7S和11S对仔猪血清IgG亚型及IgE水平和肠黏膜ZO-1表达的影响
发布时间：2018-12-08 19:17
【摘要】：本试验将纯化后的β-伴大豆球蛋白(7S)和大豆球蛋白(11S)以不同剂量添加到断奶仔猪的日粮中,建立仔猪致敏模型。采用ELISA方法检测断奶仔猪血清中的免疫球蛋白,石蜡切片、免疫组织化学的方法检测仔猪小肠黏膜上皮紧密连接蛋白ZO-1的表达与分布,以及实时荧光定量PCR方法研究大豆抗原蛋白对断奶仔猪小肠黏膜上皮紧密连接蛋白ZO-1 mRNA相对表达量的影响,探讨7S和11S对断奶仔猪血清IgG亚型及Ig E和肠粘膜紧密连接蛋白ZO-1表达的影响。选取70头21日龄“杜×长×大”断奶仔猪,随机分成7组,每组10头仔猪。对照组(A组)饲喂基础日粮,β-伴大豆球蛋白(β-conglycinin,7S)B、C、D组和大豆球蛋白(Glycinin,11S)E、F、G组在24 d分别添加500μg·kg-1·W-1、2000μg·kg-1·W-1、5000μg·kg-1·W-1的7S或11S。分别于24、26、28、30日龄采集血液,测定血清中免疫球蛋白含量。试验结束时,每组选取5头仔猪放血致死,采集各段小肠,其中空肠分前、中、后三段采取,置于4%多聚甲醛和液氮中保存,分别用于检测仔猪小肠黏膜上皮紧密连接蛋白ZO-1的表达与分布和检测各段小肠ZO-1 mRNA的相对表达量。试验结果表明:1、大豆抗原蛋白致敏仔猪血清IgG、IgG1、IgG2、IgG3、IgG4和IgE水平升高,其中7S试验组高于11S试验组。2、大豆抗原蛋白致敏仔猪肠黏膜ZO-1的相对表达量降低,且与7S和11S在仔猪日粮中的添加剂量呈负相关;添加剂量相同的7S组ZO-1的相对表达量低于11S组。3、各试验组仔猪肠黏膜紧密连接蛋白ZO-1 mRNA的相对表达量均降低,与7S和11S在仔猪日粮中的添加剂量呈负相关。添加剂量相同的7S组ZO-1 mRNA的相对表达量低于11S组。试验证实,β-伴大豆球蛋白和大豆球蛋对断奶仔猪有明显的致敏作用,β-伴大豆球蛋白致敏性高于大豆球蛋白;β-伴大豆球蛋白和大豆球蛋显著提高致敏仔猪血清中的免疫球蛋白IgG、IgG1、IgG2、IgG3、IgG4和IgE水平,降低肠粘膜上皮紧密连接蛋白ZO-1相对表达量以及ZO-1 mRNA相对表达量。
[Abstract]:In this experiment, purified 尾 -concomitant soybean globulin (7s) and soybean globulin (11s) were added to weaned piglets' diets in different doses to establish sensitizing model of piglets. The expression and distribution of tight junction protein (ZO-1) in small intestinal mucosal epithelium of weaned piglets were detected by ELISA method, paraffin section and immunohistochemical method. The effect of soybean antigen protein on the expression of tight junction protein (ZO-1 mRNA) in small intestinal mucosa of weaned piglets was studied by real-time fluorescence quantitative PCR. To investigate the effects of 7s and 11s on serum IgG subtypes and the expression of Ig E and intestinal mucosal tight junction protein (ZO-1) in weaned piglets. 70 21 day-old weaned piglets were randomly divided into 7 groups with 10 piglets in each group. The control group (group A) was fed with basal diet, 尾 -concomitant soybean globulin (尾-conglycinin,7S) group D and soybean globulin (Glycinin,11S) estradiol. Group G was treated with 渭 g kg-1 W-1G 2000 渭 g kg-1 W-1N 5000 渭 g kg-1 W-1 7s or 11s on the 24th day, respectively. Blood samples were collected at the age of 2830 days, and the content of immunoglobulin in serum was determined. At the end of the experiment, 5 piglets in each group were selected to bleed to death, and the small intestine was collected, in which the jejunum was divided into three sections: anterior, middle and posterior, and stored in 4% paraformaldehyde and liquid nitrogen. It was used to detect the expression and distribution of tight junction protein (ZO-1) in intestinal mucosal epithelium of piglets and to detect the relative expression of ZO-1 mRNA in small intestine. The results showed that: 1. The levels of serum IgG,IgG1,IgG2,IgG3,IgG4 and IgE increased in soybean antigen protein-sensitized piglets, and the relative expression of ZO-1 in intestinal mucosa of 7s group was higher than that of 11s test group, and the relative expression of ZO-1 in intestinal mucosa of soybean antigen protein-sensitized piglets was lower than that of 11s test group. There was a negative correlation between 7s and 11s in the diet of piglets. The relative expression of ZO-1 in 7s group with the same additive content was lower than that in 11s group. 3. The relative expression of ZO-1 mRNA in intestinal mucosal tight junction protein was decreased in all experimental groups, which was negatively correlated with the additive amount of 7s and 11s in diet of piglets. The relative expression of ZO-1 mRNA in 7s group with the same additive content was lower than that in 11s group. The results showed that 尾 -concomitant soybean globulin and soybean ball egg could sensitize weaned piglets, and 尾 -concomitant soybean globulin was more sensitive than soybean globulin. 尾 -concomitant soybean globulin and soybean ball egg significantly increased the levels of immunoglobulin IgG,IgG1,IgG2,IgG3,IgG4 and IgE in serum of sensitized piglets, and decreased the relative expression of ZO-1 and ZO-1 mRNA in intestinal epithelium.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828
本文编号：2368862
[Abstract]:In this experiment, purified 尾 -concomitant soybean globulin (7s) and soybean globulin (11s) were added to weaned piglets' diets in different doses to establish sensitizing model of piglets. The expression and distribution of tight junction protein (ZO-1) in small intestinal mucosal epithelium of weaned piglets were detected by ELISA method, paraffin section and immunohistochemical method. The effect of soybean antigen protein on the expression of tight junction protein (ZO-1 mRNA) in small intestinal mucosa of weaned piglets was studied by real-time fluorescence quantitative PCR. To investigate the effects of 7s and 11s on serum IgG subtypes and the expression of Ig E and intestinal mucosal tight junction protein (ZO-1) in weaned piglets. 70 21 day-old weaned piglets were randomly divided into 7 groups with 10 piglets in each group. The control group (group A) was fed with basal diet, 尾 -concomitant soybean globulin (尾-conglycinin,7S) group D and soybean globulin (Glycinin,11S) estradiol. Group G was treated with 渭 g kg-1 W-1G 2000 渭 g kg-1 W-1N 5000 渭 g kg-1 W-1 7s or 11s on the 24th day, respectively. Blood samples were collected at the age of 2830 days, and the content of immunoglobulin in serum was determined. At the end of the experiment, 5 piglets in each group were selected to bleed to death, and the small intestine was collected, in which the jejunum was divided into three sections: anterior, middle and posterior, and stored in 4% paraformaldehyde and liquid nitrogen. It was used to detect the expression and distribution of tight junction protein (ZO-1) in intestinal mucosal epithelium of piglets and to detect the relative expression of ZO-1 mRNA in small intestine. The results showed that: 1. The levels of serum IgG,IgG1,IgG2,IgG3,IgG4 and IgE increased in soybean antigen protein-sensitized piglets, and the relative expression of ZO-1 in intestinal mucosa of 7s group was higher than that of 11s test group, and the relative expression of ZO-1 in intestinal mucosa of soybean antigen protein-sensitized piglets was lower than that of 11s test group. There was a negative correlation between 7s and 11s in the diet of piglets. The relative expression of ZO-1 in 7s group with the same additive content was lower than that in 11s group. 3. The relative expression of ZO-1 mRNA in intestinal mucosal tight junction protein was decreased in all experimental groups, which was negatively correlated with the additive amount of 7s and 11s in diet of piglets. The relative expression of ZO-1 mRNA in 7s group with the same additive content was lower than that in 11s group. The results showed that 尾 -concomitant soybean globulin and soybean ball egg could sensitize weaned piglets, and 尾 -concomitant soybean globulin was more sensitive than soybean globulin. 尾 -concomitant soybean globulin and soybean ball egg significantly increased the levels of immunoglobulin IgG,IgG1,IgG2,IgG3,IgG4 and IgE in serum of sensitized piglets, and decreased the relative expression of ZO-1 and ZO-1 mRNA in intestinal epithelium.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828
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