红毛藻多糖抑制大肠杆菌黏附Caco-2细胞的机制

TS201.4; 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 fac...

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Published in食品科学 Vol. 45; no. 17; pp. 9 - 16
Main Authors 段舒舒, 郑明静, 洪涛, 朱艳冰, 倪辉, 姜泽东
Format Magazine Article
LanguageChinese
Published 集美大学海洋食品与生物工程学院,福建厦门 361021%集美大学海洋食品与生物工程学院,福建厦门 361021 15.09.2024
厦门市食品生物工程技术研究中心,福建厦门 361021
福建省食品微生物与酶工程重点实验室,福建厦门 361021
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ISSN1002-6630
DOI10.7506/spkx1002-6630-20230719-204

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Abstract TS201.4; 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.
AbstractList TS201.4; 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.
Abstract_FL 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.
Author 郑明静
段舒舒
洪涛
倪辉
姜泽东
朱艳冰
AuthorAffiliation 集美大学海洋食品与生物工程学院,福建厦门 361021%集美大学海洋食品与生物工程学院,福建厦门 361021;福建省食品微生物与酶工程重点实验室,福建厦门 361021;厦门市食品生物工程技术研究中心,福建厦门 361021
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Author_FL HONG Tao
NI Hui
JIANG Zedong
DUAN Shushu
ZHENG Mingjing
ZHU Yanbing
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DocumentTitle_FL Inhibitory Mechanism of Escherichia coli Adhesion to Caco-2 Monolayers by Polysaccharides from Bangia fusco-purpurea
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Issue 17
Keywords Caco-2 cells
肠道稳态
Bangia fusco-purpurea polysaccharide
Escherichia coli
细菌黏附
大肠杆菌
intestinal homeostasis
红毛藻多糖
Caco-2细胞
bacterial adhesion
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Publisher 集美大学海洋食品与生物工程学院,福建厦门 361021%集美大学海洋食品与生物工程学院,福建厦门 361021
厦门市食品生物工程技术研究中心,福建厦门 361021
福建省食品微生物与酶工程重点实验室,福建厦门 361021
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Title 红毛藻多糖抑制大肠杆菌黏附Caco-2细胞的机制
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