食品科学 ›› 2024, Vol. 45 ›› Issue (17): 9-16.doi: 10.7506/spkx1002-6630-20230719-204

• 基础研究 • 上一篇    下一篇

红毛藻多糖抑制大肠杆菌黏附Caco-2细胞的机制

段舒舒, 郑明静, 洪涛, 朱艳冰, 倪辉, 姜泽东   

  1. (1.集美大学海洋食品与生物工程学院,福建 厦门 361021;2.福建省食品微生物与酶工程重点实验室,福建 厦门 361021;3.厦门市食品生物工程技术研究中心,福建 厦门 361021)
  • 出版日期:2024-09-15 发布日期:2024-09-09
  • 基金资助:
    国家自然科学基金面上项目(31972081);福建省科技计划引导性项目(2023N0013)

Inhibitory Mechanism of Escherichia coli Adhesion to Caco-2 Monolayers by Polysaccharides from Bangia fusco-purpurea

DUAN Shushu, ZHENG Mingjing, HONG Tao, ZHU Yanbing, NI Hui, JIANG Zedong   

  1. (1. College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China; 2. Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; 3. Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China)
  • Online:2024-09-15 Published:2024-09-09

摘要: 基于体外人结肠腺癌细胞(Caco-2)单层模型,研究红毛藻多糖(Bangia fusco-purpurea polysaccharide,BFP)对大肠杆菌(Escherichia coli)黏附Caco-2细胞的影响以及潜在作用机制。采用CFDA-SE荧光标记技术分析BFP对E. coli黏附Caco-2细胞单层的影响;利用实时聚合酶链式反应分析BFP对Caco-2细胞表面整合素β1和E. coli黏附素fimH及E. coli黏附Caco-2细胞导致细胞产生的炎症因子(白细胞介素(interleukin,IL)-1β、IL-8、肿瘤坏死因子(tumor necrosis factor,TNF)-α)和细胞紧密连接蛋白(闭锁小带蛋白-1(zonula occludens-1,ZO-1)与Occludin)的基因表达情况,最终基于Western Blot分析验证ZO-1、Occludin蛋白表达。结果表明,BFP在质量浓度为400~800 μg/mL时能够显著抑制E. coli黏附Caco-2细胞单层;BFP主要通过下调Caco-2细胞表面整合素β1和E. coli黏附素基因fimH的表达抑制细菌的黏附。此外,BFP可显著抑制E. coli及其培养上清液诱导Caco-2细胞产生的炎症细胞因子基因表达的上调、紧密连接蛋白(ZO-1与Occludin)及其基因表达的下调。综上所述,BFP能够抑制E. coli对Caco-2细胞单层的黏附,实验结果可为其开发新型的抗菌产品以及BFP的高值利用和精深加工提供参考。

关键词: 红毛藻多糖;大肠杆菌;Caco-2细胞;细菌黏附;肠道稳态

Abstract: In this study, the effect of Bangia fusco-purpurea polysaccharide (BFP) on Escherichia coli adhesion to Caco-2 monolayers and its potential mechanism was studied using an in vitro human colon adenocarcinoma cell (Caco-2) monolayer model. The effect of BFP on the adhesion of E. coli to Caco-2 monolayers was analyzed by fluorescence labeling with carboxyfluorescein diacetate and succinimidyl ester (CFDA-SE), and its effect on the gene expression of integrin β1 in Caco-2 cells, the adhesin FimH in E. coli, and inflammatory factors (IL-1β, IL-8 and TNF-α) induced by E. coli adhesion to Caco-2 cells and the tight junction proteins zonula occludens 1 (ZO-1) and occludin were analyzed using real-time polymerase chain reaction (real-time PCR). Also, the protein expression of ZO-1 and occludin in Caco-2 monolayers was detected using Western Blot analysis. Our results showed that BFP at concentrations of 400–800 μg/mL significantly inhibited the adhesion of E. coli to Caco-2 monolayers mainly by down-regulating the gene expression of integrin β1 in Caco-2 cells and FimH in E. coli. In addition, BFP significantly inhibited the up-regulation of inflammatory cytokine gene expression and the down-regulation of ZO-1 and occludin protein and gene expression induced by in Caco-2 cells induced by E. coli and its culture supernatant. In conclusion, BFP inhibited the adhesion of E. coli to Caco-2 cell monolayers, which will lay a scientific basis for the development of novel antimicrobial products and the high-value utilization and deep processing of BFP.

Key words: Bangia fusco-purpurea polysaccharide; Escherichia coli; Caco-2 cells; bacterial adhesion; intestinal homeostasis

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