食淀粉乳杆菌代谢产物对TGEV感染IPEC-J2细胞屏障功能的影响  

Effects of Lactobacillus amylovorus Metabolites on the Barrier Function of IPEC-J2 Cells Infected by TGEV

作　　者：田宗民[1] 张萍[1] 赵毅博[1] 刘文娜[1] 王子敬[1] 张宏宇[1] 张睿[1] 魏萍[1] 

机构地区：[1]东北农业大学,哈尔滨150030

出　　处：《中国畜牧兽医》2015年第8期2194-2203,共10页China Animal Husbandry & Veterinary Medicine

基　　金：黑龙江省教育厅重点项目(12521z001);哈尔滨科技局项目(2014RFXGJ096)

摘　　要：本研究采用荧光定量PCR方法,检测了IPEC-J2细胞不同时间点紧密连接蛋白3(claudin 3,CLDN-3)、细胞角蛋白8(cytokeratin 8,KRT8)、黏蛋白1(mucin 1,MUC1)3种蛋白mRNA的表达量变化。本试验将猪传染性胃肠炎病毒(TGEV)感染IPEC-J2细胞,探索食淀粉乳杆菌代谢产物是否通过维持或改善CLDN-3、KRT8、MUC1蛋白mRNA表达来增强细胞的屏障功能,是否对IPEC-J2细胞感染TGEV后的屏障功能产生影响。试验结果发现,正常细胞对照组MUC1、CLDN-3及KRT8蛋白mRNA表达量随时间没有显著变化(P>0.05),MRS培养基对照组与正常细胞对照组相比没有显著变化(P>0.05),食淀粉乳杆菌代谢产物单独处理组对细胞MUC1、CLDN-3及KRT8蛋白mRNA表达量呈显著上调作用(P<0.05);食淀粉乳杆菌代谢产物+TGEV试验组与病毒对照组相比,处理24、36和48h,MUC1、CLDN-3及KRT8蛋白的表达显著升高(P<0.05)。结果表明,食淀粉乳杆菌代谢产物不仅能提高MUC1、CLDN-3及KRT8在IPEC-J2细胞上的表达,还能负调节TGEV诱导的MUC1、CLDN-3及KRT8在IPEC-J2细胞上的表达,从而加强IPEC-J2细胞对TGEV感染的屏障功能。This study used Real-time PCR to detect the changes of expression quantity of claudin 3 （CLDN-3） ,cytokeratin 8 （KRT8） and mucin 1 （MUC1） three proteins mRNA in IPEC-J2 cells at three different time points. IPEC-J2 cells were infected by TGEV, we explored whether Lactoba- cillus amylovorus metabolites could enhance cell barrier function by maintaining or improving mRNA expression of CLDN-3,KRT8 and MUC1, whether had effects on the barrier function of IPEC-J2 infected by TGEV. The results showed that mRNA expressions of MUC1,CLDN-3 and KRT8 in the normal cell control group did not change significantly with time （P〉0.05） ; MRS medium culture group did not change significantly compared to the normal cell control group（P〉 0.05）; Gene expressions of MUC1 ,CLDN-3 and KRT8 proteins in Lactobacillus amylovorus me- tabolites alone treatment group showed significant time-dependent effect （P〈0.05）. After being treated 24,36 and 48 h,gene expressions of MUC1,CLDN-3 and KRT8 proteins in Lactobacillus amylovorus metabolites ＋ TGEV experiment group was significantly higher than virus control group （P 〈 0. 05）. Lactobacillus amylovorus metabolites could improve MUC1, CLDN-3 and KRT8 expressions in IPEC-J2 cells,while being able to negatively regulate MUtSI,tSLDN-3 and KRT8 expressions that TGEV induced in IPEC-J2 cells,thereby inhencing the barrier function of IPEC-J2 cell against TGEV.

关 键 词：食淀粉乳杆菌 猪传染性胃肠炎病毒 猪小肠上皮细胞 

分 类 号：S852.6[农业科学—基础兽医学]