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Trimethylamine-N-Oxide Instigates NLRP3 Inflammasome Activation and Endothelial Dysfunction
Open accessAbstract
BACKGROUND/AIM: Plasma trimethylamine-N-oxide (TMAO), a product of intestinal microbial metabolism of dietary phosphatidylcholine has been recently associated with atherosclerosis and increased risk of cardiovascular diseases (CVD) in rodents and humans. However, the molecular mechanisms of how TMAO induces atherosclerosis and CVD progression are still unclear. The present study tested whether TMAO induces NLRP3 inflammasome formation and activation and thereby contributes to endothelial injury initiating atherogenesis. METHODS: Inflammasome formation and activation was determined by confocal microscopy, caspase-1 activity was measured by colorimetric assay, IL-1β production was measured using ELISA, cell permeability was determined by microplate reader and ZO-1 expression was determined by western blot analysis and confocal microscopy. In in vivo experiments, TMAO was infused by osmotic pump implantation. RESULTS: TMAO treatment significantly increased the colocalization of NLRP3 with Asc or NLRP3 with caspase-1, caspase-1 activity, IL-1β production, cell permeability in carotid artery endothelial cells (CAECs) compared to control cells. Pretreatment with caspase-1 inhibitor, WEHD or Nlrp3 siRNA abolished the TMAO-induced inflammasome formation, activation and cell permeability in these cells. In addition, we explored the mechanisms by which TMAO activates NLRP3 inflammasomes. TMAO-induced the activation of NLRP3 inflammasomes was associated with both redox regulation and lysosomal dysfunction. In animal experiments, direct infusion of TMAO in mice with partially ligated carotid artery were found to have increased NLRP3 inflammasome formation and IL-1β production in the intima of wild type mice. CONCLUSION: The formation and activation of NLRP3 inflammasomes by TMAO may be an important initiating mechanism to turn on the endothelial inflammatory response leading to endothelial dysfunction.
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In the citing paper
“Another gut-derived metabolite, Trimethylamine N-oxide (TMAO), is primarily produced by the small intestinal microbiota from dietary precursors, including choline, betaine, carnitine, and deoxycarnitine, which are abundant in meat, eggs, and dairy products.”
From The Gut–Brain–Microbiota Connection and Its Role in Autism Spectrum Disorders (2025) · cites this paper as [64]
In the citing paper
“TMAO has been implicated in cardiovascular diseases, contributing to platelet hyperreactivity, increased thrombosis risk, and endothelial dysfunction [63,64,65].”
From The Gut–Brain–Microbiota Connection and Its Role in Autism Spectrum Disorders (2025) · cites this paper as [64]
In the citing paper
“some trials performed in carotid artery endothelial cells (CAEC) revealed that TMAO significantly increases the activation and formation of NLRP3, caspase-1 activity, IL-1β production and cell permeability.”
From Implication of Trimethylamine N-Oxide (TMAO) in Disease: Potential Biomarker or New Therapeutic Target (2018) · cites this paper as [89]
In the citing paper
“Recent investigations have suggested TMAO can impact inflammation via priming and activation of the NLRP3 inflammasome in endothelial cells, as well as the arterial vascular wall in mice, involving mitochondrial reactive oxygen species (ROS) production, thioredoxin interacting protein (TXNIP) and lysosomal destabilization87, 130, 131.”
From Gut microbiota and cardiovascular disease. (2020) · cites this paper as [130]
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Metadata
- Authors
- Boini, K. M., Hussain, T., Li, P.-L., & Koka, S. S.
- Affiliations
- University of Houston (US)
- Virginia Commonwealth University (US)
- Venue
- Cellular Physiology and Biochemistry
- Publisher
- S. Karger AG
- Published
- 2017
- Volume
- 44
- Issue
- 1
- Pages
- 152-162
- ISSN
- 1015-8987 (ISSN-L)
- 1421-9778
- PMC
- PMC5828122
- OpenAlex
- W2767806634
- Semantic Scholar
- de8ac468d4e798c06de304d9dfc8cf3df3a51462
- Cited by
- 302
- References
- 60
- Citations by year
- 2026: 38 · 2025: 38 · 2024: 38 · 2023: 46 · 2022: 38 · 2021: 39 · 2020: 26 · 2019: 25 · 2018: 13 · 2017: 1
- FWCI
- 7.26
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Cellular Physiology and Biochemistry (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institutes of Health
- National Heart, Lung, and Blood Institute (R01HL075316, R01HL057244)
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK104031)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
- Life Sciences › Immunology and Microbiology › Immunology › Atherosclerosis and Cardiovascular Diseases
- Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
- Keywords
- trimethylamine N-oxide, NLRP3 inflammasome, endothelial dysfunction, atherosclerosis, caspase-1, IL-1β, endothelial permeability, redox regulation, lysosomal dysfunction, cardiovascular disease, gut microbiota
- MeSH terms
- NLR Family, Pyrin Domain-Containing 3 Protein, Amino Acid Chloromethyl Ketones, Animals, Carotid Arteries, Cells, Cultured, Male, Methylamines, Mice, Inbred C57BL, Microscopy, Fluorescence, Permeability, Signal Transduction, Caspase 1, RNA Interference, RNA, Small Interfering, Endothelial Cells, Mice, Interleukin-1beta, Inflammasomes, Caspase Inhibitors
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