异戊酸对犊牛小肠消化酶活性、黏膜组织形态及其功能基因表达的影响
发布时间：2019-04-23 12:39
【摘要】：试验选用36头发育正常、体重45.09±0.90kg的15日龄哺乳犊牛,随机分为4组,每组9头,测定日粮添加异戊酸对犊牛小肠生长发育、小肠消化酶活性变化及其小肠功能基因表达的影响。所有犊牛在45日龄断奶,对照组饲喂基础日粮,试验组分别在基础日粮基础上添加异戊酸3、6和9g/d。分别于30日龄、60日龄、90日龄从各组随机抽取3头,晨饲前称重后进行屠宰采样。采集小肠4个不同部位(十二指肠、空肠近端1/4处、空肠远端1/4处和回肠末端)的肠一份用于小肠消化酶的测定;小肠黏膜组织样品2份,一份用于形态学观察及扫描电镜观察,另一份通过实时荧光定量技术检测生长激素受体(GHR)、胰岛素受体(INSR)、胰岛素样生长因子-1(IGF-1)、寡肽共转运载体1(PepT1)及钠-葡萄糖共转运载体1 (SGLT1)在小肠黏膜不同部位的基因表达。结果表明： (1)60和90日龄犊牛,6g/d、9g/d异戊酸添加组犊牛的十二指肠、空肠、回肠和小肠总长度都显著增加；(2)各日龄犊牛,6g/d和9g/d异戊酸添加组犊牛的空肠近端和空肠远端的肠壁厚度、肠黏膜厚度、肠绒毛长度都显著增加,绒毛长度/隐窝深度的比值(VH/CH)有增加的趋势; (3)各日龄犊牛,6g/d和9g/d异戊酸添加组犊牛的空肠近端、空肠远端和回肠乳糖酶、淀粉酶、蛋白酶及脂肪酶活性都显著增加,而且异戊酸对于十二指肠各种消化酶活性也都呈现促进作用； (4)30日龄犊牛,6g/d、9g/d异戊酸添加组犊牛的十二指肠、空肠近端、空肠远端及回肠黏膜的生长激素受体mRNA (GHR mRNA)、胰岛素受体mRNA(INSR mRNA)、胰岛素样生长因子-1 mRNA (IGF-1 mRNA)和钠-葡萄糖共转运载体1 mRNA (SGLT1 mRNA)的表达量都显著增加；而60日龄犊牛,GHR mRNA、INSR mRNA、IGF-1 mRNA、SGLT1 mRNA以及寡肽共转运载体1 mRNA (Pep T1 mRNA)的表达量在空肠近端和空肠远端都显著增加,此外,异戊酸对于GHR mRNA、INSR mRNA、IGF-1 mRNA、SGLT1 mRNA和Pep T1 mRNA在十二指肠和回肠的表达都表现有显著促进作用。综上,可做推断,日粮添加异戌酸对犊牛小肠的生长发育、小肠消化酶活性及小肠功能基因的表达都具有显著促进作用,且最适添加量为6g/d。
[Abstract]:Thirty-six 15-day-old lactating calves with normal development and weighing 45.09 卤0.90kg were randomly divided into 4 groups, 9 in each group. The effects of isovalproic acid on the growth and development of small intestine in calves were measured. Changes of digestive enzyme activity in small intestine and its effect on gene expression of intestinal function. All calves were weaned at the age of 45 days. The control group was fed with basal diet and the experimental group was fed with isovalproic acid 3, 6 and 9 g / d on the basis of basal diet, respectively. At the age of 30 days, 60 days and 90 days, 3 heads were randomly sampled from each group, and then slaughtered and sampled before morning feeding. One piece of intestine from 4 different parts of small intestine (duodenum, proximal jejunum, distal jejunum and ileum) was collected and used for the determination of digestive enzymes in small intestine. Two small intestinal mucosa tissue samples, one for morphological observation and scanning electron microscope observation, and the other for detecting growth hormone receptor (GHR), insulin receptor (INSR), by real-time fluorescence quantitative technique. Gene expression of insulin-like growth factor-1 (IGF-1), oligopeptide co-transport vector 1 (PepT1) and sodium-glucose co-transport vector 1 (SGLT1) in small intestinal mucosa. The results showed that: (1) the total length of duodenum, jejunum, ileum and small intestine of 60-and 90-day-old calves, 6 g / d, 9 g / d isopentanoic acid-supplemented calves were significantly increased; (2) the thickness of proximal jejunum and distal jejunum, the thickness of intestinal mucosa and the length of intestinal villus of calves fed with 6g/d and 9g/d isovalerate were significantly increased. The ratio of villus length to recess depth (VH/CH) increased. (3) the activities of lactase, amylase, protease and lipase in the proximal jejunum, distal jejunum and ileum of calves fed with 6g/d and 9g/d isovalerate were significantly increased. Isovalerate also promoted the activities of digestive enzymes in duodenum. (4) growth hormone receptor mRNA (GHR mRNA), insulin receptor mRNA (INSR mRNA), in duodenum, proximal jejunum, distal jejunum and ileum mucosa of 30-day-old calves, 6 g / d, 9 g / kg isopentanoic acid group. The expression of insulin-like growth factor-1 mRNA (IGF-1 mRNA) and sodium-glucose co-transport vector 1 mRNA (SGLT1 mRNA) were significantly increased. However, the expression of GHR mRNA,INSR mRNA,IGF-1 mRNA,SGLT1 mRNA and 1 mRNA (Pep T1 mRNA in 60 day old calves increased significantly both in proximal jejunum and distal jejunum. In addition, isopentanoic acid could significantly increase the expression level of GHR mRNA,INSR mRNA,IGF-1 mRNA, in the proximal jejunum and distal jejunum. The expression of SGLT1 mRNA and Pep T1 mRNA in duodenum and ileum were significantly enhanced. In conclusion, it can be inferred that diet supplemented with isovalproic acid can significantly promote the growth and development of the small intestine, the activity of digestive enzymes and the expression of small intestinal function genes in calves, and the optimum dosage is 6 g / d.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823.5
本文编号：2463476
[Abstract]:Thirty-six 15-day-old lactating calves with normal development and weighing 45.09 卤0.90kg were randomly divided into 4 groups, 9 in each group. The effects of isovalproic acid on the growth and development of small intestine in calves were measured. Changes of digestive enzyme activity in small intestine and its effect on gene expression of intestinal function. All calves were weaned at the age of 45 days. The control group was fed with basal diet and the experimental group was fed with isovalproic acid 3, 6 and 9 g / d on the basis of basal diet, respectively. At the age of 30 days, 60 days and 90 days, 3 heads were randomly sampled from each group, and then slaughtered and sampled before morning feeding. One piece of intestine from 4 different parts of small intestine (duodenum, proximal jejunum, distal jejunum and ileum) was collected and used for the determination of digestive enzymes in small intestine. Two small intestinal mucosa tissue samples, one for morphological observation and scanning electron microscope observation, and the other for detecting growth hormone receptor (GHR), insulin receptor (INSR), by real-time fluorescence quantitative technique. Gene expression of insulin-like growth factor-1 (IGF-1), oligopeptide co-transport vector 1 (PepT1) and sodium-glucose co-transport vector 1 (SGLT1) in small intestinal mucosa. The results showed that: (1) the total length of duodenum, jejunum, ileum and small intestine of 60-and 90-day-old calves, 6 g / d, 9 g / d isopentanoic acid-supplemented calves were significantly increased; (2) the thickness of proximal jejunum and distal jejunum, the thickness of intestinal mucosa and the length of intestinal villus of calves fed with 6g/d and 9g/d isovalerate were significantly increased. The ratio of villus length to recess depth (VH/CH) increased. (3) the activities of lactase, amylase, protease and lipase in the proximal jejunum, distal jejunum and ileum of calves fed with 6g/d and 9g/d isovalerate were significantly increased. Isovalerate also promoted the activities of digestive enzymes in duodenum. (4) growth hormone receptor mRNA (GHR mRNA), insulin receptor mRNA (INSR mRNA), in duodenum, proximal jejunum, distal jejunum and ileum mucosa of 30-day-old calves, 6 g / d, 9 g / kg isopentanoic acid group. The expression of insulin-like growth factor-1 mRNA (IGF-1 mRNA) and sodium-glucose co-transport vector 1 mRNA (SGLT1 mRNA) were significantly increased. However, the expression of GHR mRNA,INSR mRNA,IGF-1 mRNA,SGLT1 mRNA and 1 mRNA (Pep T1 mRNA in 60 day old calves increased significantly both in proximal jejunum and distal jejunum. In addition, isopentanoic acid could significantly increase the expression level of GHR mRNA,INSR mRNA,IGF-1 mRNA, in the proximal jejunum and distal jejunum. The expression of SGLT1 mRNA and Pep T1 mRNA in duodenum and ileum were significantly enhanced. In conclusion, it can be inferred that diet supplemented with isovalproic acid can significantly promote the growth and development of the small intestine, the activity of digestive enzymes and the expression of small intestinal function genes in calves, and the optimum dosage is 6 g / d.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823.5
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