鼠李糖乳杆菌GR-1缓解大肠杆菌诱导的奶牛子宫内膜上皮细胞炎性反应  

Lactobacillus rhamnosus GR-1 Alleviates Inflammatory Response of Bovine Endometrial Epithelial Cells Challenged with Escherichia coli

作　　者：张亦弛[1] 黄东风[1] 吕艳秋[1] 姚学军[1] 胡鑫[1] 高晓波[1] 王雪 王天坤[1] ZHANG Yi-chi;HUANG Dong-feng;LYU Yan-qiu;YAO Xue-jun;HU Xin;GAO Xiao-bo;WANG Xue;WANG Tian-kun(Beijing Changping District Center of Animal Disease Prevention and Control(Changping District Comprehensive Test Station,Beijing Livestock Innovation Team),Beijing 102200,China;College of Veterinary Medicine,China Agricultural University,Beijing 100193,China)

机构地区：[1]北京市昌平区动物疫病预防控制中心(北京市家畜创新团队昌平区综合试验站),北京昌平102200 [2]中国农业大学动物医学院,北京海淀100193

出　　处：《中国兽医杂志》2023年第8期6-11,共6页Chinese Journal of Veterinary Medicine

摘　　要：为明确鼠李糖乳杆菌GR-1(L.rhamnosus GR-1)在大肠杆菌感染原代奶牛子宫内膜上皮细胞(BEECs)过程中调节炎性反应的机制,本试验将BEECs分为4个组,分别是对照组(加入5 mL培养基)、E.coli攻菌组(加入5 mL浓度为1×10^(5)CFU/mL大肠杆菌)、L.rhamnosus GR-1单独处理组(提前3 h加入5 mL浓度为1×10^(7)CFU/mL L.rhamnosus GR-1)和L.rhamnosus GR-1保护组(加入5 mL浓度为1×10^(7)CFU/mL L.rhamnosus GR-1孵育3 h后去掉上清液,再加入5 mL浓度为1×10^(5)CFU/mL大肠杆菌);采用光学显微镜观察E.coli对BEECs形态的损害情况,实时荧光定量PCR检测相关基因的转录水平,Western blot检测TLR2和NF-κB2的蛋白表达。结果显示,E.coli攻菌6 h后,与E.coli攻菌组相比,L.rhamnosus GR-1保护组BEECs形态保持完整,且TLR2、TLR4、TNF-αmRNA表达量极显著下调(P<0.01或P<0.001),TLR2和NF-κB2的蛋白水平也极显著下降(P<0.001)。结果表明,L.rhamnosus GR-1可能通过TLRs下调BEECs的炎性因子表达,从而缓解E.coli感染造成的奶牛子宫内膜上皮细胞损伤,为奶牛子宫内膜炎的防治提供数据参考。To investigate the mechanism of Lactobacillus rhamnosus GR-1 regulating inflammatory response in bovine endometrium epithelial cells(BEECs)challenged with Escherichia coli(E.coli),this study divided BEECs into four groups:(1)CONT group:cells were challenged with 5 mL culture medium;(2)ECOL group:cells were challenged with 5 mL 1×10^(5)CFU/mL E.coli suspension;(3)LRGR group:cells were challenged with 5 mL 1×10^(7)CFU/mL L.rhamnosus GR-1 suspension;(4)LRGR+ECOL gourp:cells were pretreated with 5 mL 1×10^(7)CFU/mL L.rhamnosus GR-1 for 3 h before challenged with 5 mL 1×10^(5)CFU/mL E.coli.The damages in cell morphology were observed by optical microscope.The expressions of relevant mRNA and protein related to NF-κB pathway were detected by quantitative real-time PCR and Western blot,respectively.The results showed that after challenge with E.coli for 6 h,compared with ECOL group,the morphology of BEECs in LRGR+ECOL group was normal,the mRNA expressions of TLR 2,TLR 4 and TNF-αin BEECs were significantly down-regulated(P<0.01 or P<0.001),and the protein expressions of TLR2 and NF-κB2 in BEECs were significantly decreased(P<0.001).Thus,L.rhamnosus GR-1 could down-regulate inflammatory factor expression in BEECs through TLRs,and alleviate damage caused by E.coli.This data provides useful technical details for the prevention and treatment of bovine endometritis.

关 键 词：鼠李糖乳杆菌GR-1 大肠杆菌 奶牛子宫内膜上皮细胞 TLR NF-κB2 

分 类 号：S858.23[农业科学—临床兽医学]