植物乳杆菌对大肠杆菌感染猪肠道上皮细胞形态、存活和免疫应答的影响  

Effects of Lactobacillus plantarum on Morphological,Survival and Immunity Response of Escherichia coli Infected Intestinal Epithelial Cells

作　　者：曾燕霞[1] 王四新[1] 张伟[1] 刘辉[1] 张董燕[1] 王雅民[1] 季海峰[1] 王晶[1] ZENG Yanxia;WANG Sixin;ZHANG Wei;LIU Hui;ZHANG Dongyan;WANG Yamin;JI Haifeng;WANG Jing(Institute of Animal Husbandry and Veterinary Medicine,Beijing 100097,China)

机构地区：[1]北京市农林科学院畜牧兽医研究所,北京100097

出　　处：《动物营养学报》2018年第6期2262-2270,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：北京市农林科学院青年基金(QNJJ201607);北京市农林科学院创新能力建设专项(KJCX20180109);生猪产业技术体系北京市创新团队(BAIC02-2017)

摘　　要：本试验旨在探讨植物乳杆菌对大肠杆菌感染猪肠道上皮细胞(IPEC-J2细胞)形态、存活和免疫应答的影响。在体外条件下,采用吉姆萨和台盼蓝染色方法检测植物乳杆菌对大肠杆菌感染IPEC-J2细胞形态和存活的影响,并采用实时定量荧光PCR方法研究了其对大肠杆菌感染IPEC-J2细胞免疫应答的影响。结果表明:植物乳杆菌能够缓解大肠杆菌引起的IPEC-J2细胞形态损伤。与大肠杆菌处理组相比,植物乳杆菌与大肠杆菌共同处理可显著或极显著降低细胞死亡率(1、2 h,P<0.01;3 h,P<0.05)。植物乳杆菌可显著或极显著抑制大肠杆菌引起的IPEC-J2细胞Toll样模式识别受体2(TLR2)(2、3 h,P<0.01)、Toll样模式识别受体6(TLR6)(2 h,P<0.05;3 h,P<0.01)、NOD样模式识别受体2(NOD2)(1 h,P<0.05;2 h,P<0.01)及炎症因子白细胞介素-6(IL-6)(2、3 h,P<0.01)、白细胞介素-8(IL-8)(2、3 h,P<0.01)mRNA的过表达(P<0.05),还可显著或极显著促进IPEC-J2细胞NOD样模式识别受体1(NOD1)mRNA的表达(2 h,P<0.05;3 h,P<0.01)。结果提示,植物乳杆菌能够缓解大肠杆菌引起的IPEC-J2细胞形态损伤及死亡,并可通过调节模式识别受体TLR2、TLR6和NOD2 mRNA的表达缓解大肠杆菌引起的细胞炎症因子IL-6、IL-8 mRNA的过表达。The aim of this study was to investigate the effects of Lactobacillus plantarum on morphological,survival and immunity response of Escherichia coli infected intestinal epithelial cells(IPEC-J2 cells).Under the conditions of in vitro,the effects of Lactobacillus plantarum on morphological and survival of Escherichia coli infected IPEC-J2 cells were assessed using giemsa staining and trypan blue staining methods,and the effects of Lactobacillus plantarum on immunity response of Escherichia coli infected IPEC-J2 cells were as-sessed using real-time PCR method.The results showed that Lactobacillus plantarum could ameliorate Esche-richia coli-induced disruption of cell morphology on IPEC-J2 cells.Compared with the Escherichia coli treated group,the perecentage of dead cells was significantly decreased by Lactobacillus plantarum and Escherichia coli treated(1,2 h,P<0.01;3 h,P<0.05).The Lactobacillus plantarum could significantly inhibit the mRNA overexpressions of Toll-like receptor 2(TLR2)(2,3 h,P<0.01),Toll-like receptor 6(TLR6)(2 h,P<0.05;3 h,P<0.01),NOD-like receptor 2(NOD2)(1 h,P<0.05;2 h,P<0.01)and inflammatory cyto-kines interleukin-6(IL-6)(2,3 h,P<0.01),interleukin-8(IL-8)(2,3 h,P<0.01)in IPEC-J2 cells in-duced by Escherichia coli,and could promote the NOD-like receptor 1(NOD1)mRNA expression in IPEC-J2 cells(2 h,P<0.05;3 h,P<0.01).The results indicated that Lactobacillus plantarum can ameliorate Escherich-ia coli-induced the disruption of cell morphology and death of IPEC-J2 cells.In addition,it can suppress the Escherichia coli-induced mRNA overexpressions of inflammatory cytokines IL-6 and IL-8 by regulating the pat-tern recognition receptor mRNA expression of TLR2,TLR6 and NOD2.

关 键 词：植物乳杆菌 IPEC.J2细胞 大肠杆菌 损伤 免疫应答 

分 类 号：S828[农业科学—畜牧学] S811.2[农业科学—畜牧兽医]