梅花猪与长白猪仔猪Ghrelin阳性细胞在胃肠道的分布及相关基因表达研究

发布时间：2018-06-27 02:05

  本文选题：梅花猪 + 长白猪　； 参考：《华南农业大学》2016年硕士论文

【摘要】：胃饥饿素(Ghrelin)是一种具有调节食欲、促进生长激素释放、促进胃酸分泌和调节能量稳态等多种生物学功能的胃肠道内分泌激素,主要通过其功能性受体GHSR-1a发挥生物学功能。Ghrelin主要由胃分泌,其分泌细胞数量和分泌活性受发育、环境(日粮和微生物感染)等多种因素的影响,进而影响仔猪的生长发育。梅花猪是广东著名的地方品种,由于瘦肉率低、生长速度慢等特点,正逐渐被市场淘汰,面临灭绝的危机。长白猪有瘦肉率高、生长速度快等优点,广受养猪业的欢迎。为了探究梅花猪和长白猪这两个品种的Ghrelin细胞数量和分泌活性的特点,我们分别对相同日龄、处于相同饲养环境且饲喂相同饲料的梅花猪和长白猪仔猪胃肠道各个部位的Ghrelin细胞数量以及Ghrelin的表达进行研究。本试验选取30日龄断奶的纯系梅花猪和丹系长白猪的仔猪为研究对象,在相同条件(日龄、日粮、饲养环境相同)下饲养至65日龄,采集血清样品后屠宰,然后采集胃肠道各部位组织样品。应用免疫组织化学和免疫荧光的方法对梅花猪和长白猪的胃肠道各部位的Ghrelin细胞进行鉴定和计数,应用ELISA的方法检测上述两个品种仔猪胃肠道组织和血清Ghrelin含量,应用荧光定量PCR的方法测定上述两个品种仔猪胃肠道各部位粘膜组织的GHRL、GHSR和GHSR-1a的基因表达。研究结果如下:(1)免疫组织化学和免疫荧光结果表明,梅花猪和长白猪仔猪的胃腺体部、胃幽门、十二指肠和空肠都存在Ghrelin细胞,且Ghrelin细胞分为开放型和封闭型两种。(2)长白猪胃幽门的Ghrelin阳性细胞密度显著高于梅花猪,梅花猪空肠的阳性细胞密度极显著高于长白猪。梅花猪空肠的闭合型Ghrelin阳性细胞密度极显著高于长白猪。长白猪胃幽门和十二指肠的开放型Ghrelin阳性细胞密度分别极显著和显著高于梅花猪,梅花猪空肠的开放型阳性细胞密度极显著高于长白猪。(3)梅花猪与长白猪Ghrelin细胞数量分布趋势一致,都是沿肠道方向递减。(4)梅花猪在胃腺体部、胃幽门、十二指肠和空肠中闭合型和开放型Ghrelin阳性细胞的比例分布为9:1、6:1、4:1和3:1;而长白猪在胃腺体部、胃幽门、十二指肠和空肠中闭合型与开放型阳性细胞的比例分别为9:1、7:3、2:1和7:2。(5)长白猪仔猪胃腺体部的Ghrelin浓度极显著高于梅花猪,梅花猪血清、十二指肠和空肠的Ghrelin浓度极显著高于长白猪。(6)梅花猪胃腺体部的GHRL表达水平显著高于长白猪。梅花猪与长白猪GHRL表达水平趋势一致,与Ghrelin细胞数量分布趋势一致,都是沿肠道方向递减。(7)长白猪胃腺体部、胃幽门和空肠的GHSR的表达水平显著高于梅花猪。长白猪空肠的GHSR-1a的表达水平显著高于梅花猪。
[Abstract]:Ghrelin is a kind of gastrointestinal endocrine hormone which can regulate appetite, promote the release of growth hormone, promote gastric acid secretion and regulate energy homeostasis. Ghrelin is mainly secreted by stomach through its functional receptor GHSR-1a. The number and activity of secreting cells are affected by many factors, such as development, environment (diet and microbial infection), and then affect the growth and development of piglets. Meihua pig is a famous local breed in Guangdong province. Because of its low lean meat rate and slow growth rate, it is gradually eliminated by the market and is facing the crisis of extinction. Landrace pig has the advantages of high lean meat rate and fast growth rate, and is widely welcomed by pig industry. In order to investigate the characteristics of ghrelin cell number and secretory activity in two breeds of Plum Flower and Landrace, we studied the characteristics of the same age at the same age. The number of ghrelin cells and the expression of ghrelin in different parts of gastrointestinal tract of Plum and Landrace piglets were studied in the same feeding environment. In this experiment, pure-line plum blossom pigs weaned at 30 days of age and piglets of Landrace Landrace piglets were selected as study objects. The piglets were raised to 65 days of age under the same conditions (day age, diet, feeding environment), and serum samples were collected and slaughtered. Then the samples were collected from various parts of the gastrointestinal tract. Ghrelin cells were identified and counted in various parts of the gastrointestinal tract of piggy and Landrace pigs by immunohistochemistry and immunofluorescence. The contents of ghrelin in gastrointestinal tissue and serum of these two breeds of piglets were detected by Elisa. The gene expression of GHRLGHSR and GHSR-1a in different parts of gastrointestinal mucosa of the two piglets were detected by fluorescence quantitative PCR (FQ-PCR). The results were as follows: (1) Immunohistochemistry and immunofluorescence showed that ghrelin cells were present in gastric glands, pylorus, duodenum and jejunum of piggy and Landrace piglets. Ghrelin cells were divided into open type and closed type. (2) the density of ghrelin positive cells in stomach pylorus of Landrace pig was significantly higher than that in plum flower pig, and the density of positive cell in jejunum of plum white pig was significantly higher than that of Landrace pig. The density of closed ghrelin positive cells in jejunum of piggy pigs was significantly higher than that of Landrace pigs. The densities of open ghrelin positive cells in stomach pylorus and duodenum of Landrace pigs were significantly higher than those in piggy pigs. The density of open positive cells in jejunum of piggy pigs was significantly higher than that of Landrace pigs. (3) the distribution of ghrelin cells in Plum and Landrace pigs was the same and decreased along the direction of intestine. The percentages of closed and open ghrelin positive cells in duodenum and jejunum were 9: 1, 6: 1, 4: 1 and 3: 1, respectively. The ratios of closed and open positive cells in duodenum and jejunum were 9: 1: 7: 3: 1 and 7: 2: 1 respectively. (5) ghrelin concentration in stomach gland of Landrace piglets was significantly higher than that in piggy pigs and serum of piggy pigs. Ghrelin concentration in duodenum and jejunum was significantly higher than that in Landrace pig. (6) the expression of GHRL in stomach gland of piggy pigs was significantly higher than that in Landrace pigs. The expression level of GHRL was the same in Pigs and Landrace pigs, and the distribution of GHRL cells was consistent with that of ghrelin cells. (7) the expression of GHSR in stomach gland, pylorus and jejunum of Landrace pigs was significantly higher than that in Pigs. The expression level of GHSR-1a in jejunum of Landrace pig was significantly higher than that in piggy pig.
【学位授予单位】：华南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S828
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本文编号：2072255