Interaction between Butyrate and Tumor Necrosis Factor α in Primary Rat Colonocytes
-
Published:2023-01-30
Issue:2
Volume:13
Page:258
-
ISSN:2218-273X
-
Container-title:Biomolecules
-
language:en
-
Short-container-title:Biomolecules
Author:
Souders Christopher L.12, Aristizabal-Henao Juan J.123ORCID, Patuel Sarah J.12, Bowden John A.124, Zubcevic Jasenka15, Martyniuk Christopher J.126ORCID
Affiliation:
1. Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA 2. Center for Environmental and Human Toxicology, University of Florida, Gainesville, FL 32611, USA 3. BERG LLC, 500 Old Connecticut Path, Framingham, MA 01701, USA 4. Department of Chemistry, University of Florida, Gainesville, FL 32611, USA 5. Microbiome Consortium, Center for Hypertension and Precision Medicine, Department of Physiology and Pharmacology, The University of Toledo College of Medicine and Life Sciences, Block Health Science Bldg, 3000 Arlington Ave, Toledo, OH 43614, USA 6. UF Genetics Institute, Interdisciplinary Program in Biomedical Sciences Neuroscience, University of Florida, Gainesville, FL 32611, USA
Abstract
Butyrate, a short-chain fatty acid, is utilized by the gut epithelium as energy and it improves the gut epithelial barrier. More recently, it has been associated with beneficial effects on immune and cardiovascular homeostasis. Conversely, tumor necrosis factor alpha (TNFα) is a pro-inflammatory and pro-hypertensive cytokine. While butyrate and TNFα are both linked with hypertension, studies have not yet addressed their interaction in the colon. Here, we investigated the capacity of butyrate to modulate a host of effects of TNFα in primary rodent colonic cells in vitro. We measured ATP levels, cell viability, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), mitochondrial oxidative phosphorylation, and glycolytic activity in colonocytes following exposure to either butyrate or TNFα, or both. To address the potential mechanisms, transcripts related to oxidative stress, cell fate, and cell metabolism (Pdk1, Pdk2, Pdk4, Spr, Slc16a1, Slc16a3, Ppargc1a, Cs, Lgr5, Casp3, Tnfr2, Bax, Bcl2, Sod1, Sod2, and Cat) were measured, and untargeted liquid chromatography–tandem mass spectrometry (LC-MS/MS) was employed to profile the metabolic responses of colonocytes following exposure to butyrate and TNFα. We found that both butyrate and TNFα lowered cellular ATP levels towards a quiescent cell energy phenotype, characterized by decreased oxygen consumption and extracellular acidification. Co-treatment with butyrate ameliorated TNFα-induced cytotoxicity and the reduction in cell viability. Butyrate also opposed the TNFα-mediated decrease in MMP and mitochondrial-to-intracellular calcium ratios, suggesting that butyrate may protect colonocytes against TNFα-induced cytotoxicity by decreasing mitochondrial calcium flux. The relative expression levels of pyruvate dehydrogenase kinase 4 (Pdk4) were increased via co-treatment of butyrate and TNFα, suggesting the synergistic inhibition of glycolysis. TNFα alone reduced the expression of monocarboxylate transporters slc16a1 and slc16a3, suggesting effects of TNFα on butyrate uptake into colonocytes. Of the 185 metabolites that were detected with LC-MS, the TNFα-induced increase in biopterin produced the only significant change, suggesting an alteration in mitochondrial biogenesis in colonocytes. Considering the reports of elevated colonic TNFα and reduced butyrate metabolism in many conditions, including in hypertension, the present work sheds light on cellular interactions between TNFα and butyrate in colonocytes that may be important in understanding conditions of the colon.
Funder
National Institutes of Health
Subject
Molecular Biology,Biochemistry
Reference105 articles.
1. The Prevalence and Risk Factors of Hypertension among the Urban Population in Southeast Asian Countries: A Systematic Review and Meta-Analysis;Mohammad;Int. J. Hypertens.,2021 2. The global epidemiology of hypertension;Mills;Nat. Rev. Nephrol.,2020 3. Gut microbiota in hypertension;Jose;Curr. Opin. Nephrol. Hypertens.,2015 4. The Gut, Its Microbiome, and Hypertension;Richards;Curr. Hypertens. Rep.,2017 5. Gut Dysbiosis is Linked to Hypertension;Yang;Hypertension,2015
|
|