β-伴大豆球蛋白及其主要抗原表位与大鼠小肠上皮黏膜蛋白结合的比较
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摘要
本试验旨在比较β-伴大豆球蛋白及其主要抗原表位与大鼠小肠上皮黏膜蛋白的结合情况。试验选取12只SD大鼠,随机分为2组(对照组和致敏组),每组6个重复,每个重复1只。对照组和致敏组均饲喂不含任何豆科成分来源的饲粮,致敏组以β-伴大豆球蛋白为致敏源进行攻击和激发,并采用免疫组织化学法对β-伴大豆球蛋白及其主要抗原表位与小肠上皮黏膜蛋白的结合蛋白进行检测。结果表明:1)对照组的空肠前段β-伴大豆球蛋白结合蛋白的含量与致敏组存在显著性差异(P<0.05)。对照组的十二指肠和空肠前段β-伴大豆球蛋白结合蛋白的含量显著高于空肠中段、空肠后段和回肠(P<0.05),致敏组的十二指肠β-伴大豆球蛋白结合蛋白的含量显著高于空肠前段、空肠中段、空肠后段和回肠(P<0.05)。2)对照组的空肠后段和回肠α2抗原表位结合蛋白的含量与致敏组存在显著性差异(P<0.05)。对照组的十二指肠和空肠前段α2抗原表位结合蛋白的含量显著高于空肠中段、空肠后段和回肠(P<0.05);致敏组的十二指肠α2抗原表位结合蛋白的含量显著高于空肠前段、空肠中段、空肠后段和回肠(P<0.05)。3)对照组的十二指肠和空肠后段β4抗原表位结合蛋白的含量与对照组存在显著性差异(P<0.05)。对照组和致敏组的十二指肠、空肠前段和空肠中段β4抗原表位结合蛋白的含量显著高于空肠后段和回肠(P<0.05)。由此可见,十二指肠β-伴大豆球蛋白、α2和β4抗原表位结合蛋白的含量最多,空肠后段和回肠较少。致敏组β-伴大豆球蛋白、α2和β4抗原表位结合蛋白的含量普遍低于对照组。
This experiment was conducted to compare the situation of β-conglycinin and its major antigen epitope combined with small intestinal epithelial mucosal proteins in rats. Twelve SD rats were selected and randomly divided into 2 groups with 6 replicates per group and 1 rat per replicate. Both the control group and sensitization group were fed a diet without leguminous component sources,rats in the sensitization group were attack and stimulate using β-conglycinin as sensitizer,and the immunohistochemical method was used to detect the protein of β-conglycinin and its major antigen epitope combined with binding protein in intestinal epithelial mucosal protein. The results showed as follows: 1) there was a significant difference on content of β-conglycinin binding protein in jejunum anterior segment between control group and sensitization group( P<0.05). The content of β-conglycinin binding protein in duodenum and jejunum anterior segment of the control group was significantly higher than that in jejunum middle segment,jejunum posterior segment and ileum( P<0.05),the content of β-conglycinin binding protein in duodenum of the sensitization group was significantly higher than that in jejunum anterior segment,jejunum middle segment,jejunum posterior segment and ileum( P < 0.05).2) There was a significant difference on content of α2 antigen epitope binding protein in jejunum posterior segment and ileum between control group and sensitization group( P < 0. 05). The content of α2 antigen epitope binding protein in duodenum and jejunum anterior segment of the control group was significantly higher than that in jejunum middle segment,jejunum posterior segment and ileum( P < 0. 05),the content of α2 antigen epitope binding protein in duodenum of the sensitization group was significantly higher than that in and jejunum anterior segment,jejunum middle segment,jejunum posterior segment and ileum( P<0.05). 3) There was a significant difference on content of β4 antigen epitope binding protein in duodenum and jejunum posterior segment between control group and sensitization group( P<0.05). The content of β4 antigen epitope binding protein in duodenum and jejunum anterior segment and jejunum middle segment of control group and sensitization group was significantly higher than that jejunum posterior segment and ileum( P < 0. 05). In conclusion,the content of β-conglycinin,α2 and β4 antigen epitope binding protein in duodenum is the most,and that in jejunum posterior segment and ileum is less. The content of β-conglycinin,α2 and β4 antigen epitope binding protein of sensitization group is generally lower than that of the control group.[Chinese Journal of Animal Nutrition,2019,31(7):3235-3241]
This experiment was conducted to compare the situation of β-conglycinin and its major antigen epitope combined with small intestinal epithelial mucosal proteins in rats. Twelve SD rats were selected and randomly divided into 2 groups with 6 replicates per group and 1 rat per replicate. Both the control group and sensitization group were fed a diet without leguminous component sources,rats in the sensitization group were attack and stimulate using β-conglycinin as sensitizer,and the immunohistochemical method was used to detect the protein of β-conglycinin and its major antigen epitope combined with binding protein in intestinal epithelial mucosal protein. The results showed as follows: 1) there was a significant difference on content of β-conglycinin binding protein in jejunum anterior segment between control group and sensitization group( P<0.05). The content of β-conglycinin binding protein in duodenum and jejunum anterior segment of the control group was significantly higher than that in jejunum middle segment,jejunum posterior segment and ileum( P<0.05),the content of β-conglycinin binding protein in duodenum of the sensitization group was significantly higher than that in jejunum anterior segment,jejunum middle segment,jejunum posterior segment and ileum( P < 0.05).2) There was a significant difference on content of α2 antigen epitope binding protein in jejunum posterior segment and ileum between control group and sensitization group( P < 0. 05). The content of α2 antigen epitope binding protein in duodenum and jejunum anterior segment of the control group was significantly higher than that in jejunum middle segment,jejunum posterior segment and ileum( P < 0. 05),the content of α2 antigen epitope binding protein in duodenum of the sensitization group was significantly higher than that in and jejunum anterior segment,jejunum middle segment,jejunum posterior segment and ileum( P<0.05). 3) There was a significant difference on content of β4 antigen epitope binding protein in duodenum and jejunum posterior segment between control group and sensitization group( P<0.05). The content of β4 antigen epitope binding protein in duodenum and jejunum anterior segment and jejunum middle segment of control group and sensitization group was significantly higher than that jejunum posterior segment and ileum( P < 0. 05). In conclusion,the content of β-conglycinin,α2 and β4 antigen epitope binding protein in duodenum is the most,and that in jejunum posterior segment and ileum is less. The content of β-conglycinin,α2 and β4 antigen epitope binding protein of sensitization group is generally lower than that of the control group.[Chinese Journal of Animal Nutrition,2019,31(7):3235-3241]
引文
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[7] TALIERCIO E,LOVELESS T M,TURANO MJ,et al.Identification of epitopes of theβsubunit of soybeanβ-conglycinin that are antigenic in pigs,dogs,rabbits and fish[J].Journal of the Science of Food and Agriculture,2014,94(11):2289-2294.
[8] YOU J M,LI D F,QIAO S Y,et al.Development of a monoclonal antibody-based competitive ELISA for detection ofβ-conglycinin,an allergen from soybean[J].Food Chemistry,2008,106(1):352-360.
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[10] ZHAO Y,QIN G X,SUN Z W,et al. Disappearance of immunoreactive glycinin andβ-conglycinin in the digestive tract of piglets[J]. Archives of Animal Nutrition,2008,62(4):322-330.
[11]鲍男.大豆抗原在仔猪小肠组织中分布规律的研究[D].硕士学位论文.长春:吉林农业大学,2007.
[12]马莉.IgA肾病大鼠肠黏膜免疫细胞的变化及大黄酸的调节作用[D].硕士学位论文.南昌:南昌大学,2016.
[13] SUN H,LIU X,WANG Y Z,et al. Allergen-specific immunoglobulin,histamine and T-cell responses induced by soybean glycinin andβ-conglycinin in BALB/c mice of oral sensitisation[J].Food and Agricultural Immunology,2013,24(4):489-501.
[14] LIU X X,LI H R,LU A,et al.Reduction of intestinal mucosal immune function in heat-stressed rats and bacterial translocation[J]. International Journal of Hyperthermia,2012,28(8):756-765.
[2] FU C J,JEZ J M,KERLEY MS,et al.Identification,characterization,epitope mapping,and three-dimensional modeling of theα-subunit ofβ-conglycinin of soybean,a potential allergen for young pigs[J]. Journal of Agricultural and Food Chemistry,2007,55(10):4014-4020.
[3] ZHENG S G,QIN G X,TIAN H,et al. Role of soybeanβ-conglycinin subunits as potential dietary allergens in piglets[J]. The Veterinary Journal,2014,199(3):434-438.
[4] GUO P F,PIAO X S,OU D Y,et al.Characterization of the antigenic specificity of soybean proteinβ-conglycinin and its effects on growth and immune function in rats[J]. Archives of Animal Nutrition,2007,61(3):189-200.
[5]李宝,李玉,马良友,等.大豆抗原蛋白对断奶仔猪细胞因子及肠上皮紧密连接蛋白Claudin-1 mRNA表达的影响[J].中国兽医学报,2015,35(9):1511-1517.
[6]宋青龙,袁翔,张海燕,等.大豆球蛋白的抗营养作用及检测技术研究进展[J].动物营养学报,2017,29(12):4260-4265.
[7] TALIERCIO E,LOVELESS T M,TURANO MJ,et al.Identification of epitopes of theβsubunit of soybeanβ-conglycinin that are antigenic in pigs,dogs,rabbits and fish[J].Journal of the Science of Food and Agriculture,2014,94(11):2289-2294.
[8] YOU J M,LI D F,QIAO S Y,et al.Development of a monoclonal antibody-based competitive ELISA for detection ofβ-conglycinin,an allergen from soybean[J].Food Chemistry,2008,106(1):352-360.
[9]强嘉楠.大豆β-Conglycinin及其主要抗原表位与仔猪肠上皮结合蛋白的分布规律与鉴定的研究[D].硕士学位论文.长春:吉林农业大学,2018.
[10] ZHAO Y,QIN G X,SUN Z W,et al. Disappearance of immunoreactive glycinin andβ-conglycinin in the digestive tract of piglets[J]. Archives of Animal Nutrition,2008,62(4):322-330.
[11]鲍男.大豆抗原在仔猪小肠组织中分布规律的研究[D].硕士学位论文.长春:吉林农业大学,2007.
[12]马莉.IgA肾病大鼠肠黏膜免疫细胞的变化及大黄酸的调节作用[D].硕士学位论文.南昌:南昌大学,2016.
[13] SUN H,LIU X,WANG Y Z,et al. Allergen-specific immunoglobulin,histamine and T-cell responses induced by soybean glycinin andβ-conglycinin in BALB/c mice of oral sensitisation[J].Food and Agricultural Immunology,2013,24(4):489-501.
[14] LIU X X,LI H R,LU A,et al.Reduction of intestinal mucosal immune function in heat-stressed rats and bacterial translocation[J]. International Journal of Hyperthermia,2012,28(8):756-765.