Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury
Background: Intestinal ischemia and reperfusion (IRI) injury induces acute and long-lasting damage to the neuromuscular compartment and dysmotility. This study aims to evaluate the pathogenetic role of hyaluronan (HA), a glycosaminoglycan component of the extracellular matrix, as a modulator of the...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Language: | English |
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MDPI AG
2022-10-01
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| Online Access: | https://www.mdpi.com/2073-4409/11/21/3370 |
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| author | Annalisa Bosi Davide Banfi Michela Bistoletti Lucia Martina Catizzone Anna Maria Chiaravalli Paola Moretto Elisabetta Moro Evgenia Karousou Manuela Viola Maria Cecilia Giron Francesca Crema Carlo Rossetti Giorgio Binelli Alberto Passi Davide Vigetti Cristina Giaroni Andreina Baj |
| author_facet | Annalisa Bosi Davide Banfi Michela Bistoletti Lucia Martina Catizzone Anna Maria Chiaravalli Paola Moretto Elisabetta Moro Evgenia Karousou Manuela Viola Maria Cecilia Giron Francesca Crema Carlo Rossetti Giorgio Binelli Alberto Passi Davide Vigetti Cristina Giaroni Andreina Baj |
| author_sort | Annalisa Bosi |
| collection | DOAJ |
| description | Background: Intestinal ischemia and reperfusion (IRI) injury induces acute and long-lasting damage to the neuromuscular compartment and dysmotility. This study aims to evaluate the pathogenetic role of hyaluronan (HA), a glycosaminoglycan component of the extracellular matrix, as a modulator of the enteric neuronal and immune function and of the colonic microbiota during in vivo IRI in the rat small intestine. Methods: mesenteric ischemia was induced in anesthetized adult male rats for 60 min, followed by 24 h reperfusion. Injured, sham-operated and non-injured animals were treated with the HA synthesis inhibitor, 4-methylumbelliferone (4-MU 25 mg/kg). Fecal microbiota composition was evaluated by Next Generation Sequencing. Neutrophil infiltration, HA homeostasis and toll like receptor (TLR2 and TLR4) expression in the small intestine were evaluated by immunohistochemical and biomolecular approaches (qRT-PCR and Western blotting). Neuromuscular responses were studied in vitro, in the absence and presence of the selective TLR2/4 inhibitor, Sparstolonin B (SsnB 10, 30 µM). Results: 4-MU significantly reduced IRI-induced enhancement of potentially harmful <i>Escherichia</i> and <i>Enterococcus</i> bacteria. After IRI, HA levels, neutrophil infiltration, and TLR2 and TLR4 expression were significantly enhanced in the <i>muscularis propria,</i> and were significantly reduced to baseline levels by 4-MU. In the injured, but not in the non-injured and sham-operated groups, SsnB reduced both electrical field-stimulated (EFS, 0.1–40 Hz) contractions and EFS-induced (10 Hz) non-cholinergic non-adrenergic relaxations. Conclusions: enhanced HA levels after intestinal IRI favors harmful bacteria overgrowth, increases neutrophil infiltration and promotes the upregulation of bacterial target receptors, TLR2 and TLR4, in the <i>muscularis propria</i>, inducing a pro-inflammatory state. TLR2 and TLR4 activation may, however, underlay a provisional benefit on excitatory and inhibitory neuronal pathways underlying peristalsis. |
| first_indexed | 2024-03-09T19:11:47Z |
| format | Article |
| id | doaj.art-e74c3d4b9ded4278b92e77c95210240b |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T19:11:47Z |
| publishDate | 2022-10-01 |
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| series | Cells |
| spelling | doaj.art-e74c3d4b9ded4278b92e77c95210240b2023-11-24T04:07:34ZengMDPI AGCells2073-44092022-10-011121337010.3390/cells11213370Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion InjuryAnnalisa Bosi0Davide Banfi1Michela Bistoletti2Lucia Martina Catizzone3Anna Maria Chiaravalli4Paola Moretto5Elisabetta Moro6Evgenia Karousou7Manuela Viola8Maria Cecilia Giron9Francesca Crema10Carlo Rossetti11Giorgio Binelli12Alberto Passi13Davide Vigetti14Cristina Giaroni15Andreina Baj16Department of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Pathology, Ospedale di Circolo, ASST-Sette Laghi, 21100 Varese, ItalyDepartment of Pathology, Ospedale di Circolo, ASST-Sette Laghi, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Internal Medicine and Therapeutics, Section of Pharmacology, University of Pavia, 27100 Pavia, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padova, 35131 Padova, ItalyDepartment of Internal Medicine and Therapeutics, Section of Pharmacology, University of Pavia, 27100 Pavia, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Biotechnology and Life Sciences, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyDepartment of Medicine and Surgery, University of Insubria, 21100 Varese, ItalyBackground: Intestinal ischemia and reperfusion (IRI) injury induces acute and long-lasting damage to the neuromuscular compartment and dysmotility. This study aims to evaluate the pathogenetic role of hyaluronan (HA), a glycosaminoglycan component of the extracellular matrix, as a modulator of the enteric neuronal and immune function and of the colonic microbiota during in vivo IRI in the rat small intestine. Methods: mesenteric ischemia was induced in anesthetized adult male rats for 60 min, followed by 24 h reperfusion. Injured, sham-operated and non-injured animals were treated with the HA synthesis inhibitor, 4-methylumbelliferone (4-MU 25 mg/kg). Fecal microbiota composition was evaluated by Next Generation Sequencing. Neutrophil infiltration, HA homeostasis and toll like receptor (TLR2 and TLR4) expression in the small intestine were evaluated by immunohistochemical and biomolecular approaches (qRT-PCR and Western blotting). Neuromuscular responses were studied in vitro, in the absence and presence of the selective TLR2/4 inhibitor, Sparstolonin B (SsnB 10, 30 µM). Results: 4-MU significantly reduced IRI-induced enhancement of potentially harmful <i>Escherichia</i> and <i>Enterococcus</i> bacteria. After IRI, HA levels, neutrophil infiltration, and TLR2 and TLR4 expression were significantly enhanced in the <i>muscularis propria,</i> and were significantly reduced to baseline levels by 4-MU. In the injured, but not in the non-injured and sham-operated groups, SsnB reduced both electrical field-stimulated (EFS, 0.1–40 Hz) contractions and EFS-induced (10 Hz) non-cholinergic non-adrenergic relaxations. Conclusions: enhanced HA levels after intestinal IRI favors harmful bacteria overgrowth, increases neutrophil infiltration and promotes the upregulation of bacterial target receptors, TLR2 and TLR4, in the <i>muscularis propria</i>, inducing a pro-inflammatory state. TLR2 and TLR4 activation may, however, underlay a provisional benefit on excitatory and inhibitory neuronal pathways underlying peristalsis.https://www.mdpi.com/2073-4409/11/21/3370intestinal ischemia/reperfusion injuryenteric nervous systemintestinal neuromuscular functionmicrobiotaTLRs |
| spellingShingle | Annalisa Bosi Davide Banfi Michela Bistoletti Lucia Martina Catizzone Anna Maria Chiaravalli Paola Moretto Elisabetta Moro Evgenia Karousou Manuela Viola Maria Cecilia Giron Francesca Crema Carlo Rossetti Giorgio Binelli Alberto Passi Davide Vigetti Cristina Giaroni Andreina Baj Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury Cells intestinal ischemia/reperfusion injury enteric nervous system intestinal neuromuscular function microbiota TLRs |
| title | Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury |
| title_full | Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury |
| title_fullStr | Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury |
| title_full_unstemmed | Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury |
| title_short | Hyaluronan Regulates Neuronal and Immune Function in the Rat Small Intestine and Colonic Microbiota after Ischemic/Reperfusion Injury |
| title_sort | hyaluronan regulates neuronal and immune function in the rat small intestine and colonic microbiota after ischemic reperfusion injury |
| topic | intestinal ischemia/reperfusion injury enteric nervous system intestinal neuromuscular function microbiota TLRs |
| url | https://www.mdpi.com/2073-4409/11/21/3370 |
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