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益生菌大肠杆菌Nissle 1917抗逆性能、猪肠上皮细胞黏附率及抑菌效果研究

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  • 西南大学动物科技学院, 生物饲料与分子营养室验室, 重庆 400715
李金龙(1992-),男,重庆人,硕士研究生,从事分子营养学与生物饲料资源开发利用研究。E-mail:812453072@qq.com

收稿日期: 2016-10-09

  网络出版日期: 2017-04-14

基金资助

重庆市自然科学基金(cstc2016jcyjA1414);西南大学基本科研业务费(XDJK2015D005);国家973重点基础研究发展计划项目(2013CB127303);国家948项目(2015Z74)

Probiotics Escherichia coli Nissle 1917: Stress Resistance, Swine Intestinal Epithelial Cell Adhesion Rate and Antimicrobial Effects

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  • Key Laboratory for Bio-Feed and Animal Nutrition, College of Animal Science and Technology, Southwest University, Chongqing 400715, China

Received date: 2016-10-09

  Online published: 2017-04-14

摘要

本试验旨在研究益生菌大肠杆菌Nissle 1917(EcN)抗逆性能、猪肠上皮细胞黏附率及抑菌效果。采用体外法对EcN进行生长曲线绘制和耐酸、耐胆盐、耐热性能的测定;以猪肠上皮细胞IPEC-J2细胞为体外细胞模型,考察了EcN对该细胞的黏附率以及对致病菌大肠杆菌K88的黏附抑制率;同时通过蛋白质印迹法检测了EcN对IPEC-J2细胞β-防御素-2和Toll样受体4的水平的影响。结果表明:1)EcN对高酸、高胆盐和高温环境具有一定耐受能力。2)EcN对IPEC-J2细胞的黏附作用以对数期最佳,黏附率达33.96%,显著高于迟缓期、稳定期和衰亡期(P<0.05)。3)EcN对致病菌大肠杆菌K88具有良好的抑制效果,黏附抑制率达87.84%。4)EcN还能上调IPEC-J2细胞β-防御素-2和Toll样受体4水平。结果提示,益生菌EcN具有较好的抗逆性能,能够良好地黏附猪肠上皮细胞,对致病菌大肠杆菌K88具有良好的抑制作用。

本文引用格式

李金龙, 邓欢, 刘金艳, 毛俊霞, 王瑶, 唐志如 . 益生菌大肠杆菌Nissle 1917抗逆性能、猪肠上皮细胞黏附率及抑菌效果研究[J]. 动物营养学报, 2017 , 29(4) : 1241 -1247 . DOI: 10.3969/j.issn.1006-267x.2017.04.020

Abstract

This experiment was conducted to investigate stress resistance, swine intestinal epithelial cell adhesion rate and antimicrobial effects of probiotics Escherichia coli Nissle 1917 (EcN). The growth curve, acid, bile salt and heat tolerance were analyzed using in vitro methods; IPEC-J2 cells of swine intestinal epithelial cells were selected as the in vitro cell model to investigate the effects of EcN on adhesion rate and inhibition rate of adhesion to an pathogenic bacterium Escherichia coli K88; meanwhile, the effects of EcN on levels of β-defensin-2 and toll-like receptor 4 of IPEC-J2 cells were identified by Western blot. The results showed as follows:1) EcN could survive in strong acid, high concentration of bile salt and high temperature. 2) EcN in logarithmic phase had the highest intestinal epithelial cell adhesion effect and its adhesion rate was 33.96%, which was significantly higher than that in retardation phase, stationary phase and decline phase (P<0.05). 3) EcN had good inhibitory effect on pathogenic bacterium Escherichia coli K88 and its inhibition rate of adhesion was 87.84%. 4) EcN could up-regulate the levels of β-defensins-2 and toll-ike receptor 4. In conclusion, the probiotics EcN presents stress resistance, has great swine intestinal epithelial cells adhesion capacity, and prevents attack from pathogenic bacteria Escherichia coli K88.

参考文献

[1] JACOBI C A,MALFERTHEINER P.Escherichia coli Nissle 1917(Mutaflor):new insights into an old probiotic bacterium[J].Digestive Diseases,2011,29(6):600-607.  

[2] VAARALA O.Immunological effects of probiotics with special reference to Lactobacilli[J].Clinical & Experimental Allergy,2003,33(12):1634-1640.  

[3] TREBICHAVSKY I,SPLICHAL I,RADA V,et al.Modulation of natural immunity in the gut by Escherichia coli strain Nissle 1917[J].Nutrition Reviews,2010,68(8):459-464.  

[4] UKENA S N,SINGH A,DRINGENBERG U,et al.Probiotic Escherichia coli Nissle 1917 inhibits leaky gut by enhancing mucosal integrity[J].PLoS One,2007,2(12):e1308.

[5] HERING N A,RICHTER J F,FROMM A,et al.TcpC protein from E. coli Nissle improves epithelial barrier function involving PKCζ and ERK1/2 signaling in HT-29/B6 cells[J].Mucosal Immunology,2014,7(2):369-378.  

[6] SPLICHALOVA A,TREBICHAVSKY I,RADA V,et al.Interference of Bifidobacterium choerinum or Escherichia coli Nissle 1917 with Salmonella Typhimurium in gnotobiotic piglets correlates with cytokine patterns in blood and intestine[J].Clinical & Experiment Immunology,2011,163(2):242-249.  

[7] SCHROEDER B,DUNCKER S,BARTH S,et al.Preventive effects of the probiotic Escherichia coli strain Nissle 1917 on acute secretory diarrhea in a pig model of intestinal infection[J].Digestive Diseases and Sciences,2006,51(4):724-731.  

[8] VALDEBENITO M,CRUMBLISS A L,WINKELMANN G,et al.Environmental factors influence the production of enterobactin,salmochelin,aerobactin,and yersiniabactin in Escherichia coli strain Nissle 1917[J].International Journal of Medical Microbiology,2006,296(8):513-520.  

[9] HATICE B,BELMA A,GULCIN A.The role of resistance to bile salts and acid tolerance of exopolysaccharides (EPSS) produced by yogurt starter bacteria[J].Archives of Biological Sciences,2010,62(2):323-328.  

[10] TURPIN W,HUMBLOT C,GUYOT J P.Genetic screening of functional properties of Lactic acid bacteria in a fermented pearl millet slurry and in the metagenome of fermented starchy foods[J].Applied and Environmental Microbiology,2011,77(24):8722-8734.  

[11] 熊涛,邓耀军,黄涛,等.罗伊氏乳杆菌NCU801对病原菌的粘附抑制性能[J].南昌大学学报,2015,39(2):179-183.

[12] SCHIERACK P,KLETA S,TEDIN K,et al.E. coli Nissle 1917 affects Salmonella adhesion to porcine intestinal epithelial cells[J].PLoS One,2011,6(2):e14712.

[13] SEO E J,WEIBEL S,WEHKAMP J,et al.Construction of recombinant E. coli Nissle 1917(EcN) strains for the expression and secretion of defensins[J].International Journal of Medical Microbiology,2012,302(6):276-287.  

[14] KLETA S,NORDHOFF M,TEDIN K,et al.Role of F1C fimbriae,flagella,and secreted bacterial components in the inhibitory effect of probiotic Escherichia coli Nissle 1917 on atypical enteropathogenic E. coli infection[J].Infection and Immunity,2014,82(5):1801-1812.  

[15] BOUDEAU J,GLASSER A L,JULIEN S,et al.Inhibitory effect of probiotic Escherichia coli strain Nissle 1917 on adhesion to and invasion of intestinal epithelial cells by adherent-invasive E. coli strains isolated from patients with Crohn's disease[J].Alimentary Pharmacology and Therapeutics,2003,18(1):45-56.  

[16] KLETA S,STEINRÜCK H,BREVES G,et al.Detection and distribution of probiotic Escherichia coli Nissle 1917 clones in swine herds in Germany[J].Journal of Applied Microbiology,2006,101(6):1357-1366.  

[17] MALCHOW H A.Crohn's disease and Escherichia coli:a new approach in therapy to maintain remission of colonic Crohn's disease?[J].Journal of Clinical Gastroenterology,1997,25(4):653-658.  

[18] ALTENHOEFER A,OSWALD S,SONNENBORN U,et al.The probiotic Escherichia coli strain Nissle 1917 interferes with invasion of human intestinal epithelial cells by different enteroinvasive bacterial pathogens[J].FEMS Immunology & Medical Microbiology,2004,40(3):223-229.  

[19] HAFEZ M M.Upregulation of intestinal mucin expression by the probiotic bacterium E. coli Nissle 1917[J].Probiotics and Antimicrobial Proteins,2012,4(2):67-77.  
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