺可顯著提高機體免疫性能,LPS應(yīng)激后IgA、IgM顯著降低(P0.05),谷氨酰胺的添加則使其降低效果得到有效抑制(P0.05)。添加谷氨酰胺顯著提高了第14天和21天IFN-y水平(P0.05),緩解LPS應(yīng)激對IFN-y降低作用(P0.05)。注射LPS導(dǎo)致第14天中的TNF-α水平顯著上升,谷氨酰胺的添加對第14天和21天的TNF-α均有明顯降低作用(P0.05)。LPS應(yīng)激顯著降低試驗第14天補體3水平與第21天補體4水平(P0.05)。谷氨酰胺與LPS交互作用對補體3和補體4作用不顯著。4、LPS應(yīng)激使機體白介素水平提高。注射LPS后斷奶仔豬血清中IL-6、IL-8、IL-10顯著升高(P0.05),添加谷氨酰胺緩解LPS應(yīng)激引起的白細胞介素的水平增加。添加谷氨酰胺與LPS應(yīng)激交互對三種白介素無顯著影響。5、LPS應(yīng)激后對ROS和NO水平明顯提高(P0.05),日糧中添加谷氨酰胺,在試驗第14天和21天顯著提升了 GSH在血清中的含量(P0.05)。谷氨酰胺與LPS應(yīng)激交互作用對GSH、ROS和NO水平均無顯著影響。6、對仔豬的胸腺、脾臟及腸組織粘膜中TLR4、HSP70、MMP-2及MMP-9基因的表達量進行測定,分析結(jié)果表明:日糧中添1.2%谷氨酰胺可以顯著提高仔豬胸腺、脾臟及腸組織中HSP70基因的表達量(P0.05),顯著降低TLR4、MMP-2及MMP-9的表達水平(P0.05)。LPS應(yīng)激處理組使TLR4、MMP-2及MMP-9基因含量顯著升高(P0.05)。基礎(chǔ)日糧與LPS應(yīng)激交互作用對TLR4表達差異顯著(P0.05),對HSP70的表達量有極顯著性差異(P0.01)。本研究結(jié)果表明,日糧中添加谷氨酰胺可增強斷奶仔豬生產(chǎn)性能及免疫功能,提高仔豬抗氧化力。谷氨酰胺緩解免疫應(yīng)激引起的生長抑制與其提高HSP70基因表達,降低TLR4和MMPs基因的表達有關(guān),從而降低炎性細胞因子的分泌,降低機體炎性反應(yīng)。
[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.

【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S828.5

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