胰高血糖素样肽-2对脂多糖应激的IPEC-J2细胞形态和紧密连接相关基因表达的影响  

The Effects of GLP-2 on Cell Morphology and the Gene Expression of Tight Junction in LPS Stressed IPEC-J2 Cells

作　　者：余长松[1] 贾刚[1] 邓秋红[1] 陈小玲[1] 赵华[1] 刘光芒[1] 王康宁[1] 

机构地区：[1]四川农业大学动物营养研究所农业部动物抗病营养与饲料重点实验室,雅安625014

出　　处：《畜牧兽医学报》2015年第4期592-599,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省杰出青年基金(2010JQ0043);教育部博士点基金(2015103110011)

摘　　要：本研究旨在考察胰高血糖素样肽-2(Glucagons-like peptide-2,GLP-2)和脂多糖(Lipopolysaccharide,LPS)对仔猪空肠上皮细胞及其紧密连接(Tight Junctions,TJ)相关蛋白基因表达的影响,并探讨GLP-2调控仔猪肠道TJ蛋白基因表达可能的作用机理。试验一设对照、GLP-2、LPS、LPS+GLP-2共4个组,考查各处理对IPEC-J2细胞形态和紧密连接关键蛋白Occludin、Claudin-1和ZO-1基因表达的影响。结果:添加100nmol·L-1 GLP-2能显著改善IPEC-J2细胞形态,显著提高Occludin、Claudin-1和ZO-1mRNA表达(P<0.01);100μg·mL-1 LPS处理能显著破坏细胞形态、降低IPEC-J2细胞紧密连接蛋白mRNA的表达(P<0.01);在添加LPS基础上添加100nmol·L-1 GLP-2能有效维护细胞形态,显著增加IPEC-J2细胞TJ相关蛋白Occludin、Claudin-1和ZO-1mRNA的表达(P<0.01),增加量分别为46.3%、65.1%和30.3%。试验二考察了添加PI3K特异性抑制剂Wortmannin(Wort)和LY294002(LY)阻断PI3K-Akt-mTOR信号转导途径后Occludin、Claudin-1和ZO-1mRNA表达量的变化,试验共设对照、GLP-2、GLP-2+Wort、GLP-2+LY等4个组。结果:添加抑制剂Wort后可显著降低IPEC-J2细胞Akt、mTOR、Occludin、Claudin-1和ZO-1mRNA的表达(P<0.01),降低量分别为46.9%、50.5%,38.1%、49.6%和18.9%;添加LY后上述mRNA的表达量分别显著降低了67.2%、70.8%,49.6%、60.9%和25.8%(P<0.01)。以上结果表明:添加GLP-2能够有效抑制LPS应激对IPEC-J2细胞形态和TJ相关基因表达的损伤,PI3K-Akt-mTOR信号转导途径可能是GLP-2调控肠道紧密连接蛋白基因表达的重要信号通路之一。The purpose of this research was to investigate the effect of glucagon-like peptide-2(GLP-2)and lipopolysaccharide(LPS)on the gene expression of tight junction in piglets jejunum epithelial IPEC-J2 cells and to deeply discuss the possible mechanism of GLP-2regulating the expression of intestinal TJ gene of the piglet.Trial 1,single factor design was adopted and 4treatments(control,GLP-2,LPS,LPS+GLP-2)were used to test the effect of GLP-2and LPS on the mRNA expression of Occludin,Claudin-1and ZO-1in IPEC-J2 cells.The results showed that:100nmol·L-1 GLP-2could improve cellular morphology and significantly increase the expressionlevel of Occludin,Claudin-1and ZO-1 mRNA in IPEC-J2cell(P<0.01);100μg·mL-1 LPS could significantly destroy cellular morphology and significantly reduce the mRNA expression of TJ in IPEC-J2cells(P<0.01).LPS with 100nmol·L-1 GLP-2could improve cellular morphology and significantly increase the expression of Occludin,Claudin-1and ZO-1mRNA in IPEC-J2cell(P <0.01)for 46.3%,65.1% and 30.3%,respectively.Trial 2,PI3 K specific inhibitor,Wortmannin(Wort)and LY294002(LY),were added to investigate whether GLP-2 modulates TJ's mRNA expression in IPEC-J2 cells through PI3K-Akt-mTOR signal transduction pathway.Four treatments(control,GLP-2,GLP-2+Wort,GLP-2+LY)were designed.The results showed that:100nmol·L-1 GLP-2with 10nmol·L-1 Wort could significantly decrease the expression of Akt,mTOR,Occludin,Claudin-1and ZO-1 mRNA in IPEC-J2cells(P<0.01)for 46.9%,50.5%,38.1%,49.6% and 18.9%,respectively;GLP-2with LY could significantly decrease the expression of Akt,mTOR,Occludin,Claudin-1and ZO-1mRNA in IPEC-J2cells(P<0.01)for67.2%,70.8%,49.6%,60.9% and 25.8%,respectively.In summary,the results showed that GLP-2can effectively inhibit the damnification of TJ mRNA expression by LPS.GLP-2may modulate TJ's mRNA expression through PI3K-Akt-mTOR signal transduction pathway in IPEC-J2 cells.

关 键 词：胰高血糖素样肽-2 脂多糖 仔猪空肠上细胞IPEC-J2 紧密连接 PI3K-Akt-mTOR 

分 类 号：S828[农业科学—畜牧学]