Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio

Altered body condition and diminished growth in wildlife in the Alberta Oil Sands Region (AOSR) are prompting investigations into the impact of oil sands industrial activity on wildlife in the region. Chemical constituents from bitumen-influenced waters, including oil sands process-affected water (O...

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Main Authors: Laiba Jamshed, Amica Marie-Lucas, Genevieve A. Perono, Gregg T. Tomy, Jim J. Petrik, Richard A. Frank, L. Mark Hewitt, Philippe J. Thomas, Alison C. Holloway
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Journal of Xenobiotics
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Online Access:https://www.mdpi.com/2039-4713/15/3/61
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author Laiba Jamshed
Amica Marie-Lucas
Genevieve A. Perono
Gregg T. Tomy
Jim J. Petrik
Richard A. Frank
L. Mark Hewitt
Philippe J. Thomas
Alison C. Holloway
author_facet Laiba Jamshed
Amica Marie-Lucas
Genevieve A. Perono
Gregg T. Tomy
Jim J. Petrik
Richard A. Frank
L. Mark Hewitt
Philippe J. Thomas
Alison C. Holloway
author_sort Laiba Jamshed
collection DOAJ
description Altered body condition and diminished growth in wildlife in the Alberta Oil Sands Region (AOSR) are prompting investigations into the impact of oil sands industrial activity on wildlife in the region. Chemical constituents from bitumen-influenced waters, including oil sands process-affected water (OSPW), can disrupt endocrine signaling, leading to aberrant lipid accumulation and altered glycemic control in mammals. This study aimed to investigate the effects of naphthenic acid fraction components (NAFCs), derived from OSPW, on energy homeostasis using the McA-RH7777 rat hepatocyte model. Cells were exposed to NAFCs at nominal concentrations of 0, 0.73, 14.7, and 73.4 mg/L for 24 and 48 h. We assessed gene expression related to lipid and glucose metabolism and measured triglyceride accumulation, glucose, and fatty acid uptake. NAFC exposure (14.7 and 73.4 mg/L) reduced triglyceride levels and glucose uptake and increased fatty acid uptake and the expression of beta-oxidation genes, suggesting a metabolic switch from glucose to fatty acid oxidation. This switch in substrate availability signifies a shift in cellular energy dynamics, potentially linked to altered mitochondrial function. To investigate this, we conducted adenosine triphosphate (ATP), mitochondrial membrane potential, and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assays to measure cellular ATP levels, mitochondrial membrane potential, and apoptosis, respectively. At both time points, 73.4 mg/L NAFC exposure resulted in increased ATP levels, induced mitochondrial membrane hyperpolarization, and increased apoptosis. These results suggest that mitochondrial efficiency is compromised, necessitating metabolic adaptations to maintain energy homeostasis. Given that cells exhibit metabolic flexibility that allows them to dynamically respond to changes in substrate availability, we further demonstrated that the kynurenine–tryptophan ratio (KTR) serves as a marker for a shift in energy metabolism under these stress conditions. This work provides a mechanistic framework for understanding how bitumen-derived organic contaminants may disrupt metabolic function in wildlife living in the AOSR. These findings further support the use of molecular markers like KTR to evaluate sub-lethal metabolic stress in environmental health monitoring.
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spelling doaj-art-ebbfd0b458394f76ab3339966ae4b0742025-08-20T02:20:58ZengMDPI AGJournal of Xenobiotics2039-47052039-47132025-04-011536110.3390/jox15030061Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine RatioLaiba Jamshed0Amica Marie-Lucas1Genevieve A. Perono2Gregg T. Tomy3Jim J. Petrik4Richard A. Frank5L. Mark Hewitt6Philippe J. Thomas7Alison C. Holloway8Department of Obstetrics & Gynecology, McMaster University, Hamilton, ON L8S 4L8, CanadaCentre of Oil and Gas Research and Development, University of Manitoba, Winnipeg, MB R3T 2N2, CanadaDepartment of Obstetrics & Gynecology, McMaster University, Hamilton, ON L8S 4L8, CanadaCentre of Oil and Gas Research and Development, University of Manitoba, Winnipeg, MB R3T 2N2, CanadaDepartment of Biomedical Sciences, University of Guelph, Guelph, ON N1G 2W1, CanadaWater Science and Technology Directorate, Environment and Climate Change Canada, Burlington, ON L7S 1A1, CanadaWater Science and Technology Directorate, Environment and Climate Change Canada, Burlington, ON L7S 1A1, CanadaWildlife and Landscape Science Directorate, Environment and Climate Change Canada, National Wildlife Research Center, Ottawa, ON K1A 0H3, CanadaDepartment of Obstetrics & Gynecology, McMaster University, Hamilton, ON L8S 4L8, CanadaAltered body condition and diminished growth in wildlife in the Alberta Oil Sands Region (AOSR) are prompting investigations into the impact of oil sands industrial activity on wildlife in the region. Chemical constituents from bitumen-influenced waters, including oil sands process-affected water (OSPW), can disrupt endocrine signaling, leading to aberrant lipid accumulation and altered glycemic control in mammals. This study aimed to investigate the effects of naphthenic acid fraction components (NAFCs), derived from OSPW, on energy homeostasis using the McA-RH7777 rat hepatocyte model. Cells were exposed to NAFCs at nominal concentrations of 0, 0.73, 14.7, and 73.4 mg/L for 24 and 48 h. We assessed gene expression related to lipid and glucose metabolism and measured triglyceride accumulation, glucose, and fatty acid uptake. NAFC exposure (14.7 and 73.4 mg/L) reduced triglyceride levels and glucose uptake and increased fatty acid uptake and the expression of beta-oxidation genes, suggesting a metabolic switch from glucose to fatty acid oxidation. This switch in substrate availability signifies a shift in cellular energy dynamics, potentially linked to altered mitochondrial function. To investigate this, we conducted adenosine triphosphate (ATP), mitochondrial membrane potential, and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assays to measure cellular ATP levels, mitochondrial membrane potential, and apoptosis, respectively. At both time points, 73.4 mg/L NAFC exposure resulted in increased ATP levels, induced mitochondrial membrane hyperpolarization, and increased apoptosis. These results suggest that mitochondrial efficiency is compromised, necessitating metabolic adaptations to maintain energy homeostasis. Given that cells exhibit metabolic flexibility that allows them to dynamically respond to changes in substrate availability, we further demonstrated that the kynurenine–tryptophan ratio (KTR) serves as a marker for a shift in energy metabolism under these stress conditions. This work provides a mechanistic framework for understanding how bitumen-derived organic contaminants may disrupt metabolic function in wildlife living in the AOSR. These findings further support the use of molecular markers like KTR to evaluate sub-lethal metabolic stress in environmental health monitoring.https://www.mdpi.com/2039-4713/15/3/61AOSRNAFCmitochondrial dysfunctionmetabolic reprogrammingmetabolic switchsubstrate utilization
spellingShingle Laiba Jamshed
Amica Marie-Lucas
Genevieve A. Perono
Gregg T. Tomy
Jim J. Petrik
Richard A. Frank
L. Mark Hewitt
Philippe J. Thomas
Alison C. Holloway
Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio
Journal of Xenobiotics
AOSR
NAFC
mitochondrial dysfunction
metabolic reprogramming
metabolic switch
substrate utilization
title Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio
title_full Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio
title_fullStr Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio
title_full_unstemmed Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio
title_short Naphthenic Acid Fraction Components-Induced Metabolic and Mitochondrial Alterations in Rat Hepatoma Cells: Monitoring Metabolic Reprogramming with Tryptophan–Kynurenine Ratio
title_sort naphthenic acid fraction components induced metabolic and mitochondrial alterations in rat hepatoma cells monitoring metabolic reprogramming with tryptophan kynurenine ratio
topic AOSR
NAFC
mitochondrial dysfunction
metabolic reprogramming
metabolic switch
substrate utilization
url https://www.mdpi.com/2039-4713/15/3/61
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