The anti-adhesive and anti-invasive effects of recombinant azurin on the interaction between enteric pathogens (invasive/non-invasive) and Caco-2 cells

Bakhshi, Bita; Barzelighi, Hajar Mohammadi; Daraei, Bahram

doi:10.1016/j.micpath.2020.104246

Cited by 11 publications

(8 citation statements)

References 43 publications

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“…Bacteria were cultured on Columbia Blood Agar plates and incubated. The single colony was incubated in BHI medium (BHI, Oxoid, U.K.) at 37 °C for 36 h. 28 After that, the bacteria were alternately centrifuged three times in PBS. The pellet was resuspended in DMEM without penicillin and streptomycin.…”

Section: B Vulgatus Intervention On Ulcerative Colitis Micementioning

confidence: 99%

Bergenin Alleviates Ulcerative Colitis By Decreasing Gut Commensal Bacteroides vulgatus-Mediated Elevated Branched-Chain Amino Acids

Huang,

Chen,

Zeng

et al. 2024

J. Agric. Food Chem.

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Ulcerative colitis is closely associated with the dysregulation of gut microbiota. There is growing evidence that natural products may improve ulcerative colitis by regulating the gut microbiota. In this research, we demonstrated that bergenin, a naturally occurring isocoumarin, significantly ameliorates colitis symptoms in dextran sulfate sodium (DSS)-induced mice. Transcriptomic analysis and Caco-2 cell assays revealed that bergenin could ameliorate ulcerative colitis by inhibiting TLR4 and regulating NF-κB and mTOR phosphorylation. 16S rRNA sequencing and metabolomics analyses revealed that bergenin could improve gut microbiota dysbiosis by decreasing branched-chain amino acid (BCAA) levels. BCAA intervention mediated the mTOR/p70S6K signaling pathway to exacerbate the symptoms of ulcerative colitis in mice. Notably, bergenin greatly decreased the symbiotic bacteria Bacteroides vulgatus (B. vulgatus), and the gavage of B. vulgatus increased BCAA concentrations and aggravated the symptoms of ulcerative colitis in mice. Our findings suggest that gut microbiota-mediated BCAA metabolism plays a vital role in the protective effect of bergenin on ulcerative colitis, providing novel insights for ulcerative colitis prevention through manipulation of the gut microbiota.

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Section: B Vulgatus Intervention On Ulcerative Colitis Micementioning

confidence: 99%

Bergenin Alleviates Ulcerative Colitis By Decreasing Gut Commensal Bacteroides vulgatus-Mediated Elevated Branched-Chain Amino Acids

Huang,

Chen,

Zeng

et al. 2024

J. Agric. Food Chem.

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“…When cultured in a 2D interface, Caco-2 cells grow into a confluent monolayer and then differentiate, polarize, and connect to each other, forming an apical brush border structure and expressing genes and proteins relevant to human intestine [ 43 ]. When grown on collagen-coated porous membranes of Transwell inserts, they should represent a valuable model to evaluate transport, barrier and interaction for the intestinal epithelium [ 44 , 45 ]. Another immortalized cell line, HT-29, was isolated from the colorectal adenocarcinoma of a 44-year-old female in 1964 [ 46 ].…”

Section: Current Intestine Modelsmentioning

confidence: 99%

Intestinal Models for Personalized Medicine: from Conventional Models to Microfluidic Primary Intestine-on-a-chip

Chen

Zhao

et al. 2021

Stem Cell Rev and Rep

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Intestinal dysfunction is frequently driven by abnormalities of specific genes, microbiota, or microenvironmental factors, which usually differ across individuals, as do intestinal physiology and pathology. Therefore, it’s necessary to develop personalized therapeutic strategies, which are currently limited by the lack of a simulated intestine model. The mature human intestinal mucosa is covered by a single layer of columnar epithelial cells that are derived from intestinal stem cells (ISCs). The complexity of the organ dramatically increases the difficulty of faithfully mimicking in vivo microenvironments. However, a simulated intestine model will serve as an indispensable foundation for personalized drug screening. In this article, we review the advantages and disadvantages of conventional 2-dimensional models, intestinal organoid models, and current microfluidic intestine-on-a-chip (IOAC) models. The main technological strategies are summarized, and an advanced microfluidic primary IOAC model is proposed for personalized intestinal medicine. In this model, primary ISCs and the microbiome are isolated from individuals and co-cultured in a multi-channel microfluidic chip to establish a microengineered intestine device. The device can faithfully simulate in vivo fluidic flow, peristalsis-like motions, host-microbe crosstalk, and multi-cell type interactions. Moreover, the ISCs can be genetically edited before seeding, and monitoring sensors and post-analysis abilities can also be incorporated into the device to achieve high-throughput and rapid pharmaceutical studies. We also discuss the potential future applications and challenges of the microfluidic platform. The development of cell biology, biomaterials, and tissue engineering will drive the advancement of the simulated intestine, making a significant contribution to personalized medicine in the future. Graphical abstract The intestine is a primary organ for digestion, absorption, and metabolism, as well as a major site for the host-commensal microbiota interaction and mucosal immunity. The complexity of the organ dramatically increases the difficulty of faithfully mimicking in vivo microenvironments, though physiological 3-dimensional of the native small intestinal epithelial tissue has been well documented. An intestinal stem cells-based microfluidic intestine-on-a-chip model that faithfully simulate in vivo fluidic flow, peristalsis-like motions, host-microbe crosstalk, and multi-cell type interactions will make a significant contribution.

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“…Patogenesis S dysenteriae diawali dengan masuknya S dysenteriae ke dalam tubuh inang melalui oral menuju mukosa saluran cerna. Adhesi adalah tahap awal yang sangat penting dalamn, diikuti dengan amplifikasi pada inang, invasi bakteri, cedera jaringan dam penyebaran ke jaringan lain (12)(13)(14)(15).…”

Section: Pendahuluanunclassified

“…Hal ini bisa terjadi karena reseptor permukaan enterosit mencit Balb/c sudah dipenuhi oleh protein pili S. dysenteriae 42 kDa. Dengan demikian adhesi dapat dicegah dan proses pathogenesis tidak berlanjut lagi (15,25) Protein pili S. dysenteriae 42 kDa mampu memediasi terjadinya adhesi, mekanisme ini menunjukkan interaksi antara protein pili S. dysenteriae 42 kDa dengan sel inang yang kuat.…”

Section: Hasilunclassified

Indeks adhesi Shigella dysenteriae pada enterosit mencit galur BALB/ pasca pemaparan protein pili 42 kDa

Suswati¹,

Purnamasari²,

Wisudanti³

et al. 2020

JKS

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Abstrak. Tujuan penelitian ini adalah membuktikan bahwa protein pili Shigella dysenteriae dengan berat molekul 42 kDa merupakan protein adhesi dari S. dysenteriae pada enterosit mencit galur BALB/c. Penelitian ini merupakan penelitian eksperimental laboratoris yang dilaksanakan di Laboratorium Mikrobiologi Fakultas Kedokteran Universitas Jember, melalui beberapa tahap yaitu isolasi dan identifikasi S. dysenteriae, kultur S. dysenteriae, isolasi pili, SDS-PAGE, pemurnian protein pili, uji hemaglutinasi,isolasi enterosit mencit galur BALB/c, dan uji adhesi. Pada penelitian ini dibentuk 6 kelompok perlakuan dan 1 kontrol negatif. Keenam kelompok perlakuan tersebut meliputi konsentrasi protein pili 1, ½, ¼, 1/8, 1/16, dan 1/32. Data dianalisis menggunakan program statistik SPSS (Statistical Product and Service Solution) versi 16, jenis regresi linier sederhana dan one way Anov. Hasil penelitian menunjukkan bahwa protein pili S. dysenteriae 42 kDa mampu menghambat perlekatan S. dysenteriae terhadap enterosit mencit galur BALB/c. Pada uji regresi linier sederhana diperoleh nilai R square 0,897 yang berarti nilai indeks adhesi dipengaruhi oleh konsentrasi protein pili sebesar 89,7%, dan 10,3% dipengaruhi oleh faktor lain. Hasil uji one way Anova menunjukkan bahwa perbedaan konsentrasi protein pili berpengaruh terhadap indeks adhesi bakteri dengan nilai Sig. = 0,000 (p value 0,05). Kesimpulan yang diperoleh dari penelitian ini sesuai dengan hipotesis, yaitu protein pili dengan berat molekul 42 kDa merupakan protein adhesi dari Shigella dysenteriae pada enterosit mencit galur BALB/c. Kata kunci: Shigella dysenteriae, protein pili 42 kDa, indeks adhesi Abstract. The aim of this study was to prove that the protein pili Shigella dysenteriae with a molecular weight of 42 kDa is an adhesion protein from S. dysenteriae in enterocytes of BALB / c mice. This research is a laboratory experimental research carried out at the Microbiology Laboratory of the Faculty of Medicine, University of Jember, through several stages, namely the isolation and identification of S. dysenteriae, culture of S. dysenteriae, isolation of pili, SDS-PAGE, purification of pili protein, hemagglutination test, isolation of enterocyte strain mice. BALB / c, and adhesion test. In this study, 6 treatment groups and 1 negative control were formed. The six treatment groups included protein concentrations of pili 1, ½, ¼, 1/8, 1/16, and 1/32. The data obtained were analyzed using the statistical program SPSS (Statistical Product and Service Solution) version 16, simple linear regression and one way Anov. The results showed that the 42 kDa S. dysenteriae pili protein was able to inhibit the attachment of S. dysenteriae to enterocytes of the BALB / c mice. In the simple linear regression test, it was obtained that the R square value was 0.897, which means that the adhesion index value was influenced by the pili protein concentration of 89.7%, and 10.3% was influenced by other factors. The one way Anova test results showed that the difference in pili protein concentration had an effect on the bacterial adhesion index with the Sig. = 0.000 (p value 0.05). The conclusion obtained from this study is in accordance with the hypothesis, namely pili protein with a molecular weight of 42 kDa is the adhesion protein of Shigella dysenteriae in enterocytes of BALB / c mice. Key words: Shigella dysenteriae, 42 kDa pili protein, adhesion index

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The anti-adhesive and anti-invasive effects of recombinant azurin on the interaction between enteric pathogens (invasive/non-invasive) and Caco-2 cells

Cited by 11 publications

References 43 publications

Bergenin Alleviates Ulcerative Colitis By Decreasing Gut Commensal Bacteroides vulgatus-Mediated Elevated Branched-Chain Amino Acids

Bergenin Alleviates Ulcerative Colitis By Decreasing Gut Commensal Bacteroides vulgatus-Mediated Elevated Branched-Chain Amino Acids

Intestinal Models for Personalized Medicine: from Conventional Models to Microfluidic Primary Intestine-on-a-chip

Indeks adhesi Shigella dysenteriae pada enterosit mencit galur BALB/ pasca pemaparan protein pili 42 kDa

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