Cargando…
Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
C.difficile infection (CDI) is not a merely “gut-confined” disease as toxemia could drive the development of CDI-related extra-intestinal effects. These effects could explain the high CDI-associated mortality, not just justified by diarrhea and dehydration. Here, the extra-intestinal effects of toxi...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760802/ https://www.ncbi.nlm.nih.gov/pubmed/33271969 http://dx.doi.org/10.3390/cells9122575 |
_version_ | 1783627418963017728 |
---|---|
author | Tonon, Federica Di Bella, Stefano Grassi, Gabriele Luzzati, Roberto Ascenzi, Paolo di Masi, Alessandra Zennaro, Cristina |
author_facet | Tonon, Federica Di Bella, Stefano Grassi, Gabriele Luzzati, Roberto Ascenzi, Paolo di Masi, Alessandra Zennaro, Cristina |
author_sort | Tonon, Federica |
collection | PubMed |
description | C.difficile infection (CDI) is not a merely “gut-confined” disease as toxemia could drive the development of CDI-related extra-intestinal effects. These effects could explain the high CDI-associated mortality, not just justified by diarrhea and dehydration. Here, the extra-intestinal effects of toxin A (TcdA) and B (TcdB) produced by C. difficile have been studied in vivo using the zebrafish embryo model. Noteworthy, protective properties of human serum albumin (HSA) towards toxins-induced extra-intestinal effects were also addressed. Zebrafish embryos were treated with TcdA, TcdB and/or HSA at 24 h post-fertilization. Embryos were analyzed for 48 h after treatment to check vital signs and morphological changes. Markers related to cardio-vascular damage and inflammation were evaluated by Real-Time quantitative PCR and/or western blotting. Both toxins induced cardiovascular damage in zebrafish embryos by different mechanisms: (i) direct toxicity (i.e., pericardial edema, cardiac chambers enlargement, endothelial alteration); (ii) increased hormonal production and release (i.e., atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP)), (iii) alteration of the vascular system through the increase of the vascular endothelial growth factor (VEGF-A) levels, as well as of its receptors, (iv) pro-inflammatory response through high cytokines production (i.e., CXCL8, IL1B, IL6 and TNFα) and (v) cell-mediated damage due to the increase in neutrophils number. In addition to cardiovascular damage, we observe skin alteration and inflammation. Finally, our data indicate a protective effect of HSA toward the toxins induced extra-intestinal effects. Together, our findings can serve as a starting point for humans’ studies to substantiate and understand the extra-intestinal effects observed in CDI patients. |
format | Online Article Text |
id | pubmed-7760802 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-77608022020-12-26 Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model Tonon, Federica Di Bella, Stefano Grassi, Gabriele Luzzati, Roberto Ascenzi, Paolo di Masi, Alessandra Zennaro, Cristina Cells Article C.difficile infection (CDI) is not a merely “gut-confined” disease as toxemia could drive the development of CDI-related extra-intestinal effects. These effects could explain the high CDI-associated mortality, not just justified by diarrhea and dehydration. Here, the extra-intestinal effects of toxin A (TcdA) and B (TcdB) produced by C. difficile have been studied in vivo using the zebrafish embryo model. Noteworthy, protective properties of human serum albumin (HSA) towards toxins-induced extra-intestinal effects were also addressed. Zebrafish embryos were treated with TcdA, TcdB and/or HSA at 24 h post-fertilization. Embryos were analyzed for 48 h after treatment to check vital signs and morphological changes. Markers related to cardio-vascular damage and inflammation were evaluated by Real-Time quantitative PCR and/or western blotting. Both toxins induced cardiovascular damage in zebrafish embryos by different mechanisms: (i) direct toxicity (i.e., pericardial edema, cardiac chambers enlargement, endothelial alteration); (ii) increased hormonal production and release (i.e., atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP)), (iii) alteration of the vascular system through the increase of the vascular endothelial growth factor (VEGF-A) levels, as well as of its receptors, (iv) pro-inflammatory response through high cytokines production (i.e., CXCL8, IL1B, IL6 and TNFα) and (v) cell-mediated damage due to the increase in neutrophils number. In addition to cardiovascular damage, we observe skin alteration and inflammation. Finally, our data indicate a protective effect of HSA toward the toxins induced extra-intestinal effects. Together, our findings can serve as a starting point for humans’ studies to substantiate and understand the extra-intestinal effects observed in CDI patients. MDPI 2020-12-01 /pmc/articles/PMC7760802/ /pubmed/33271969 http://dx.doi.org/10.3390/cells9122575 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Tonon, Federica Di Bella, Stefano Grassi, Gabriele Luzzati, Roberto Ascenzi, Paolo di Masi, Alessandra Zennaro, Cristina Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model |
title | Extra-Intestinal Effects of C.
difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model |
title_full | Extra-Intestinal Effects of C.
difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model |
title_fullStr | Extra-Intestinal Effects of C.
difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model |
title_full_unstemmed | Extra-Intestinal Effects of C.
difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model |
title_short | Extra-Intestinal Effects of C.
difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model |
title_sort | extra-intestinal effects of c.
difficile toxin a and b: an in vivo study using the zebrafish embryo model |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760802/ https://www.ncbi.nlm.nih.gov/pubmed/33271969 http://dx.doi.org/10.3390/cells9122575 |
work_keys_str_mv | AT tononfederica extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel AT dibellastefano extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel AT grassigabriele extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel AT luzzatiroberto extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel AT ascenzipaolo extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel AT dimasialessandra extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel AT zennarocristina extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel |