猪源植物乳杆菌调节猪肠上皮细胞对鼠伤寒沙门氏菌感染的抑制  

Primary Study of Lactobacillus plantarum from Porcine on Regulating IPEC-J2 Cell Resistance to Salmonella typhimurium Infection

作　　者：刘芮含 韩永林 石彧萌 孙安琪 汤正旭 胡静涛[1] LIU Ruihan;HAN Yonglin;SHI Yumeng;SUN Anqi;TANG Zhengxu;HU Jingtao(College of Animal Science/College of Veterinary Medicine,Jilin Agricultural University,Jilin Province Animal Microecological Preparation Engineering Research Center,Key Laboratory of Animal Production and Product Quality Safety,Ministry of Education,Changchun 130118,China;Kalaqin Banner Bureau of Agriculture and Animal Husbandry,Kalaqin 024400,China;Jilin Provincial Science Popularization Service Center,Changchun 130021,China)

机构地区：[1]吉林农业大学动物科学技术学院/动物医学院,吉林省动物微生态制剂工程研究中心,动物生产及产品质量安全教育部重点实验室,长春130118 [2]喀喇沁旗农牧局,喀喇沁024400 [3]吉林省科普服务中心,长春130021

出　　处：《经济动物学报》2023年第3期189-195,共7页Journal of Economic Animal

基　　金：吉林省科技发展计划项目(20200201109JC);吉林省大学生创新创业训练项目(202010193026,202010193039)。

摘　　要：为探究猪源植物乳杆菌(Lactobacillus plantarum)对鼠伤寒沙门氏菌(Salmonella typhimurium)的防治效果,以仔猪肠道内容物中筛选到的猪源植物乳杆菌LP13为研究对象,将LP13与猪肠上皮细胞(IPEC-J2cell)分别按照细菌数与细胞数比值(感染复数MOI)为10和100进行体外共培养1 h。通过CCK-8检测LP13对IPEC-J2细胞增殖的影响,流式细胞术检测LP13对IPEC-J2细胞凋亡的影响,利用荧光显微镜和平板计数法评价CFSE标记的LP13对IPEC-J2的黏附效果,并确定LP13与IPEC-J2共培养的最佳比例。添加与LP13相同CFU的鼠伤寒沙门氏菌,通过排斥抑制、置换抑制和竞争抑制试验,进一步明确植物乳杆菌对鼠伤寒沙门氏菌感染的抑制效果。结果显示,LP13对IPEC-J2细胞的增殖与凋亡无显著影响;且在MOI为100时,其对细胞的黏附率更高;在排斥抑制(既预先将LP13于IPEC-J2共培养)时,LP13针对鼠伤寒沙门氏菌对IPEC-J2的黏附和侵袭有一定的抑制作用。试验为植物乳杆菌调节猪肠上皮细胞,预防沙门氏菌感染的研究奠定基础。To investigate the effect of Lactobacillus plantarum from porcine against Salmonella typhimurium,LP13 was screened from the intestinal contents of piglets,and LP13 and porcine intestinal epithelial cells(IPEC-J2) were incubated in vitro at a bacterial:cell ratio(MOI) of 10 and 100 for co-culture 1 h,respectively.The effect of LP13 on the proliferation of IPEC-J2 cells was detected by CCK-8,the effect of LP13 on the apoptosis of IPEC-J2 cells was detected by flow cytometry,the effect of CFSE-labeled LP13 on the adhesion of IPEC-J2 was evaluated by fluorescence microscopy and plate counting,and the optimal action ratio of LP13 co-culture with IPEC-J2 was determined.And then,S.typhimurium with the same CFU as LP13 was added to further clarify the inhibitory effect of L.plantarum on S.typhimurium infection by exclusion inhibition,replacement inhibition and competition inhibition assays.The results showed that LP13 had no significant effect on the proliferation and apoptosis of IPEC-J2 cells;and its adhesion rate to the cells was higher at the LP13 to IPEC-J2 ratio(MOI) of 100;finally,it was found that LP13 showed some inhibition against the adhesion and invasion of S.typhimurium to IPEC-J2 during the rejection inhibition(both pre-culture of LP13 in IPEC-J2).This experiment aims to lay the foundation for the study to reveal that L.p lantarum regulates the prevention of Salmonella infection in porcine intestinal epithelial cells.

关 键 词：植物乳杆菌 鼠伤寒沙门氏菌 猪肠上皮细胞 细胞增殖 细胞凋亡 

分 类 号：S852.61[农业科学—基础兽医学]