重组猪β-防御素1对大肠杆菌的抑制作用

doi:10.11843/j.issn.0366-6964.2017.07.019

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (7): 1342-1348.doi: 10.11843/j.issn.0366-6964.2017.07.019

重组猪β-防御素1对大肠杆菌的抑制作用

郭海勇^1#, 袁洪兴^2#, 王云霄³, 徐瑞涛², 宋勤叶^2*

1. 吉林师范大学生命科学学院, 四平 136000;
2. 河北农业大学动物医学院河北省兽医生物技术工程技术研究中心, 保定 071000;
3. 保定市动物疫病预防控制中心, 保定 071001

收稿日期:2017-02-22 出版日期:2017-07-23 发布日期:2017-07-23
通讯作者: 宋勤叶,教授,博士,E-mail:songqinye@126.com
作者简介:郭海勇(1978-),男,吉林榆树人,副教授,博士,主要从事动物微生物学与免疫学研究,E-mail:guohaiyong78@163.com;袁洪兴(1980-),男,河北邯郸人,硕士,主要从事动物传染病与新型生物制品的研究,E-mail:yuanhongxing@126.com
基金资助:
河北省科技计划项目（13226603D-1<2013>-X）；河北省生猪产业技术体系疫病控制专项资金

Inhibitory Effects of Recombinant Porcine Beta-defensin 1 onEscherichia coli

GUO Hai-yong^1#, YUAN Hong-xing^2#, WANG Yun-xiao³, XU Rui-tao², SONG Qin-ye^2*

1. School of Life Science, Jilin Normal University, Siping 136000, China;
2. Hebei Engineering and Technology Research Center of Veterinary Biotechnology, College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China;
3. The Center for Animal Disease Control and Prevention, Baoding 071001, China

Received:2017-02-22 Online:2017-07-23 Published:2017-07-23

摘要/Abstract

摘要：

旨在评价重组猪β-防御素1（porcine β-defensin 1，pBD1）对大肠杆菌（E.coli）的体外抑制活性及其对大肠杆菌病鸡的治疗效果。应用琼脂板打孔扩散法和微量抑菌试验，分别测定重组pBD1对3株E.coli （O₁₄₁、SJZ1和SJZ2株）的体外抑制效果，继而将40只11 d的雏鸡随机分为5组（n=8），即A（防御素）组、B（抗生素）组、C（防御素与抗生素联合治疗）组、D（感染非治疗）组和E（空白对照）组。A、B、C和D组：同时经颈部皮下和口服途径分别给每只鸡接种1 mL E.coli O₁₄₁株（1.25×10⁹ cfu·mL^-1）；E组：以同样途径分别给每只鸡接种1 mL肉汤。当感染鸡表现临床症状时，A、B、C和D组分别口服pBD1[50 μg·（只·次）^-1]、头孢曲松钠（50 mg·kg^-1，按体重给药，下同）、pBD1和头孢曲松钠[分别为50 μg·（只·次）^-1和50 mg·kg^-1]及等体积的无菌生理盐水，每天1次，连续治疗7 d。结果表明，重组pBD1在体外对3株E.coli均具有明显的抑制活性，抑菌效果随浓度增加而增强。与非治疗组相比，三个治疗组的临床症状和病理变化明显减轻，发病率、死亡率降低，感染后期防御素治疗组、防御素与抗生素联合治疗组的体重极显著高于感染非治疗组（P<0.01）。以上结果显示，重组pBD1对E.coli生长具有明显抑制作用，对大肠杆菌病鸡具有良好的治疗效果；重组pBD1的治疗效果优于头孢曲松钠，有助于病鸡恢复；当pBD1与头孢曲松钠联合使用时，疗效更佳。该研究结果可以为大肠杆菌病的有效治疗提供科学依据，为新型抗菌制剂pBD1的研究与开发奠定重要基础。

Abstract:

The present study aimed to evaluate the antibacterial activity and treatment effect of porcine beta-defensin 1 (pBD1) against E. coli in vitro and in sick chicken infected with E. coli. The inhibitory effects of pBD1 on three E. coli strains (O₁₄₁, SJZ1 and SJZ2) were detected through agar-gel diffusion method and micro-antibacterial assay in vitro, respectively. Then forty 11-day-old chickens were divided into 5 groups (n=8) randomly, including the defensin group (A), antibiotic group (B),defensin and antibiotic group (C), non-treatment group after infection (D) and negative control group (E).Group A, B, C and D were challenged with 1 mL E. coli O₁₄₁(1.25×10⁹ cfu·mL^-1) via the neck subcutaneous injection and oral administration respectively, and Group E was inoculated 1 mL nutrient broth with the same administration routes. While theE. coli -infected chicken showing the clinical symptoms, the group A, B, C and D were given orally pBD1 (50 μg per chicken each time), ceftriaxone sodium (50 mg per kg of body weight), pBD1 (50 μg per chicken each time) and ceftriaxone sodium (50 mg per kg of body weight), and the same volume of sterilized saline water once a day, respectively, and the chickens in each group were continuously treated for seven days. The results showed that the recombinant pBD1 displayed clearly antibacterial activity against E.coli in vitro and this antibacterial effect increased as the concentration of pBD1 rose within certain ranges. Compared with the non-treatment group, the reduction were exhibited in clinical symptoms, pathological changes, morbidity and mortality of the three treated group (A, B and C). The body weights of the group A treated with pBD1 and the group C with defensin and antibiotic were significantly higher (P <0.01) than that of the non-treated group (D) in the later period of infection. These results showed that the recombinant pBD1 has obviously inhibitory effect on E. coli, and has effective therapeutic effects on the sick chicken suffering the colibacillosis. The treatment effect of pBD1 was superior to that of ceftriaxone sodium and helpful to refreshment of the sick chicken. The application of pBD1 combined with ceftriaxone sodium demonstrated the better treatment effect. This study could provide scientific basis for effective treatment of the colibacillosis and lay a solid foundation for the research and development of new type antibacterial agent of pBD1.

中图分类号:

S859.797

郭海勇, 袁洪兴, 王云霄, 徐瑞涛, 宋勤叶. 重组猪β-防御素1对大肠杆菌的抑制作用[J]. 畜牧兽医学报, 2017, 48(7): 1342-1348.

GUO Hai-yong, YUAN Hong-xing, WANG Yun-xiao, XU Rui-tao, SONG Qin-ye. Inhibitory Effects of Recombinant Porcine Beta-defensin 1 onEscherichia coli[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(7): 1342-1348.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2017.07.019

https://www.xmsyxb.com/CN/Y2017/V48/I7/1342

参考文献

[1] GANZ T. The role of antimicrobial peptides in innate immunity[J]. Integr Comp Biol, 2003, 43(2):300-304.
[2] DIAMOND G, ZASLOFF M, ECK H, et al. Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa:peptide isolation and cloning of a cDNA[J]. Proc Natl Acad Sci U S A, 1991, 88(9):3952-3956.
[3] GAO C Y, XU T T, ZHAO Q J, et al. Codon optimization enhances the expression of porcine β-defensin-2 in Escherichia coli[J]. Genet Mol Res, 2015, 14(2):4978-4988.
[4] PRUTHVIRAJ D R, USHA A P, VENKATACHALAPATHY R T. Identification of a novel single nucleotide polymorphism in porcine beta-defensin-1 gene[J]. Asian-Australas J Anim Sci, 2016, 29(3):315-320.
[5] ZHANG G L, WU H, SHI J S, et al. Molecular cloning and tissue expression of porcine β-defensin-1[J]. FEBS Lett, 1998, 424(1-2):37-40.
[6] SHI J S, ZHANG G L, WU H, et al. Porcine epithelial β-defensin 1 is expressed in the dorsal tongue at antimicrobial concentrations[J]. Infect Immun, 1999, 67(6):3121-3127.
[7] LI C L, XU T T, CHEN R B, et al. Cloning, expression and characterization of antimicrobial porcine β defensin 1 in Escherichia coli[J]. Protein Expr Purif, 2013, 88(1):47-53.
[8] SILVA O N, MULDER K C L, BARBOSA A E A D, et al. Exploring the pharmacological potential of promiscuous host-defense peptides:from natural screenings to biotechnological applications[J]. Front Microbiol, 2011, 2:232.
[9] BROWN K L, POON G F T, BIRKENHEAD D, et al. Host defense peptide LL-37 selectively reduces proinflammatory macrophage responses[J]. J Immunol, 2011, 186(9):5497-5505.
[10] CHARNLEY M, MOIR A J G, DOUGLAS C W I, et al. Anti-microbial action of melanocortin peptides and identification of a novel X-Pro-_D/L-Val sequence in Gram-positive and Gram-negative bacteria[J]. Peptides, 2008, 29(6):1004-1009.
[11] 薛现凤, 韩菲菲, 高彦华, 等. 猪β-防御素体外抗菌活性和抗氧化活性研究[J]. 农业生物技术学报, 2012, 20(11):1291-1299.
XUE X F, HAN F F, GAO Y H, et al. In vitro detections of antimicrobial and antioxidant activities of porcine β-defensins[J]. Journal of Agricultural Biotechnology, 2012, 20(11):1291-1299. (in Chinese)
[12] NGUYEN L T, HANEY E F, VOGEL H J. The expanding scope of antimicrobial peptide structures and their modes of action[J]. Trends Biotechnol, 2011, 29(9):464-472.
[13] 袁洪兴, 李红园, 赵驻军, 等. 猪β防御素-1基因克隆及其表达载体的构建[C]//第三届京津冀畜牧兽医科技创新研讨会暨"瑞普杯"新思想、新方法、新观点论坛论文集. 天津:天津市畜牧兽医学会, 北京畜牧兽医学会, 河北省畜牧兽医学会, 2012:267-269.
YUAN H X, LI H Y, ZHAO Z J, et al. Clone of porcine β-defensins-1 gene and construction of its expression vector[C]//The Third Symposium on Animal Husbandry and Veterinary Medicine Scientific and Technological Innovation in Beijing-Tianjin-Hebei Region. Tianjin:Tianjin Institute of Animal Husbandry and Veterinary Medicine, Beijing Institute of Animal Husbandry and Veterinary Medicine, Hebei Provincial Institute of Animal Science and Veterinary Medicine, 2012:267-269. (in Chinese)
[14] HIRSCH T, SPIELMANN M, ZUHAILI B, et al. Human beta-defensin-3 promotes wound healing in infected diabetic wounds[J]. J Gene Med, 2009, 11(3):220-228.
[15] ELAHI S, BUCHANAN R M, ATTAH-POKU S, et al. The host defense peptide beta-defensin 1 confers protection against Bordetella pertussis in newborn piglets[J]. Infect Immun, 2006, 74(4):2338-2352.
[16] VELDHUIZEN E J A, KOOMEN I, ULTEE T, et al. Salmonella serovar specific upregulation of porcine defensins 1 and 2 in a jejunal epithelial cell line[J]. Vet Microbiol, 2009, 136(1-2):69-75.
[17] YANG X, CHENG Y T, TAN M F, et al. Overexpression of porcine beta-defensin 2 enhances resistance to Actinobacillus pleuropneumoniae infection in pigs[J]. Infect Immun, 2015, 83(7):2836-2843.
[18] PENG Z X, WANG A R, XIE L Q, et al. Use of recombinant porcine β-defensin 2 as a medicated feed additive for weaned piglets[J]. Sci Rep, 2016, 6:26790, doi:10.1038/srep26790.
[19] TANG Z R, XU L, SHI B S, et al. Oral administration of synthetic porcine beta-defensin-2 improves growth performance and cecal microbial flora and down-regulates the expression of intestinal toll-like receptor-4 and inflammatory cytokines in weaned piglets challenged with enterotoxigenic Escherichia coli[J]. Anim Sci J, 2016, 87(10):1258-1266.
[20] DONNARUMMA G, PAOLETTI I, FUSCO A, et al. β-defensins:work in progress[M]//DONELLI G. Advances in Microbiology, Infectious Diseases and Public Health. Switzerland:Springer International Publishing, 2016, 901:59-76.
[21] BRUHS A, SCHWARZ T, SCHWARZ A. Prevention and mitigation of experimental autoimmune encephalomyelitis by murine β-defensins via induction of regulatory T cells[J]. J Invest Dermatol, 2016, 136(1):173-181.

重组猪β-防御素1对大肠杆菌的抑制作用

Inhibitory Effects of Recombinant Porcine Beta-defensin 1 onEscherichia coli

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

[1]	王雪莹, 高亢, 蔡吉垚, 张森豪, 解伟纯, 王晓娜, 崔文, 姜艳平, 周晗, 王丽, 乔薪瑗, 徐义刚, 李一经, 唐丽杰. 表达乳铁蛋白肽的猪源罗伊氏乳酸杆菌对断乳仔猪抗霍乱沙门菌感染的效果分析[J]. 畜牧兽医学报, 2021, 52(10): 2874-2886.
[2]	李颖, 冯自立, 白瑜, 张宇航, 任雪怡. 蛋清溶菌酶的改造及其抑菌活性[J]. 畜牧兽医学报, 2021, 52(4): 1094-1102.