Nitro-Oxidative Stress and Mitochondrial Dysfunction in Human Cell Lines Exposed to the Environmental Contaminants PFOA and BPA

Maria Chiara Magnifico; Marla Xhani; Benedetta Sprovera; Brigitta Buttari; Giorgia Abballe; Flaminia Desideri; Emiliano Panieri; Luciano Saso; Marzia Arese

doi:10.31083/j.fbl2710292

Nitro-Oxidative Stress and Mitochondrial Dysfunction in Human Cell Lines Exposed to the Environmental Contaminants PFOA and BPA

Front Biosci (Landmark Ed). 2022 Oct 27;27(10):292. doi: 10.31083/j.fbl2710292.

Authors

Maria Chiara Magnifico^{1

2}, Marla Xhani^{1

3}, Benedetta Sprovera¹, Brigitta Buttari⁴, Giorgia Abballe¹, Flaminia Desideri¹, Emiliano Panieri^{5

6}, Luciano Saso⁵, Marzia Arese¹

Affiliations

¹ Department of Biochemical Sciences "A. Rossi Fanelli", Sapienza University of Rome, 00185 Rome, Italy.
² Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari "Aldo Moro", 70121 Bari, Italy.
³ Istituto Zooprofilattico Sperimentale del Lazio e della Toscana "M. Aleandri", 00178 Rome, Italy.
⁴ Department of Cardiovascular, Endocrine-Metabolic Diseases and Aging, Italian National Institute of Health, 00161 Rome, Italy.
⁵ Department of Physiology and Pharmacology "Vittorio Erspamer", Sapienza University of Rome, 00185 Rome, Italy.
⁶ Section of Hazardous Substances, Environmental Education and Training for the Technical Coordination of Management Activities (DGTEC), Italian Institute for Environmental Protection and Research, 00144 Rome, Italy.

PMID: 36336871
DOI: 10.31083/j.fbl2710292

Abstract

Background: Bisphenol A (BPA) and perfluorooctanoic acid (PFOA) are synthetic compounds widely utilized in industrial activities devoted to the production of daily life plastic, metal products, and packaging from which they are able to migrate to food and water. Due to their persistence in the environment, living organisms are chronically exposed to these pollutants. BPA and PFOA have adverse effects on tissues and organs. The aim of this study was to identify the molecular targets and biochemical mechanisms involved in their toxicity.

Methods: HepG2 and HaCaT cells were treated with BPA or PFOA, and the trypan blue exclusion test and 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay were performed to define the conditions for subsequent investigations. We conducted quantitative PCR and western blot analysis to evaluate the expression of proteins involved in nitric oxide (NO) signaling. Cell-based assays were carried out to evaluate reactive oxygen species (ROS) production, nitrite/nitrate (NOx) accumulation, 3-nitrotyrosine (3-NT) formation, and mitochondrial membrane potential (MMP) determination in treated cells.

Results: HepG2 and HaCaT cells incubated for 24 h with subtoxic concentrations of BPA or PFOA (50 and 10 μM, respectively) exhibited altered mRNA and protein expression levels of NO synthase isoforms, manganese superoxide dismutase, and cytochrome c. Treatment with PFOA led to activation of inducible NO synthase (NOS), a marker of nitrosative stress, accompanied by the increased production of ROS, NOx, and 3-NT and alterations of the MMP compared to controls.

Conclusions: The results of this study indicate the major involvement of the NO signaling axis in the persistent alteration of cell redox homeostasis and mitochondrial dysfunction induced by BPA and PFOA, highlighting the specific role of PFOA in NOS regulation and induction of nitro-oxidative stress.

Keywords: emerging contaminants; endocrine disruptors; mitochondrial dysfunction; nitric oxide signaling; nitro-oxidative stress.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Humans
Mitochondria* / metabolism
Oxidative Stress*
Reactive Oxygen Species / metabolism

Substances

perfluorooctanoic acid
bisphenol A
Reactive Oxygen Species