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  

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

作　　者：尚沁沁[1] 黄琴[1] 徐歆[1] 史艳云[1] 李卫芬[1] 余东游[1] 

机构地区：[1]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

出　　处：《动物营养学报》2014年第1期279-286,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：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)

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。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 extracellu- lar 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, re- jection 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 af- fect 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 I（88 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 cy- tokine 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 respon- ses rather than block the adhesion of Escherichia coli K88 and Salmonella choleraesuis to Caco-2 cells.

关 键 词：纳豆芽孢杆菌 CACO-2细胞 细胞活力 黏附 细胞因子 

分 类 号：S816[农业科学—饲料科学]