發(fā)布時間：2018-05-31 04:44

  本文選題：姜黃素 + 仔豬��； 參考：《海南大學》2015年碩士論文

【摘要】：為探討日糧中添加姜黃素對斷奶仔豬生長性能、抗氧化性能、腸道機械屏障與免疫屏障的影響,和其替代抗生素應用于飼料添加劑的可行性,并探索適合的添加量。本試驗選用50頭21日齡斷奶仔豬,隨機分為5組,每組10個重復,每個重復1頭豬。預試期采用大腸桿菌進行攻毒(1×109個細胞/頭),其中對照組飼喂基礎日糧,試驗組在基礎日糧中分別添加50mg/kg喹烯酮,200mg/kg、300mg/kg和400mg/kg姜黃素,試驗期21d。 1為探討姜黃素對斷奶仔豬生長性能和抗氧化性能的影響,實驗檢測了仔豬平均日增重(ADG)、平均日采食量(ADFI)及料重比(F/G),并對仔豬血清谷胱甘肽過氧化酶(GSH-PX)、過氧化氫酶(CAT)、超氧化物歧化酶(SOD)的活性、總抗氧化能力(T-AOC)和丙二醛(MDA)含量進行檢測,結果發(fā)現(xiàn)：與對照組相比,日糧中添加50mg/kg喹烯酮、300或400ng/kg姜黃素可以顯著降低斷奶仔豬的料重比(P0.05)；喹烯酮組GSH-PX、CAT、SOD活性顯著升高(P＜0.05),MDA含量顯著降低(P0.05),300mg/kg和400mg/kg姜黃素組斷奶仔豬血清中GSH-PX、CAT、SOD活性和T-AOC顯著升高(P0.05),400mg/kg姜黃素組MDA含量顯著降低(P0.05)。結果表明,日糧中添加喹烯酮、300mg/kg或400mg/kg姜黃素能促進仔豬生長,提高仔豬抗氧化性能。 2為探討姜黃素對仔豬腸粘膜機械屏障的影響,在光鏡下測量空腸、回腸絨毛高度、寬度及隱窩深度,計算絨腺比,采用ELISA法測定DAO活性和D-乳酸的含量,采用免疫組化和熒光定量PCR技術測定了Claudin-1、Occludin和ZO-1mRNA及蛋白表達。結果表明：1)日糧中添加300mg/kg或400mg/kg姜黃素顯著提高空腸和回腸絨毛高度(P0.05)與絨腺比(P0.05),降低絨毛寬度(P0.05)和隱窩深度(P0.05),喹烯酮組空腸與回腸絨毛寬度和空腸隱窩深度顯著降低(P0.05),絨腺比顯著升高(P0.05)；2)300和400mg/kg姜黃素組可顯著降低血漿中DAO活性(P0.05)和D-乳酸含量(P0.05),200mg/kg和喹烯酮組血漿DAO和D-乳酸與對照組差異不顯著(P0.05)；3)300和400]mg/kg姜黃素組Occludin和ZO-1蛋白表達升高(P0.05), Claudin-1蛋白表達無明顯差異(P0.05),喹烯酮和200mg/kg姜黃素組Claudin-1、Occludin和ZO-1蛋白表達量與對照組差異不顯著(P0.05)；4)300mg/kg和400mg/kg姜黃素組Occludin和ZO-1mRNA相對表達量顯著升高(P0.05),400mg/kg姜黃素組Claudin-1mRNA相對表達量顯著升高(P0.05),200mg/kg姜黃素組ZO-1mRNA表達量顯著高于對照組(P0.05),喹烯酮組Claudin-1、Occludin和ZO-1mRNA相對表達量與對照組差異不顯著(P0.05),結果顯示：日糧中添加300mg/kg或400mg/kg姜黃素可降低腸粘膜通透性,提高緊密連接蛋白及其mRNA表達,進而改善腸粘膜形態(tài)結構保護腸粘膜機械屏障。 3為探討姜黃素對仔豬腸粘膜免疫屏障的影響,試驗采用PAS染色法統(tǒng)計空腸、回腸粘膜杯狀細胞數(shù),應用免疫組化法測定sIgA蛋白的表達,利用熒光定量PCR方法測定了空腸粘膜IL-1β、1L-6、 IL-10、 TNF-a和TLR4mRNA的相對表達量。結果顯示：日糧中添加300mg/kg或400mg/kg姜黃素可顯著提高空腸和回腸杯狀細胞數(shù)量(P0.05)；降低TLR4和Calprotectin蛋白的表達(P0.05),提高sIgA蛋白表達,降低IL-1β、1L-6、TNF-α和TLR4mRNA的相對表達量(P0.05),提高IL-10mRNA的相對表達量(P0.05)；喹烯酮組回腸杯狀細胞數(shù)量顯著增加(P0.05),顯著降低空腸TLR4mRNA口蛋白表達(P0.05)；200mg/kg姜黃素組顯著增加回腸杯狀細胞數(shù)量(P0.05)。結果顯示：添加300mg/kg或400mg/kg姜黃素可抑制腸粘膜炎癥反應的發(fā)生,提高腸粘膜免疫屏障功能。 綜上所述,在日糧中添加300mg/kg或400mg/kg姜黃素可以促進仔豬生長,提高仔豬抗氧化性,改善腸粘膜形態(tài)結構,降低腸粘膜通透性,提高腸粘膜緊密連接和免疫功能,從而起到了保護腸道屏障的作用。因此得出結論,姜黃素具有作為綠色飼料添加劑替代抗生素的潛力。
[Abstract]:In order to explore the effects of curcumin on growth performance, antioxidant properties, intestinal mechanical barrier and immune barrier, and the feasibility of applying antibiotics to feed additives, 50 21 day old weanling piglets were randomly divided into 5 groups, 10 repetitions in each group, and 1 replicates in each group. In the pre test period, Escherichia coli was used to attack the virus (1 x 109 cells / head), and the control group was fed with basal diet. The test group added 50mg/kg, 200mg/kg, 300mg/kg and 400mg/kg curcumin in the basal diet, and the test period was 21d.
1 in order to investigate the effects of curcumin on growth performance and antioxidant activity of weaned piglets, the average daily gain (ADG), average daily feed intake (ADFI) and material weight ratio (F/G) were tested, and the activity of glutathione peroxidase (GSH-PX), catalase (CAT), superoxide dismutase (SOD), total antioxidant capacity (T-AOC) and C in piglets were also investigated. The content of two aldehyde (MDA) was detected. The results were as follows: compared with the control group, the addition of 50mg/kg and 400ng/kg curcumin in the diet could significantly reduce the feed weight ratio (P0.05) of weanling piglets; the GSH-PX, CAT, SOD activity of the Q group was significantly increased (P < 0.05), and the MDA content was significantly reduced (P0.05), 300mg/kg and 400mg/kg curcumin group weaned piglets. The activity of GSH-PX, CAT, SOD and T-AOC in the serum increased significantly (P0.05), and the MDA content in the 400mg/kg curcumin group decreased significantly (P0.05). The results showed that the addition of CAT and curcumin in the diet could promote the growth of piglets and improve the antioxidant performance of piglets.
2 in order to investigate the effect of curcumin on the intestinal mucosal mechanical barrier of piglets, we measured the jejunum, the height of the ileum, the width and the depth of the recess in the light microscope, calculated the ratio of the cashmere glands, and measured the activity of DAO and the content of D- lactic acid by ELISA. The expression of Claudin-1, Occludin, ZO-1mRNA and protein were measured by immunohistochemistry and fluorescence quantitative PCR. The results showed that the expression of Claudin-1, ZO-1mRNA and protein were determined. 1) the addition of 300mg/kg or 400mg/kg curcumin in the diet increased the height of jejunum and ileum (P0.05) and the ratio of cashmere glands (P0.05), reduced the width of villi (P0.05) and the depth of the recess (P0.05). The width of jejunum and the ileum and the depth of the jejunum recess decreased significantly (P0.05), the ratio of the cashmere gland was significantly increased (P0.05), and 2) 300 and 400mg/kg curcumin. DAO activity (P0.05) and D- lactate content in plasma were significantly reduced (P0.05). There was no significant difference between the plasma DAO and D- lactate in 200mg/kg and the group of 200mg/kg and the control group (P0.05); 3) the expression of Occludin and ZO-1 protein in the 300 and 400]mg/kg curcumin group increased (P0.05). The expression of din-1, Occludin and ZO-1 protein was not significantly different from that of the control group (P0.05); 4) the relative expression of Occludin and ZO-1mRNA in 300mg/kg and 400mg/kg curcumin group increased significantly (P0.05), and Claudin-1mRNA relative expression in the 400mg/kg curcumin group increased significantly (P0.05), and the expression of Curcumin group was significantly higher than that of the control group. The relative expression of Claudin-1, Occludin and ZO-1mRNA in the ketone group was not significantly different from that of the control group (P0.05). The results showed that the addition of 300mg/kg or 400mg/kg curcumin in the diet could reduce the permeability of intestinal mucosa, improve the expression of tight connexin and its mRNA, and then improve the intestinal mucosal structure and protect the intestinal mucosal mechanical barrier.
3 in order to investigate the effect of curcumin on the intestinal mucosal immune barrier of piglets, PAS staining was used to count the jejunum, the number of goblet cells in the ileum and the expression of sIgA protein by immunohistochemical method. The relative expression of IL-1 beta, 1L-6, IL-10, TNF-a and TLR4mRNA in the jejunum mucosa was measured by fluorescence quantitative PCR. The results showed: in the diet Adding 300mg/kg or 400mg/kg curcumin could significantly increase the number of goblet cells in jejunum and ileum (P0.05), decrease the expression of TLR4 and Calprotectin protein (P0.05), increase the expression of sIgA protein, decrease the relative expression of IL-1 beta, 1L-6, TNF- A and TLR4mRNA (P0.05), and increase the relative expression of sIgA, and the ileum goblet cells in the O3 group The number of TLR4mRNA (P0.05) decreased significantly (P0.05), and the number of 200mg/kg curcumin significantly increased the number of ileum goblet cells (P0.05). The results showed that the addition of 300mg/kg or 400mg/kg curcumin could inhibit the occurrence of intestinal mucosal inflammatory reaction and improve the intestinal mucosal immune barrier function.
To sum up, adding 300mg/kg or 400mg/kg curcumin to the diet can promote the growth of piglets, improve the antioxidant activity of the piglets, improve the morphological structure of the intestinal mucosa, reduce the permeability of the intestinal mucosa, improve the close connection and immune function of the intestinal mucosa, and thus protect the intestinal barrier. Therefore, it is concluded that curcumin can be used as green feeding. The potential of material additives to replace antibiotics.
【學位授予單位】：海南大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S828.5

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