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1. Evaluation of Antioxidant, Anti-Inflammatory and Cytoprotective Properties of Ethanolic Mint Extracts from Algeria on 7-Ketocholesterol-Treated Murine RAW 264.7 Macrophages

Author

Fatiha Brahmi, Thomas Nury, Meryam Debbabi, Samia Hadj-Ahmed, Amira Zarrouk, Michel Prost, Khodir Madani, Lila Boulekbache-Makhlouf, and Gérard Lizard

Subjects
Mentha sp. ethanolic extracts, phenolic compounds, flavonoids, carotenoids, 7-ketocholesterol, antioxidant activity, anti-inflammatory activity, cytoprotection, Therapeutics. Pharmacology, RM1-950
Abstract

The present study consisted in evaluating the antioxidant, anti-inflammatory and cytoprotective properties of ethanolic extracts from three mint species (Mentha spicata L. (MS), Mentha pulegium L. (MP) and Mentha rotundifolia (L.) Huds (MR)) with biochemical methods on murine RAW 264.7 macrophages (a transformed macrophage cell line isolated from ascites of BALB/c mice infected by the Abelson leukemia virus). The total phenolic, flavonoid and carotenoid contents were determined with spectrophotometric methods. The antioxidant activities were quantified with the Kit Radicaux Libres (KRLTM), the ferric reducing antioxidant power (FRAP) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. The MS extract showed the highest total phenolic content, and the highest antioxidant capacity, while the MR extract showed the lowest total phenolic content and the lowest antioxidant capacity. The cytoprotective and anti-inflammatory activities of the extracts were quantified on murine RAW 264.7 macrophages treated with 7-ketocholesterol (7KC; 20 µg/mL: 50 µM) associated or not for 24 h and 48 h with ethanolic mint extracts used at different concentrations (25, 50, 100, 200 and 400 µg/mL). Under treatment with 7KC, an important inhibition of cell growth was revealed with the crystal violet test. This side effect was strongly attenuated in a dose dependent manner with the different ethanolic mint extracts, mainly at 48 h. The most important cytoprotective effect was observed with the MS extract. In addition, the effects of ethanolic mint extracts on cytokine secretion (Interleukin (IL)-6, IL-10, Monocyte Chemoattractant Protein (MCP)-1, Interferon (IFN)-ϒ, Tumor necrosis factor (TNF)-α) were determined at 24 h on lipopolysaccharide (LPS, 0.2 µg/mL)-, 7KC (20 µg/mL)- and (7KC + LPS)-treated RAW 264.7 cells. Complex effects of mint extracts were observed on cytokine secretion. However, comparatively to LPS-treated cells, all the extracts strongly reduce IL-6 secretion and two of them (MP and MR) also decrease MCP-1 and TNF-α secretion. However, no anti-inflammatory effects were observed on 7KC- and (7KC + LPS)-treated cells. Altogether, these data bring new evidences on the potential benefits (especially antioxidant and cytoprotective properties) of Algerian mint on human health.

Published
2018
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2. Protective Effects of α-Tocopherol, γ-Tocopherol and Oleic Acid, Three Compounds of Olive Oils, and No Effect of Trolox, on 7-Ketocholesterol-Induced Mitochondrial and Peroxisomal Dysfunction in Microglial BV-2 Cells

Author

Meryam Debbabi, Thomas Nury, Amira Zarrouk, Nadia Mekahli, Maryem Bezine, Randa Sghaier, Stéphane Grégoire, Lucy Martine, Philippe Durand, Emmanuelle Camus, Anne Vejux, Aymen Jabrane, Lionel Bretillon, Michel Prost, Thibault Moreau, Sofien Ben Ammou, Mohamed Hammami, and Gérard Lizard

Subjects
α-tocopherol, γ-tocopherol, Trolox, oleic acid, mitochondria, peroxisome, murine microglial BV-2 cells, Biology (General), QH301-705.5, Chemistry, QD1-999
Abstract

Lipid peroxidation products, such as 7-ketocholesterol (7KC), may be increased in the body fluids and tissues of patients with neurodegenerative diseases and trigger microglial dysfunction involved in neurodegeneration. It is therefore important to identify synthetic and natural molecules able to impair the toxic effects of 7KC. We determined the impact of 7KC on murine microglial BV-2 cells, especially its ability to trigger mitochondrial and peroxisomal dysfunction, and evaluated the protective effects of α- and γ-tocopherol, Trolox, and oleic acid (OA). Multiple complementary chemical assays, flow cytometric and biochemical methods were used to evaluate the antioxidant and cytoprotective properties of these molecules. According to various complementary assays to estimate antioxidant activity, only α-, and γ-tocopherol, and Trolox had antioxidant properties. However, only α-tocopherol, γ-tocopherol and OA were able to impair 7KC-induced loss of mitochondrial transmembrane potential, which is associated with increased permeability to propidium iodide, an indicator of cell death. In addition, α-and γ-tocopherol, and OA were able to prevent the decrease in Abcd3 protein levels, which allows the measurement of peroxisomal mass, and in mRNA levels of Abcd1 and Abcd2, which encode for two transporters involved in peroxisomal β-oxidation. Thus, 7KC-induced side effects are associated with mitochondrial and peroxisomal dysfunction which can be inversed by natural compounds, thus supporting the hypothesis that the composition of the diet can act on the function of organelles involved in neurodegenerative diseases.

Published
2016
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3. Modulation of Kv3.1b potassium channel level and intracellular potassium concentration in 158N murine oligodendrocytes and BV-2 murine microglial cells treated with 7-ketocholesterol, 24S-hydroxycholesterol or tetracosanoic acid (C24:0)

Author

Rym Ben Khalifa, Amira Zarrouk, Sonia Maatoug, Maryem Bezine, R. Kharrat, Yuqin Wang, Meryam Debbabi, Mohamed El Ayeb, Thomas Nury, Jérôme De Seze, Thibault Moreau, Mohammad Samadi, Anne Vejux, Gérard Lizard, William J. Griffiths, Laboratoire Bio-PeroxIL. Biochimie du peroxysome, inflammation et métabolisme lipidique [Dijon] (BIO-PEROXIL), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Laboratoire des Venins et Biomolécules Thérapeutiques - Laboratory of Venoms and Therapeutic Biomolecules (LR11IPT08), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Université de Monastir - University of Monastir (UM), Swansea University, Laboratoire de Chimie et Physique - Approche Multi-échelle des Milieux Complexes (LCP-A2MC), Université de Lorraine (UL), Hôpital de Hautepierre [Strasbourg], This work was supported by grants from: Univ. Bourgogne (Dijon, France), Univ. El Manar (Tunis, Tunisia), Pasteur Institut (Tunis, Tunisia), ASSAD (Louhans, France) and the Departments of Neurology (Prof. Thibault Moreau, University Hospital, Dijon, France, and Prof. Jérôme de Sèze, University Hospital, Strasbourg, France).

Subjects
Male, 0301 basic medicine, medicine.disease_cause, Biochemistry, Mice, Tetracosanoic acid (C24:0), 24S-hydroxycholesterol, 03 medical and health sciences, 0302 clinical medicine, medicine, Animals, BV-2 cells, Ketocholesterols, Cell Line, Transformed, chemistry.chemical_classification, Reactive oxygen species, Chemistry, Fatty Acids, Neurodegeneration, Kv3.1b, Inflammasome, Oxysterols, General Medicine, medicine.disease, 7-Ketocholesterol, Hydroxycholesterols, 158N cells, Potassium channel, Mitochondria, Cell biology, Oligodendroglia, 030104 developmental biology, Shaw Potassium Channels, Apoptosis, Potassium, Microglia, Reactive Oxygen Species, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, 030217 neurology & neurosurgery, Oxidative stress, Homeostasis, Intracellular, medicine.drug
Abstract

International audience; Little is known about K+ regulation playing major roles in the propagation of nerve impulses, as well as in apoptosis and inflammasome activation involved in neurodegeneration. As increased levels of 7-ketocholesterol (7KC), 24S-hydroxycholesterol (24S-OHC) and tetracosanoic acid (C24:0) have been observed in patients with neurodegenerative diseases, we studied the effect of 24 and/or 48 h of treatment with 7KC, 24S-OHC and C24:0 on Kv3.1b potassium channel level, intracellular K+ concentration, oxidative stress, mitochondrial dysfunction, and plasma membrane permeability in 158N oligodendrocytes and BV-2 microglial cells. In 158N cells, whereas increased level of Kv3.1b was only observed with 7KC and 24S-OHC but not with C24:0 at 24 h, an intracellular accumulation of K+ was always detected. In BV-2 cells treated with 7KC, 24S-OHC and C24:0, Kv3.1b level was only increased at 48 h; intracellular K+ accumulation was found at 24 h with 7KC, 24S-OHC and C24:0, and only with C24:0 at 48 h. Positive correlations between Kv3.1b level and intracellular K+ concentration were observed in 158N cells in the presence of 7KC and 24S-OHC, and in 7KC-treated BV-2 cells at 48 h. Positive correlations were also found between Kv3.1b or the intracellular K+ concentration, overproduction of reactive oxygen species, loss of transmembrane mitochondrial potential and increased plasma membrane permeability in 158N and BV-2 cells. Our data support that the lipid environment affects Kv3.1b channel expression and/or functionality, and that the subsequent rupture of K+ homeostasis is relied with oligodendrocytes and microglial cells damages.

Published
2018

4. Argan Oil-Mediated Attenuation of Organelle Dysfunction, Oxidative Stress and Cell Death Induced by 7-Ketocholesterol in Murine Oligodendrocytes 158N

Author

El Mostafa Karym, Meryam Debbabi, Michel Prost, Maryem Bezine, Randa Sghaier, Anne Vejux, Gérard Lizard, Stéphane Grégoire, Wiem Meddeb, Philippe Durand, Lucy Martine, Amira Zarrouk, Lionel Bretillon, Boubker Nasser, Thomas Nury, Asmaa Badreddine, Thibault Moreau, Mustapha Cherkaoui-Malki, Emmanuelle Prost-Camus, Laboratoire Bio-PeroxIL. Biochimie du peroxysome, inflammation et métabolisme lipidique [Dijon] (BIO-PEROXIL), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Université Hassan 1er [Settat], Université de Monastir - University of Monastir (UM), Faculté de médecine de Sousse [Ibn EL Jazzar], Institut préparatoire aux études scientifiques et techniques [La Marsa] (IPEST), Université de Sousse, Laboratoire des Venins et Biomolécules Thérapeutiques - Laboratory of Venoms and Therapeutic Biomolecules (LR11IPT08), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Centre National de la Recherche Scientifique (CNRS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB), Laboratoire de Recherche Appliquée Spiral [Bourgogne] (LARA SPIRAL), Service de Neurologie générale, vasculaire et dégénérative (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), This work was supported by grants from: University of Bourgogne (Dijon, France), University of Monastir (Monastir, Tunisia), Aides et Services à Domicile (ASSAD) (Louhans, France) and the Association Bourguignonne pour les Aplications en Sciences de l’Information en Médecine (ABASIM, Dijon, France). This work was supported by the Action Intégrée of the Comité Mixte Inter-Universitaire Franco-Marocain (CMIFM, MA/14/310) from the Projet Hubert Curien (PHC) Volubilis/Toubkal program, the Ministère de l’Enseignement Supérieur and the Centre National Pour la Recherche Scientifique et Technique (CNRST, Morocco), the projet PPR from the (CNRST, Morocco), the Ministère des Affaires Etrangères, the Conseil Régional de Bourgogne, and the University of Bourgogne. We also thank Philip Bastable for English corrections (University Hospital, Dijon, France)., Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Laboratoire Bio-PeroxIL. Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique ( Bio-PeroxIL ), Université de Bourgogne ( UB ), Laboratoire Bio-PeroxIL. Biochimie du peroxysome, inflammation et métabolisme lipidique [Dijon] ( BIO-PEROXIL ), Université de Bourgogne ( UB ) -Université Bourgogne Franche-Comté ( UBFC ), Nutrition, Aliments Fonctionnels et Santé Vasculaire, Université de Monastir ( UM ), Centre des Sciences du Goût et de l'Alimentation [Dijon] ( CSGA ), Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Recherche Appliquée Spiral [Bourgogne] ( LARA SPIRAL ), Modélisation mathématique et numérique ( M2N ), Conservatoire National des Arts et Métiers [CNAM] : EA7340, UMR 6578 : Adaptabilité Biologique et Culturelle ( UAABC ), Université de la Méditerranée - Aix-Marseille 2-Centre National de la Recherche Scientifique ( CNRS ), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Laboratoire de Biochimie et Neurosciences [Settat, Maroc], Faculté des Sciences et Techniques de Settat-Université Hassan I [Maroc], University of Bourgogne (Dijon, France) University of Monastir (Monastir, Tunisia) Aides et Services a Domicile (ASSAD) (Louhans, France) Association Bourguignonne pour les Aplications en Sciences de l'Information en Medecine (ABASIM, Dijon, France) Action Integree of the Comite Mixte Inter-Universitaire Franco-Marocain (CMIFM) from the Projet Hubert Curien (PHC) Volubilis/Toubkal program MA/14/310 Ministere de l'Enseignement Superieur Centre National Pour la Recherche Scientifique et Technique (CNRST, Morocco) projet PPR from the (CNRST, Morocco) Ministere des Affaires Etrangeres Conseil Regional de Bourgogne University of Bourgogne, and Lizard, Gérard

Subjects
MESH: Plant Oils/pharmacology, Antioxidant, argan oil, extra virgin olive oil, alpha-tocopherol, 7-ketocholesterol, mitochondria, lysosome, peroxisome, oxiapoptophagy, 158N murine, oligodendrocytes, assay interference compounds, polyunsaturated fatty-acids, smooth-muscle-cells, blood-brain-barrier, pear seed oil, alzheimers-disease, docosahexaenoic acid, mediterranean diet, multiple-sclerosis, medicine.medical_treatment, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Antioxidants/pharmacology, Apoptosis, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Antioxidants, MESH: Lipid Peroxidation, lcsh:Chemistry, Lipid peroxidation, Mice, MESH: Oligodendroglia/drug effects, MESH: Animals, MESH: Oxidative Stress/drug effects, lcsh:QH301-705.5, Spectroscopy, chemistry.chemical_classification, General Medicine, régime méditerranéen, α-tocopherol, 158N murine oligodendrocytes, 3. Good health, maladie d'alzheimer, mitochondrie, MESH: Lysosomes/drug effects, Article, Inorganic Chemistry, 03 medical and health sciences, food, huile d'olive vierge, acide gras polyinsaturé, Food and Nutrition, sclérose en plaques, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, tocopherol, huile d'argan, MESH: Cell Line, Tyrosol, MESH: Ketocholesterols/toxicity, 030104 developmental biology, chemistry, MESH: Mitochondria/drug effects, barrière hematoencephalique, Lipid Peroxidation, 0301 basic medicine, Argan oil, medicine.disease_cause, chemistry.chemical_compound, [SDV.IDA]Life Sciences [q-bio]/Food engineering, Ketocholesterols, 2. Zero hunger, MESH: Peroxisomes/drug effects, Computer Science Applications, Oligodendroglia, Neuroprotective Agents, Biochemistry, Docosahexaenoic acid, Alimentation et Nutrition, Polyunsaturated fatty acid, food.ingredient, MESH: Apoptosis/drug effects, Context (language use), Biology, Catalysis, Cell Line, Peroxisomes, medicine, Animals, Plant Oils, Physical and Theoretical Chemistry, MESH: alpha-Tocopherol/pharmacology, MESH: Mice, MESH: Neuroprotective Agents/pharmacology, Organic Chemistry, Oxidative Stress, lcsh:Biology (General), lcsh:QD1-999, Lysosomes, Oxidative stress
Abstract

PMCID: PMC5666899; International audience; Argan oil is widely used in Morocco in traditional medicine. Its ability to treat cardiovascular diseases is well-established. However, nothing is known about its effects on neurodegenerative diseases, which are often associated with increased oxidative stress leading to lipid peroxidation and the formation of 7-ketocholesterol (7KC) resulting from cholesterol auto-oxidation. As 7KC induces oxidative stress, inflammation and cell death, it is important to identify compounds able to impair its harmful effects. These compounds may be either natural or synthetic molecules or mixtures of molecules such as oils. In this context: (i) the lipid profiles of dietary argan oils from Berkane and Agadir (Morocco) in fatty acids, phytosterols, tocopherols and polyphenols were determined by different chromatographic techniques; and (ii) their anti-oxidant and cytoprotective effects in 158N murine oligodendrocytes cultured with 7KC (25-50 mu M; 24 h) without and with argan oil (0.1% v/v) or -tocopherol (400 mu M, positive control) were evaluated with complementary techniques of cellular and molecular biology. Among the unsaturated fatty acids present in argan oils, oleate (C18:1 n-9) and linoleate (C18:1 n-6) were the most abundant; the highest quantities of saturated fatty acids were palmitate (C16:0) and stearate (C18:0). Several phytosterols were found, mainly schottenol and spinasterol (specific to argan oil), cycloartenol, beta-amyrin and citrostadienol alpha- and gamma-tocopherols were also present. Tyrosol and protocatechic acid were the only polyphenols detected. Argan and extra virgin olive oils have many compounds in common, principally oleate and linoleate, and tocopherols. Kit Radicaux Libres (KRL) and ferric reducing antioxidant power (FRAP) tests showed that argan and extra virgin olive oils have anti-oxidant properties. Argan oils were able to attenuate the cytotoxic effects of 7KC on 158N cells: loss of cell adhesion, cell growth inhibition, increased plasma membrane permeability, mitochondrial, peroxisomal and lysosomal dysfunction, and the induction of oxiapoptophagy (OXIdation + APOPTOsis + autoPHAGY). Altogether, our data obtained in 158N oligodendrocytes provide evidence that argan oil is able to counteract the toxic effects of 7KC on nerve cells, thus suggesting that some of its compounds could prevent or mitigate neurodegenerative diseases to the extent that they are able to cross the blood-brain barrier.

Published
2017

5. Protective Effects of α-Tocopherol, γ-Tocopherol and Oleic Acid, Three Compounds of Olive Oils, and No Effect of Trolox, on 7-Ketocholesterol-Induced Mitochondrial and Peroxisomal Dysfunction in Microglial BV-2 Cells

Author

Sofien Ben Ammou, Anne Vejux, Meryam Debbabi, Stéphane Grégoire, Philippe Durand, Nadia Mekahli, Emmanuelle Camus, Lionel Bretillon, Amira Zarrouk, Maryem Bezine, Lucy Martine, Aymen Jabrane, Gérard Lizard, Mohamed Hammami, Thibault Moreau, Randa Sghaier, Michel Prost, Thomas Nury, Lizard, Gérard, Laboratoire Bio-PeroxIL. Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique (Bio-PeroxIL), Université de Bourgogne (UB), Université de Monastir - University of Monastir (UM), Faculté de médecine de Sousse [Ibn EL Jazzar], Université Tunis El Manar (UTM), Laboratoire des Venins et Biomolécules Thérapeutiques - Laboratory of Venoms and Therapeutic Biomolecules (LR11IPT08), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), OEil, nutrition et signalisation cellulaire [CSGA], Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Recherche Appliquée Spiral [Bourgogne] (LARA SPIRAL), Service de Neurologie générale, vasculaire et dégénérative (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Hôpital Universitaire Sahloul (CHU Sahloul), Université de Bourgogne (Dijon, France), Université de Monastir (Monastir, Tunisia), Université El Manar (Tunis, Tunisia), ASSAD (Louhans, France), Nopal Nutra (Dijon, France), and Departments of Neurology (University Hospital, Dijon, France) and (University Hospital, Sousse, Tunisia)., Université de Tunis El Manar (UTM), Centre National de la Recherche Scientifique (CNRS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB), Laboratoire Bio-PeroxIL. Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique ( Bio-PeroxIL ), Université de Bourgogne ( UB ), Nutrition, Aliments Fonctionnels et Santé Vasculaire, Université de Monastir ( UM ), Centre des Sciences du Goût et de l'Alimentation [Dijon] ( CSGA ), Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS ) -Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Recherche Appliquée Spiral [Bourgogne] ( LARA SPIRAL ), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), and University Hospital of Sahloul

Subjects
0301 basic medicine, Antioxidant, Organes des sens, medicine.medical_treatment, Trolox, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, alpha-Tocopherol, MESH: Antioxidants/pharmacology, MESH: Peroxisomes/drug effects, Antioxidants, biochimie métabolique, lcsh:Chemistry, Lipid peroxidation, Mice, chemistry.chemical_compound, peroxisome, MESH: Animals, lcsh:QH301-705.5, Ketocholesterols, Spectroscopy, Membrane Potential, Mitochondrial, α-tocopherol, γ-tocopherol, oleic acid, mitochondria, murine microglial BV-2 cells, peroxydation lipidique, Neurodegeneration, General Medicine, Peroxisome, 3. Good health, Computer Science Applications, MESH: Membrane Potential, Mitochondrial/drug effects, Biochemistry, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Microglia/metabolism, Alimentation et Nutrition, Microglia, MESH: Mitochondria/pathology, MESH: Mitochondria/drug effects, acide oléique, Programmed cell death, MESH: gamma-Tocopherol/pharmacology, Sensory Organs, Médecine humaine et pathologie, Biology, Article, Catalysis, Cell Line, Inorganic Chemistry, 03 medical and health sciences, MESH: Oleic Acid/pharmacology, Peroxisomes, medicine, Animals, Food and Nutrition, Propidium iodide, Physical and Theoretical Chemistry, Olive Oil, Molecular Biology, MESH: Mice, MESH: Peroxisomes/pathology, maladie neurodégénérative, MESH: Microglia/drug effects, gamma-Tocopherol, MESH: Ketocholesterols/pharmacology, Organic Chemistry, Neurosciences, gène mitochondrial, medicine.disease, MESH: Olive Oil/pharmacology, MESH: Cell Line, Oleic acid, 030104 developmental biology, lcsh:Biology (General), lcsh:QD1-999, chemistry, Neurons and Cognition, Human health and pathology, MESH: alpha-Tocopherol/pharmacology
Abstract

Lipid peroxidation products, such as 7-ketocholesterol (7KC), may be increased in the body fluids and tissues of patients with neurodegenerative diseases and trigger microglial dysfunction involved in neurodegeneration. It is therefore important to identify synthetic and natural molecules able to impair the toxic effects of 7KC. We determined the impact of 7KC on murine microglial BV-2 cells, especially its ability to trigger mitochondrial and peroxisomal dysfunction, and evaluated the protective effects of alpha- and gamma-tocopherol, Trolox, and oleic acid (OA). Multiple complementary chemical assays, flow cytometric and biochemical methods were used to evaluate the antioxidant and cytoprotective properties of these molecules. According to various complementary assays to estimate antioxidant activity, only alpha-, and gamma-tocopherol, and Trolox had antioxidant properties. However, only alpha-tocopherol, gamma-tocopherol and OA were able to impair 7KC-induced loss of mitochondrial transmembrane potential, which is associated with increased permeability to propidium iodide, an indicator of cell death. In addition, alpha-and gamma-tocopherol, and OA were able to prevent the decrease in Abcd3 protein levels, which allows the measurement of peroxisomal mass, and in mRNA levels of Abcd1 and Abcd2, which encode for two transporters involved in peroxisomal beta-oxidation. Thus, 7KC-induced side effects are associated with mitochondrial and peroxisomal dysfunction which can be inversed by natural compounds, thus supporting the hypothesis that the composition of the diet can act on the function of organelles involved in neurodegenerative diseases.

Published
2016
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