姜黄素对大肠杆菌攻毒仔猪生长性能及肠粘膜屏障功能的影响

发布时间：2018-05-31 04:44

本文选题：姜黄素 + 仔猪　；参考：《海南大学》2015年硕士论文

【摘要】：为探讨日粮中添加姜黄素对断奶仔猪生长性能、抗氧化性能、肠道机械屏障与免疫屏障的影响,和其替代抗生素应用于饲料添加剂的可行性,并探索适合的添加量。本试验选用50头21日龄断奶仔猪,随机分为5组,每组10个重复,每个重复1头猪。预试期采用大肠杆菌进行攻毒(1×109个细胞/头),其中对照组饲喂基础日粮,试验组在基础日粮中分别添加50mg/kg喹烯酮,200mg/kg、300mg/kg和400mg/kg姜黄素,试验期21d。 1为探讨姜黄素对断奶仔猪生长性能和抗氧化性能的影响,实验检测了仔猪平均日增重(ADG)、平均日采食量(ADFI)及料重比(F/G),并对仔猪血清谷胱甘肽过氧化酶(GSH-PX)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)的活性、总抗氧化能力(T-AOC)和丙二醛(MDA)含量进行检测,结果发现：与对照组相比,日粮中添加50mg/kg喹烯酮、300或400ng/kg姜黄素可以显著降低断奶仔猪的料重比(P0.05)；喹烯酮组GSH-PX、CAT、SOD活性显著升高(P＜0.05),MDA含量显著降低(P0.05),300mg/kg和400mg/kg姜黄素组断奶仔猪血清中GSH-PX、CAT、SOD活性和T-AOC显著升高(P0.05),400mg/kg姜黄素组MDA含量显著降低(P0.05)。结果表明,日粮中添加喹烯酮、300mg/kg或400mg/kg姜黄素能促进仔猪生长,提高仔猪抗氧化性能。 2为探讨姜黄素对仔猪肠粘膜机械屏障的影响,在光镜下测量空肠、回肠绒毛高度、宽度及隐窝深度,计算绒腺比,采用ELISA法测定DAO活性和D-乳酸的含量,采用免疫组化和荧光定量PCR技术测定了Claudin-1、Occludin和ZO-1mRNA及蛋白表达。结果表明：1)日粮中添加300mg/kg或400mg/kg姜黄素显著提高空肠和回肠绒毛高度(P0.05)与绒腺比(P0.05),降低绒毛宽度(P0.05)和隐窝深度(P0.05),喹烯酮组空肠与回肠绒毛宽度和空肠隐窝深度显著降低(P0.05),绒腺比显著升高(P0.05)；2)300和400mg/kg姜黄素组可显著降低血浆中DAO活性(P0.05)和D-乳酸含量(P0.05),200mg/kg和喹烯酮组血浆DAO和D-乳酸与对照组差异不显著(P0.05)；3)300和400]mg/kg姜黄素组Occludin和ZO-1蛋白表达升高(P0.05), Claudin-1蛋白表达无明显差异(P0.05),喹烯酮和200mg/kg姜黄素组Claudin-1、Occludin和ZO-1蛋白表达量与对照组差异不显著(P0.05)；4)300mg/kg和400mg/kg姜黄素组Occludin和ZO-1mRNA相对表达量显著升高(P0.05),400mg/kg姜黄素组Claudin-1mRNA相对表达量显著升高(P0.05),200mg/kg姜黄素组ZO-1mRNA表达量显著高于对照组(P0.05),喹烯酮组Claudin-1、Occludin和ZO-1mRNA相对表达量与对照组差异不显著(P0.05),结果显示：日粮中添加300mg/kg或400mg/kg姜黄素可降低肠粘膜通透性,提高紧密连接蛋白及其mRNA表达,进而改善肠粘膜形态结构保护肠粘膜机械屏障。 3为探讨姜黄素对仔猪肠粘膜免疫屏障的影响,试验采用PAS染色法统计空肠、回肠粘膜杯状细胞数,应用免疫组化法测定sIgA蛋白的表达,利用荧光定量PCR方法测定了空肠粘膜IL-1β、1L-6、 IL-10、 TNF-a和TLR4mRNA的相对表达量。结果显示：日粮中添加300mg/kg或400mg/kg姜黄素可显著提高空肠和回肠杯状细胞数量(P0.05)；降低TLR4和Calprotectin蛋白的表达(P0.05),提高sIgA蛋白表达,降低IL-1β、1L-6、TNF-α和TLR4mRNA的相对表达量(P0.05),提高IL-10mRNA的相对表达量(P0.05)；喹烯酮组回肠杯状细胞数量显著增加(P0.05),显著降低空肠TLR4mRNA口蛋白表达(P0.05)；200mg/kg姜黄素组显著增加回肠杯状细胞数量(P0.05)。结果显示：添加300mg/kg或400mg/kg姜黄素可抑制肠粘膜炎症反应的发生,提高肠粘膜免疫屏障功能。综上所述,在日粮中添加300mg/kg或400mg/kg姜黄素可以促进仔猪生长,提高仔猪抗氧化性,改善肠粘膜形态结构,降低肠粘膜通透性,提高肠粘膜紧密连接和免疫功能,从而起到了保护肠道屏障的作用。因此得出结论,姜黄素具有作为绿色饲料添加剂替代抗生素的潜力。
[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.
【学位授予单位】：海南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828.5

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