研究简报 Short Communications

纳豆芽孢杆菌B4对Caco-2细胞活性、黏附与细胞因子分泌的影响

  • 尚沁沁 ,
  • 黄琴 ,
  • 徐歆 ,
  • 史艳云 ,
  • 李卫芬 ,
  • 余东游
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  • 浙江大学动物科学学院饲料科学研究所, 农业部华东动物营养与饲料重点实验室, 杭州 310058

收稿日期: 2013-07-15

  网络出版日期: 2013-12-24

基金资助

东南沿海地区大学农业科技服务技术集成与示范项目(2013BAD20B00);杭州市科技局农业科研攻关专项(20120232B26);国家863资金资助项目(2013AA102800)

Effects of Bacillus subtilis var. natto B4 on Activity, Adhesion and Cytokine Secretion of Caco-2 Cells

  • SHANG Qinqin ,
  • HUANG Qin ,
  • XU Xin ,
  • SHI Yanyun ,
  • LI Weifen ,
  • YU Dongyou
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  • Key Laboratory of Animal Nutrition and Feed Science in East China, Ministry of Agriculture, Institute of Feed Science, College of Animal Science, Zhejiang University, Hangzhou 310058, China

Received date: 2013-07-15

  Online published: 2013-12-24

摘要

本试验以Caco-2细胞为体外模型研究纳豆芽孢杆菌B4对其活性、黏附与细胞因子分泌的影响。采用台盼蓝排斥试验和乳酸脱氢酶细胞毒性检测研究纳豆芽孢杆菌B4对Caco-2细胞活性的影响;采用荧光标记法评价大肠杆菌K88和霍乱沙门氏菌的黏附性能,并通过竞争、排斥和置换试验检测纳豆芽孢杆菌B4对这2株病原菌黏附的阻断作用;采用酶联免疫试剂盒测定Caco-2细胞分泌的细胞因子含量。结果表明:1)纳豆芽孢杆菌B4对Caco-2细胞存活率无显著影响(P>0.05),且对其无毒害作用。2)纳豆芽孢杆菌B4对大肠杆菌K88和霍乱沙门氏菌黏附Caco-2细胞的黏附率无显著影响(P>0.05),且对这2株病原菌黏附Caco-2细胞无阻断作用。3)纳豆芽孢杆菌B4极显著提高了Caco-2细胞分泌促炎细胞因子增殖诱导配体含量(P<0.01),对白细胞介素-6和白细胞介素-8含量无显著影响(P>0.05),极显著降低了Caco-2细胞分泌抗炎细胞因子白细胞介素-10含量(P<0.01)。综上所述,纳豆芽孢杆菌B4对Caco-2细胞无毒性,对大肠杆菌K88和霍乱沙门氏菌黏附Caco-2细胞无阻断作用,可通过调节Caco-2细胞的细胞因子分泌发挥一定的免疫功能。

本文引用格式

尚沁沁 , 黄琴 , 徐歆 , 史艳云 , 李卫芬 , 余东游 . 纳豆芽孢杆菌B4对Caco-2细胞活性、黏附与细胞因子分泌的影响[J]. 动物营养学报, 2014 , 26(1) : 279 -286 . DOI: 10.3969/j.issn.1006-267x.2014.01.036

Abstract

Using Caco-2 cells as an in vitro model to investigate the effects of Bacillus subtilis var. natto B4 on the activity, adhesion and cytokine secretion of Caco-2 cells. The trypan blue dye exclusion test and extracellular lactate dehydrogenase activity determination were conducted to research the influence of Bacillus subtilis var. natto B4 on the activity of Caco-2 cells; the fluorescence labeling technology was used to investigate the adhesive properties of Escherichia coli K88 and Salmonella choleraesuis to Caco-2 cells. The competition, rejection and displacement experiments were carried out to determine the blocking action of Bacillus subtilis var. natto B4 to the adhesion of the two pathogenic bacterial strains; the cytokines secreted by Caco-2 cells were measured by ELISA. The results showed as follows: 1) Bacillus subtilis var. natto B4 did not significantly affect survival rate of Caco-2 cells (P>0.05) and had no cytotoxicity on Caco-2 cells. 2) Bacillus subtilis var. natto B4 had no significant effects on the adhesion rate of Escherichia coli K88 and Salmonella choleraesuis to Caco-2 cells (P>0.05), and had no blocking action on the adhesion of the two pathogenic bacterial strains to Caco-2 cells. 3) Bacillus subtilis var. natto B4 significantly increased the content of pro-inflammatory cytokine inducing ligand (P<0.01), and had no significant influence on the contents of interleukin-6 and interleukin-8 (P>0.05), whereas Bacillus subtilis var. natto B4 significantly reduced the content of anti-inflammatory cytokine interleukin-10 (P<0.01). In conclusion, Bacillus subtilis var. natto B4 has no cytotoxicity against Caco-2 cells, and can modulate the cytokine secretion function of Caco-2 cells to induce some immune responses rather than block the adhesion of Escherichia coli K88 and Salmonella choleraesuis to Caco-2 cells.

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