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Mitochondrial dysfunction,oxidative stress and apoptotic induction in microglial BV-2 cells treated with sodium arsenate

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Journal of Environmental Sciences 2017年第1期29卷 44-51页

作者： Wafa Kharroubi Samia Hai Ahmed thomas nury Pierre Andreoletti Rachid Sakly Mohamed Hammami Gerard Lizard Univ.Bourgogne Franche-Comte Laboratory Bio-PeroxIL Biochemistry of the PeroxisomeInflammation and Lipid Metabolism EA7270/INSERMFaculty of Sciences GabrielDijon 21000France Laboratory of Nutrition-Functional Foods and Vascular Diseases Faculty of MedicineUniversity of MonastirMonastir 5019Tunisia

The treatment of microglial BV-2 cells with sodium arsenate（As（V）：0.1-400 μmol/L — 48 hr）induces a dose-dependent *** neurotoxic effects of high concentrations of As（V）（100,200 and 400 μmol/L） are characterized by increased levels of mitochondrial complexesⅠ,Ⅱ,and Ⅳ followed by increased superoxide anion ***,As（V） triggers an apoptotic mode of cell death,demonstrated by an apoptotic SubG1 peak,associated with an alteration of plasma membrane *** is also a decrease in transmembrane mitochondrial potential and mitochondrial adenosine triphosphate *** is therefore tempting to speculate that As（V） triggers mitochondrial dysfunction,which may lead to defective oxidative phosphorylation subsequently causing mitochondrial oxidative damage,which in turn induces an apoptotic mode of cell death.

关键词： Sodium arsenate Microglial BV-2 cells Mitochondrial dysfunction Oxidative phosphorylation complexes Superoxide anions Apoptosis

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Oligomerized Amyloid-<i>β</i><sub>1-40</sub>Peptide Favors Cholesterol, Oxysterol, and Fatty Acid Accumulation in Human Neuronal SK-N-BE Cells

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International Journal of Clinical Medicine 2015年第11期6卷 813-824页

作者： Amira Zarrouk thomas nury Mohamed Hammami Gérard Lizard Laboratoire de Biochimie (LR12ES05)-Lab-NAFS “Nutrition Aliments Fonctionnels et Santé Vasculaire” Faculté de Médecine Université de Monastir Monastir Tunisie Equipe “Biochimie du Peroxysome Inflammation et Métabolisme Lipidique” (EA 7270) Université de Bourgogne-Franche Comté/INSERM Dijon France

Amyloid peptide, the main component of senile plaques, is a major biological characteristic of Alzheimer’s disease (AD). The aim of the present study conducted on human neuronal SK-N-BE cells was to evaluate whether oligomerized Aβ1-40-induced cell damages was associated with lipid modifications. Under treatment with Aβ1-40 (10 - 100 μM;24 - 48 h), cell viability was recorded with the MTT test and by measuring LDH activity. Mitochondrial transmembrane potential and ATP production were assessed using flow cytometry and a luciferase-based ATP bioluminescence assay, respectively. Annexin V-CF647 staining assay for cell apoptosis detection was performed using flow cytometry. Potentially intracellular cytotoxic lipids (oxysterols: 7α-hydroxycholesterol (7α-OHC), 7β-hydroxycholesterol (7β-OHC), and 7-ketocholesterol (7KC), 24(S)-hydroxycholesterol;arachidonic acid (C20:4 n-6);VLCFAs (C22:0, C24:0, C24:6 and C26:0)) were measured using gas chromatography coupled with mass spectrometry. The cellular level of docosahexaenoic acid (C22:6 n-3), often altered in AD, was also quantified. In the presence of Aβ1-40, the percentage of MTT-positive cells decreased and was associated with an increase in LDH activity. In addition, treatment with oligomerized Aβ1-40 induced a decrease of mitochondrial transmembrane potential as well as an apoptotic cell death. Sterol analysis revealed a higher cholesterol level and a significant increase of cytotoxic oxysterols per cell (7KC + 7β-OHC), and of the [(7β-OHC + 7KC)/cholesterol] ratio, considered as a lipid peroxidation index, in Aβ1-40-treated cells. An enhancement of C20:4 n-6, C22:6 n-3 and saturated VLCFAs was also observed. Therefore, Aβ1-40-induced side effects are associated with intracellular accumulation of lipids, especially cholesterol, oxysterols (7β-OHC, 7KC), C20:4 n-6, and saturated VLCFAs, which could in turn contribute to neurotoxicity.

关键词： SK-N-BE Cells Oligomerized Aβ1-40 Cholesterol Oxysterols Very Long Chain Fatty Acids

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