Inflammasome-Regulated Pyroptotic Cell Death in Disruption of the Gut-Brain Axis After Stroke

Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after stroke. The purpose of this study is to examine the importance of inflammasome-mediated pyroptosis in disruption of the gut-brain axis after...

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Published in:	Translational stroke research Vol. 13; no. 6; pp. 898 - 912
Main Authors:	Kerr, Nadine A., Sanchez, Juliana, O’Connor, Gregory, Watson, Brant D., Daunert, Sylvia, Bramlett, Helen M., Dietrich, W. Dalton
Format:	Journal Article
Language:	English
Published:	New York Springer US 01.12.2022 Springer Nature B.V
Subjects:	Animals Biomedical and Life Sciences Biomedicine Brain-Gut Axis Cardiology Caspases - metabolism Cell death Colon Cytokines Endothelial Cells - metabolism Fecal incontinence Gut-brain axis Inflammasomes - metabolism Laboratory animals Mice Microbiota Neurology Neurosciences Neurosurgery Original Article Pyroptosis Stroke Thrombotic Stroke Traumatic brain injury Vascular Surgery United States > US Photothrombotic stroke Inflammasome Pyroptosis Gut-brain axis
ISSN:	1868-4483, 1868-601X, 1868-601X
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Abstract	Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after stroke. The purpose of this study is to examine the importance of inflammasome-mediated pyroptosis in disruption of the gut-brain axis after experimental stroke. B6129 mice were subjected to a closed-head photothrombotic stroke. We examined the time course of inflammasome protein expression in brain and intestinal lysate using western blot analysis at 1-, 3-, and 7-days post-injury for caspase-1, interleukin-1β, nod-like receptor protein 3 (NLRP3), and apoptosis speck-like protein containing a caspase-recruiting domain (ASC) and gasdermin-D (GSDMD) cleavage. In a separate group of mice, we processed brain tissue 24 and 72 h after thrombotic stroke for immunohistochemical analysis of neuronal and endothelial cell pyroptosis. We examined intestinal tissue for morphological changes and pyroptosis of macrophages. We performed behavioral tests and assessed gut permeability changes to confirm functional changes after stroke. Our data show that thrombotic stroke induces inflammasome activation in the brain and intestinal tissue up to 7-day post-injury as well as pyroptosis of neurons, cerebral endothelial cells, and intestinal macrophages. We found that thrombotic stroke leads to neurocognitive and motor function deficits as well as increased gut permeability. Finally, the adoptive transfer of serum-derived EVs from stroke mice into naive induced inflammasome activation in intestinal tissues. Taken together, these results provide novel information regarding possible mechanisms underlying gut complications after stroke and the identification of new therapeutic targets for reducing the widespread consequences of ischemic brain injury.
AbstractList	Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after stroke. The purpose of this study is to examine the importance of inflammasome-mediated pyroptosis in disruption of the gut-brain axis after experimental stroke. B6129 mice were subjected to a closed-head photothrombotic stroke. We examined the time course of inflammasome protein expression in brain and intestinal lysate using western blot analysis at 1-, 3-, and 7-days post-injury for caspase-1, interleukin-1β, nod-like receptor protein 3 (NLRP3), and apoptosis speck-like protein containing a caspase-recruiting domain (ASC) and gasdermin-D (GSDMD) cleavage. In a separate group of mice, we processed brain tissue 24 and 72 h after thrombotic stroke for immunohistochemical analysis of neuronal and endothelial cell pyroptosis. We examined intestinal tissue for morphological changes and pyroptosis of macrophages. We performed behavioral tests and assessed gut permeability changes to confirm functional changes after stroke. Our data show that thrombotic stroke induces inflammasome activation in the brain and intestinal tissue up to 7-day post-injury as well as pyroptosis of neurons, cerebral endothelial cells, and intestinal macrophages. We found that thrombotic stroke leads to neurocognitive and motor function deficits as well as increased gut permeability. Finally, the adoptive transfer of serum-derived EVs from stroke mice into naive induced inflammasome activation in intestinal tissues. Taken together, these results provide novel information regarding possible mechanisms underlying gut complications after stroke and the identification of new therapeutic targets for reducing the widespread consequences of ischemic brain injury. Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after stroke. The purpose of this study is to examine the importance of inflammasome-mediated pyroptosis in disruption of the gut-brain axis after experimental stroke. B6129 mice were subjected to a closed-head photothrombotic stroke. We examined the time course of inflammasome protein expression in brain and intestinal lysate using western blot analysis at 1-, 3-, and 7-days post-injury for caspase-1, interleukin-1β, nod-like receptor protein 3 (NLRP3), and apoptosis speck-like protein containing a caspase-recruiting domain (ASC) and gasdermin-D (GSDMD) cleavage. In a separate group of mice, we processed brain tissue 24 and 72 h after thrombotic stroke for immunohistochemical analysis of neuronal and endothelial cell pyroptosis. We examined intestinal tissue for morphological changes and pyroptosis of macrophages. We performed behavioral tests and assessed gut permeability changes to confirm functional changes after stroke. Our data show that thrombotic stroke induces inflammasome activation in the brain and intestinal tissue up to 7-day post-injury as well as pyroptosis of neurons, cerebral endothelial cells, and intestinal macrophages. We found that thrombotic stroke leads to neurocognitive and motor function deficits as well as increased gut permeability. Finally, the adoptive transfer of serum-derived EVs from stroke mice into naive induced inflammasome activation in intestinal tissues. Taken together, these results provide novel information regarding possible mechanisms underlying gut complications after stroke and the identification of new therapeutic targets for reducing the widespread consequences of ischemic brain injury. Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after stroke. The purpose of this study is to examine the importance of inflammasome-mediated pyroptosis in disruption of the gut-brain axis after experimental stroke. B6129 mice were subjected to a closed-head photothrombotic stroke. We examined the time course of inflammasome protein expression in brain and intestinal lysate using western blot analysis at 1-, 3-, and 7-days post-injury for caspase-1, interleukin-1β, nod-like receptor protein 3 (NLRP3), and apoptosis speck-like protein containing a caspase-recruiting domain (ASC) and gasdermin-D (GSDMD) cleavage. In a separate group of mice, we processed brain tissue 24 and 72 h after thrombotic stroke for immunohistochemical analysis of neuronal and endothelial cell pyroptosis. We examined intestinal tissue for morphological changes and pyroptosis of macrophages. We performed behavioral tests and assessed gut permeability changes to confirm functional changes after stroke. Our data show that thrombotic stroke induces inflammasome activation in the brain and intestinal tissue up to 7-day post-injury as well as pyroptosis of neurons, cerebral endothelial cells, and intestinal macrophages. We found that thrombotic stroke leads to neurocognitive and motor function deficits as well as increased gut permeability. Finally, the adoptive transfer of serum-derived EVs from stroke mice into naive induced inflammasome activation in intestinal tissues. Taken together, these results provide novel information regarding possible mechanisms underlying gut complications after stroke and the identification of new therapeutic targets for reducing the widespread consequences of ischemic brain injury.Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after stroke. The purpose of this study is to examine the importance of inflammasome-mediated pyroptosis in disruption of the gut-brain axis after experimental stroke. B6129 mice were subjected to a closed-head photothrombotic stroke. We examined the time course of inflammasome protein expression in brain and intestinal lysate using western blot analysis at 1-, 3-, and 7-days post-injury for caspase-1, interleukin-1β, nod-like receptor protein 3 (NLRP3), and apoptosis speck-like protein containing a caspase-recruiting domain (ASC) and gasdermin-D (GSDMD) cleavage. In a separate group of mice, we processed brain tissue 24 and 72 h after thrombotic stroke for immunohistochemical analysis of neuronal and endothelial cell pyroptosis. We examined intestinal tissue for morphological changes and pyroptosis of macrophages. We performed behavioral tests and assessed gut permeability changes to confirm functional changes after stroke. Our data show that thrombotic stroke induces inflammasome activation in the brain and intestinal tissue up to 7-day post-injury as well as pyroptosis of neurons, cerebral endothelial cells, and intestinal macrophages. We found that thrombotic stroke leads to neurocognitive and motor function deficits as well as increased gut permeability. Finally, the adoptive transfer of serum-derived EVs from stroke mice into naive induced inflammasome activation in intestinal tissues. Taken together, these results provide novel information regarding possible mechanisms underlying gut complications after stroke and the identification of new therapeutic targets for reducing the widespread consequences of ischemic brain injury.
Author	O’Connor, Gregory Sanchez, Juliana Watson, Brant D. Bramlett, Helen M. Kerr, Nadine A. Daunert, Sylvia Dietrich, W. Dalton
Author_xml	– sequence: 1 givenname: Nadine A. surname: Kerr fullname: Kerr, Nadine A. organization: Miami Project to Cure Paralysis, Leonard M. Miller School of Medicine, University of Miami – sequence: 2 givenname: Juliana surname: Sanchez fullname: Sanchez, Juliana organization: Miami Project to Cure Paralysis, Leonard M. Miller School of Medicine, University of Miami – sequence: 3 givenname: Gregory surname: O’Connor fullname: O’Connor, Gregory organization: Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine – sequence: 4 givenname: Brant D. surname: Watson fullname: Watson, Brant D. organization: Peritz Scheinberg Cerebral Vascular Disease Research Laboratory, Department of Neurology, Leonard M. Miller School of Medicine, University of Miami – sequence: 5 givenname: Sylvia surname: Daunert fullname: Daunert, Sylvia organization: Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine – sequence: 6 givenname: Helen M. surname: Bramlett fullname: Bramlett, Helen M. organization: Miami Project to Cure Paralysis, Leonard M. Miller School of Medicine, University of Miami, Department of Neurological Surgery, Leonard M. Miller School of Medicine, University of Miami, Bruce W. Carter Department of Veterans Affairs Medical Center – sequence: 7 givenname: W. Dalton orcidid: 0000-0002-8732-4942 surname: Dietrich fullname: Dietrich, W. Dalton email: ddietrich@miami.edu organization: Miami Project to Cure Paralysis, Leonard M. Miller School of Medicine, University of Miami, Department of Neurological Surgery, Leonard M. Miller School of Medicine, University of Miami
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Keywords	Photothrombotic stroke Inflammasome Pyroptosis Gut-brain axis
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PublicationTitle	Translational stroke research
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Snippet	Approximately 50% of stroke survivors experience gastrointestinal complications. The innate immune response plays a role in changes to the gut-brain axis after...
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SubjectTerms	Animals Biomedical and Life Sciences Biomedicine Brain-Gut Axis Cardiology Caspases - metabolism Cell death Colon Cytokines Endothelial Cells - metabolism Fecal incontinence Gut-brain axis Inflammasomes - metabolism Laboratory animals Mice Microbiota Neurology Neurosciences Neurosurgery Original Article Pyroptosis Stroke Thrombotic Stroke Traumatic brain injury Vascular Surgery
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Title	Inflammasome-Regulated Pyroptotic Cell Death in Disruption of the Gut-Brain Axis After Stroke
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