Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa

Purpose: Ferroptosis is a type of iron-dependent regulated cell death characterized by increased bioavailability of redox-active iron, loss of GPX4 antioxidant capacity, and oxidation of polyunsaturated fatty acid-containing phospholipids mediated by reactive oxygen species (ROS). The aim of this...

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Main Authors: Pablo Contreras-Mellado, Anita Bravo, Fabiola Zambrano, Raúl Sánchez, Rodrigo Boguen, Jennie Risopatrón, Osvaldo Merino, Pamela Uribe
Format: Article
Language:English
Published: Korean Society for Sexual Medicine and Andrology 2025-07-01
Series:The World Journal of Men's Health
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author Pablo Contreras-Mellado
Anita Bravo
Fabiola Zambrano
Raúl Sánchez
Rodrigo Boguen
Jennie Risopatrón
Osvaldo Merino
Pamela Uribe
author_facet Pablo Contreras-Mellado
Anita Bravo
Fabiola Zambrano
Raúl Sánchez
Rodrigo Boguen
Jennie Risopatrón
Osvaldo Merino
Pamela Uribe
author_sort Pablo Contreras-Mellado
collection DOAJ
description Purpose: Ferroptosis is a type of iron-dependent regulated cell death characterized by increased bioavailability of redox-active iron, loss of GPX4 antioxidant capacity, and oxidation of polyunsaturated fatty acid-containing phospholipids mediated by reactive oxygen species (ROS). The aim of this study was to evaluate the effect of oxidative stress induced by arachidonic acid (AA) on ferroptotic cell death in human spermatozoa. Materials and Methods: Spermatozoa from normozoospermic donors were exposed to AA (5, 25, and 50 μM) for 1 hour at 37 °C, including an untreated control. Oxidative stress was confirmed by evaluation of cytosolic and mitochondrial ROS production, viability, mitochondrial membrane potential (ΔΨm) and motility. Subsequently, molecular markers of ferroptosis including iron content, levels of GPX4, SLC7A11, ACSL4, IREB2 and lipid peroxidation were evaluated. The analyses were carried out using either flow cytometry, a microplate reader or confocal laser microscopy. Results: AA-induced oxidative stress showed increased cytosolic and mitochondrial ROS production accompanied by impaired ΔΨm, viability and motility in human spermatozoa. These results were associated with biochemical and molecular markers related to ferroptotic cell death including an increase in iron content in the form of ferrous (Fe2+) ions, SLC7A11, ACSL4, IREB2, a decrease in the level of GPX4, and an increase in the level of lipid peroxidation compared to the untreated control. Conclusions: This study revealed that AA-induced oxidative stress induces cell death with biochemical characteristics of ferroptosis in human spermatozoa, demonstrating another mechanism of alteration of sperm function induced by oxidative stress and could establish new therapeutic objectives to prevent the decrease in sperm quality mediated by oxidative stress.
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spelling doaj-art-91e97cbf0177402a881d202bedbc034e2025-08-20T03:23:52ZengKorean Society for Sexual Medicine and AndrologyThe World Journal of Men's Health2287-42082287-46902025-07-0143363364610.5534/wjmh.240085Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human SpermatozoaPablo Contreras-Mellado0https://orcid.org/0000-0002-5494-8177Anita Bravo1https://orcid.org/0009-0004-1909-1288Fabiola Zambrano2https://orcid.org/0000-0003-4840-0508Raúl Sánchez3https://orcid.org/0000-0003-4853-4282Rodrigo Boguen4https://orcid.org/0000-0002-0325-743XJennie Risopatrón5https://orcid.org/0000-0002-0645-3719Osvaldo Merino6https://orcid.org/0000-0001-8984-8267Pamela Uribe7https://orcid.org/0000-0001-7358-7541Center of Excellence in Translational Medicine-Scientific and Technological Bioresources Nucleus (CEMT-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, ChileCenter of Excellence in Translational Medicine-Scientific and Technological Bioresources Nucleus (CEMT-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, ChileCenter of Excellence in Translational Medicine-Scientific and Technological Bioresources Nucleus (CEMT-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, ChileCenter of Excellence in Translational Medicine-Scientific and Technological Bioresources Nucleus (CEMT-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, ChileDepartment of Diagnostic Processes and Evaluation, Faculty of Health Sciences, Universidad Católica de Temuco, Temuco, ChileCenter of Excellence of Biotechnology in Reproduction (CEBIOR), Universidad de La Frontera, Temuco, ChileCenter of Excellence of Biotechnology in Reproduction (CEBIOR), Universidad de La Frontera, Temuco, ChileCenter of Excellence in Translational Medicine-Scientific and Technological Bioresources Nucleus (CEMT-BIOREN), Faculty of Medicine, Universidad de La Frontera, Temuco, ChilePurpose: Ferroptosis is a type of iron-dependent regulated cell death characterized by increased bioavailability of redox-active iron, loss of GPX4 antioxidant capacity, and oxidation of polyunsaturated fatty acid-containing phospholipids mediated by reactive oxygen species (ROS). The aim of this study was to evaluate the effect of oxidative stress induced by arachidonic acid (AA) on ferroptotic cell death in human spermatozoa. Materials and Methods: Spermatozoa from normozoospermic donors were exposed to AA (5, 25, and 50 μM) for 1 hour at 37 °C, including an untreated control. Oxidative stress was confirmed by evaluation of cytosolic and mitochondrial ROS production, viability, mitochondrial membrane potential (ΔΨm) and motility. Subsequently, molecular markers of ferroptosis including iron content, levels of GPX4, SLC7A11, ACSL4, IREB2 and lipid peroxidation were evaluated. The analyses were carried out using either flow cytometry, a microplate reader or confocal laser microscopy. Results: AA-induced oxidative stress showed increased cytosolic and mitochondrial ROS production accompanied by impaired ΔΨm, viability and motility in human spermatozoa. These results were associated with biochemical and molecular markers related to ferroptotic cell death including an increase in iron content in the form of ferrous (Fe2+) ions, SLC7A11, ACSL4, IREB2, a decrease in the level of GPX4, and an increase in the level of lipid peroxidation compared to the untreated control. Conclusions: This study revealed that AA-induced oxidative stress induces cell death with biochemical characteristics of ferroptosis in human spermatozoa, demonstrating another mechanism of alteration of sperm function induced by oxidative stress and could establish new therapeutic objectives to prevent the decrease in sperm quality mediated by oxidative stress.arachidonic acidferroptosisoxidative stressspermatozoasperm function
spellingShingle Pablo Contreras-Mellado
Anita Bravo
Fabiola Zambrano
Raúl Sánchez
Rodrigo Boguen
Jennie Risopatrón
Osvaldo Merino
Pamela Uribe
Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa
The World Journal of Men's Health
arachidonic acid
ferroptosis
oxidative stress
spermatozoa
sperm function
title Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa
title_full Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa
title_fullStr Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa
title_full_unstemmed Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa
title_short Oxidative Stress Induces Changes in Molecular Markers Associated with Ferroptosis in Human Spermatozoa
title_sort oxidative stress induces changes in molecular markers associated with ferroptosis in human spermatozoa
topic arachidonic acid
ferroptosis
oxidative stress
spermatozoa
sperm function
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