Oligomerized Amyloid-β1-40 Peptide Favors Cholesterol, Oxysterol, and Fatty Acid Accumulation in Human Neuronal SK-N-BE Cells


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.

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Zarrouk, A. , Nury, T. , Hammami, M. and Lizard, G. (2015) Oligomerized Amyloid-β1-40 Peptide Favors Cholesterol, Oxysterol, and Fatty Acid Accumulation in Human Neuronal SK-N-BE Cells. International Journal of Clinical Medicine, 6, 813-824. doi: 10.4236/ijcm.2015.611107.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Selkoe, D. (1994) Alzheimer’s Disease: A Central Role for Amyloid. Journal of Neuropathology & Experimental Neurology, 53, 438-447.
[2] Haass, C., Schlossmacher, M.G., Hung, A.Y., Vigo-Pelfrey, C., Mellon, A., Ostaszewski, B.L., Lieberburg, I., Koo, E.H., Schenk, D., Teplow, D.B. and Selkoe, D.J. (1992) Amyloid Beta-Peptide Is Produced by Cultured Cells during Normal Metabolism. Nature, 359, 322-325.
[3] Bjõrkhem, I. (2006) Crossing the Barrier: Oxysterols as Cholesterol Transporters and Metabolic Modulators in the Brain. Journal of Internal Medicine, 260, 493-508.
[4] Grimm, M.O., Zimmer, V.C., Lehmann, J., Grimm, H.S. and Hartmann, T. (2013) The Impact of Cholesterol, DHA, and Sphingolipids on Alzheimer’s Disease. BioMed Research International, 814390.
[5] Grziwa, B., Grimm, M.O., Masters, C.L., Beyreuther, K., Hartmann, T. and Lichtenthaler, S.F. (2003) The Transmembrane Domain of the Amyloid Precursor Protein in Microsomal Membranes Is on Both Sides Shorter than Predicted. The Journal of Biological Chemistry, 278, 6803-6808.
[6] Leoni, V. and Caccia, C. (2013) 24S-Hydroxycholesterol in Plasma: A Marker of Cholesterol Turnover in Neurodegenerative Diseases. Biochimie, 95, 595-612.
[7] Wood, W.G., Li, L., Müller, W.E. and Eckert, G.P. (2014) Cholesterol as a Causative Factor in Alzheimer’s Disease: A Debatable Hypothesis. Journal of Neurochemistry, 129, 559-572.
[8] Seripa, D., D’Onofrio, G., Panza, F., Cascavilla, L., Masullo, C. and Pilotto, A. (2001) The Genetics of the Human APOE Polymorphism. Rejuvenation Research, 14, 491-500.
[9] Iuliano, L. (2011) Pathways of Cholesterol Oxidation via Non-Enzymatic Mechanisms. Chemistry and Physics of Lipids, 164, 457-468.
[10] Vaya, J. and Schipper, H.M. (2007) Oxysterols, Cholesterol Homeostasis, and Alzheimer Disease. Journal of Neurochemistry, 102, 1727-1737.
[11] Gamba, P., Guglielmotto, M., Testa, G., Monteleone, D., Zerbinati, C., Gargiulo, S., Biasi, F., Iuliano, L., Giaccone, G., Mauro, A., Poli, G., Tamagno, E. and Leonarduzzi, G. (2014) Up-Regulation of β-Amyloidogenesis in Neuron-Like Human Cells by Both 24- and 27-Hydroxycholesterol: Protective Effect of N-Acetyl-Cysteine. Aging Cell, 13, 561-572.
[12] Lütjohann, D., Papassotiropoulos, A., Bjõrkhem, I., Locatelli, S., Bagli, M., Oehring, R.D., Schlegel, U., Jessen, F., Rao, M.L., von Bergmann, K. and Heun, R. (2000) Plasma 24S-Hydroxycholesterol (Cerebrosterol) Is Increased in Alzheimer and Vascular Demented Patients. Journal of Lipid Research, 41, 195-198.
[13] Solomon, A., Leoni, V., Kivipelto, M., Besga, A., Oksengard, A., Julin, P., Svensson, L., Wahlund, L., Andreasen, N., Winblad, B., Soininen, H. and Bjorkhem, I. (2009) Plasma Levels of 24S-Hydroxycholesterol Reflect Brain Volumes in Patients without Objective Cognitive Impairments but Not with Alzheimer’s Disease. Neuroscience Letters, 462, 89-93.
[14] Zarrouk, A., Vejux, A., Mackrill, J., O’Callaghan, Y., Hammami, M., O’Brien, N. and Lizard, G. (2014) Involvement of Oxysterols in Age-Related Diseases and Ageing Processes. Ageing Research Reviews, 18, 148-162.
[15] Urano, Y., Ochiai, S. and Noguchi, N. (2013) Suppression of Amyloid-β Production by 24S-Hydroxycholesterol via Inhibition of Intracellular Amyloid Precursor Protein Trafficking. The FASEB Journal, 27, 4305-4315.
[16] Prasanthi, J.R., Huls, A., Thomasson, S., Thompson, A., Schommer, E. and Ghribi, O. (2009) Differential Effects of 24-Hydroxycholesterol and 27-Hydroxycholesterol on Beta-Amyloid Precursor Protein Levels and Processing in Human Neuroblastoma SH-SY5Y Cells. Molecular Neurodegeneration, 4, 1.
[17] Kou, J., Kovacs, G.G., Hoftberger, R., Kulik, W., Brodde, A., Forss-Petter, S., Hõnigschnabl, S., Gleiss, A., Brügger, B., Wanders, R., Just, W., Budka, H., Jungwirth, S., Fischer, P. and Berger, J. (2011) Peroxisomal Alterations in Alzheimer’s Disease. Acta Neuropathologica, 122, 271-283.
[18] Zarrouk, A., Riedinger, J.M., Ahmed, S.H., Hammami, S., Chaabane, W., Debbabi, M., Ben Ammou, S., Rouaud, O., Frih, M., Lizard, G. and Hammami, M. (2015) Fatty Acid Profiles in Demented Patients: Identification of Hexacosanoic Acid (C26:0) as a Blood Lipid Biomarker of Dementia. Journal of Alzheimer’s Disease, 44, 1349-1359.
[19] Lizard, G., Rouaud, O., Demarquoy, J., Cherkaoui-Malki, M. and Iuliano, L. (2012) Potential Roles of Peroxisomes in Alzheimer’s Disease and in Dementia of the Alzheimer’s Type. Journal of Alzheimer’s Disease, 29, 241-254.
[20] Wang, X., Wang, W., Li, L., Perry, G., Lee, H.G. and Zhu, X. (2014) Oxidative Stress and Mitochondrial Dysfunction in Alzheimer’s Disease. Biochimica et Biophysica Acta, 1842, 1240-1247.
[21] Zarrouk, A., Vejux, A., Nury, T., El Hajj, H.I., Haddad, M., Cherkaoui-Malki, M., Riedinger, J.M., Hammami, M. and Lizard, G. (2012) Induction of Mitochondrial Changes Associated with Oxidative Stress on Very Long Chain Fatty Acids (C22:0, C24:0, or C26:0)-Treated Human Neuronal Cells (SK-NB-E). Oxidative Medicine and Cellular Longevity, 2012, Article ID: 623257.
[22] Pikuleva, I.A. (2006) Cholesterol-Metabolizing Cytochromes P450. Drug Metabolism and Disposition, 34, 513-520.
[23] Smith, L.L. (1996) Review of Progress in Sterol Oxidations 1987-1995. Lipids, 3, 453-487.
[24] Ferrera, P., Mercado-Gómez, O., Silva-Aguilar, M., Valverde, M. and Arias, C. (2008) Cholesterol Potentiates Beta-Amyloid-Induced Toxicity in Human Neuroblastoma Cells: Involvement of Oxidative Stress. Neurochemical Research, 33, 1509-1517.
[25] García-Escudero, V., Martín-Maestro, P., Perry, G. and Avila, J. (2013) Deconstructing Mitochondrial Dysfunction in Alzheimer Disease. Oxidative Medicine and Cellular Longevity, 2013, Article ID: 162152.
[26] Jiang, F., Mao, Y., Liu, H., Xu, P., Zhang, L., Qian, X. and Sun, X. (2015) Magnesium Lithospermate B Protects Neurons against Amyloid β (1-42)-Induced Neurotoxicity through the NF-κB Pathway. Neurochemical Research, 40, 1954-1965.
[27] Volonté, C., Amadio, S., Cavaliere, F., D’Ambrosi, N., Vacca, F. and Bernardi, G. (2003) Extracellular ATP and Neurodegeneration. Current Drug Targets—CNS and Neurological Disorders, 2, 403-412.
[28] Barbero-Camps, E., Fernández, A., Baulies, A., Martinez, L., Fernández-Checa, J.C. and Colell, A. (2014) Endoplasmic Reticulum Stress Mediates Amyloid β Neurotoxicity via Mitochondrial Cholesterol Trafficking. American Journal of Pathology, 184, 2066-2081.
[29] Galbete, J.L., Martin, T.R., Peressini, E., Modena, P., Bianchi, R. and Forloni, G. (2000) Cholesterol Decreases Secretion of the Secreted form of Amyloid Precursor Protein by Interfering with Glycosylation in the Protein Secretory Pathway. Biochemical Journal, 348, 307-313.
[30] Refolo, L.M., Malester, B., La Francois, J., Bryant-Thomas, T., Wang, R., Tint, G.S., Sambamurti, K., Duff, K. and Pappolla, M.A. (2000) Hypercholesterolemia Accelerates the Alzheimer’s Amyloid Pathology in a Transgenic Mouse Model. Neurobiology of Disease, 7, 321-331.
[31] Colell, A., Fernández, A. and Fernández-Checa, J.C. (2009) Mitochondria, Cholesterol and Amyloid Beta Peptide: A Dangerous Trio in Alzheimer Disease. Journal of Bioenergetics and Biomembranes, 41, 417-423.
[32] Cutler, R.G., Kelly, J., Storie, K., Pedersen, W.A., Tammara, A., Hatanpaa, K., Troncoso, J.C. and Mattson, M.P. (2004) Involvement of Oxidative Stress-Induced Abnormalities in Ceramide and Cholesterol Metabolism in Brain Aging and Alzheimer’s Disease. Proceedings of the National Academy of Sciences of the United States of America, 101, 2070-2075.
[33] Zarrouk, A., Nury, T., Samadi, M., O’Callaghan, Y., Hammami, M., O’Brien, N.M., Lizard, G. and Mackrill, J.J. (2015) Effects of Cholesterol Oxides on Cell Death Induction and Calcium Increase in Human Neuronal Cells (SK-N-BE) and Evaluation of the Protective Effects of Docosahexaenoic Acid (DHA; C22:6 n-3). Steroids, 99, 238-247.
[34] Prasad, K.N., Hovland, A.R., La Rosa, F.G. and Hovland, P.G. (1998) Prostaglandins as Putative Neurotoxins in Alzheimer’s Disease. Proceedings of the Society for Experimental Biology and Medicine, 219, 120-125.
[35] Bazan, N.G. (2009) Cellular and Molecular Events Mediated by Docosahexaenoic Acid-Derived Neuroprotectin D1 Signaling in Photoreceptor Cell Survival and Brain Protection. Prostaglandins, Leukotrienes and Essential Fatty Acids, 81, 205-211.
[36] Fattahi, M.J. and Mirshafiey, A. (2014) Positive and Negative Effects of Prostaglandins in Alzheimer’s Disease. Psychiatry and Clinical Neurosciences, 68, 50-60.

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