Skip to main content
Log in

Aspartame and Soft Drink-Mediated Neurotoxicity in Rats: Implication of Oxidative Stress, Apoptotic Signaling Pathways, Electrolytes and Hormonal Levels

Metabolic Brain Disease Aims and scope Submit manuscript

Cite this article


A significant association between fructose corn syrup in sweetened beverages consumption and increased risk of detrimental central nervous system effects has been recently reported. We hypothesized that the aspartame and soft drink induced disturbances in energy production and endocrine function, which play a role in the induction of brain damage. Therefore, we aimed to assess the effect of aspartame and soft drink on brain function and the link between energy status in the brain, oxidative stress and molecular pathways of apoptosis. Thirty rats were randomly assigned to drink water, aspartame (240 mg/kg orally) and cola soft drinks (free access) daily for two months. Subchronic intake of aspartame and soft drink significantly disrupted the brain energy production, as indicated by inhibited serum and brain creatine kinase, specifically in soft drink-received rats. Moreover, they substantially altered serum electrolytes (increased Ca and Na, and depleted Cu, Fe, Zn and K levels), and accordingly the related hormonal status (increased T4 and PTH, and lowered T3 and aldosterone levels), particularly in soft drink-received rats reflecting brain damage. Additionally, significant increment of acetylcholine esterase activity concomitant with the reduction of antioxidant molecules (SOD, CAT, GSH-Px and GSH), and induction of malondialdehyde level are precisely indicative of oxidative brain damage. Brain mRNA transcripts of target genes showed that aspartame and soft drink induced upregulation of BAX, Casp3, P27 and Mdm2 (1.5-fold) and down-regulation of Bcl2, suggesting an activation of cellular apoptosis. Collectively, subchronic aspartame and soft drink-induced brain damage in rats may be driven via a mechanism that involves energy production disruption, electrolytes and hormonal imbalance, increased oxidative stress and activation of molecular pathway of neuronal apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1



Acetylcholine esterase


Acceptable daily intake


Analysis of variance


Bcl-2 like protein 4


B-cell lymphoma 2








Creatine kinase


Brain-type CK


Cardiac-type CK


Muscle-type CK


5,5′-dithiobis 2 nitrobenzoic acid


Reduced glutathione


Glutathione peroxidase



H2O2 :

Hydrogen peroxide




Mus musculus transformed mouse 3 T3 cell double minute 2


mitochondrial permeability transition pore


National Institutes of Health


Cyclin-dependent kinase inhibitor 1B (Cdkn1b)


Phosphate buffer saline


Pyruvate kinase


Parathyroid hormone


Standard error of means


Superoxide dismutase






  • Abhilash M, Sauganth Paul MV, Varghese MV, Nair RH (2013) Long-term consumption of aspartame and brain antioxidant defense status. Drug Chem Toxicol 36:135–140. doi:10.3109/01480545.2012.658403

    Article  CAS  PubMed  Google Scholar 

  • Adaramoye OA, Akanni OO (2016) Effects of long-term administration of aspartame on biochemical indices, lipid profile and redox status of cellular system of male rats. J Basic Clin Physiol Pharmacol 27:29–37. doi:10.1515/jbcpp-2014-0130

    Article  CAS  PubMed  Google Scholar 

  • Adrogue HJ, Madias NE (2000) Hypernatremia. N Engl J Med 342:1493–1499. doi:10.1056/NEJM200005183422006

    Article  CAS  PubMed  Google Scholar 

  • Aksenova MV et al. (1999) Oxidation of cytosolic proteins and expression of creatine kinase BB in frontal lobe in different neurodegenerative disorders. Dement Geriatr Cogn Disord 10:158–165 doi:17098

  • Al-Awwadi NA et al (2005) Extracts enriched in different polyphenolic families normalize increased cardiac NADPH oxidase expression while having differential effects on insulin resistance, hypertension, and cardiac hypertrophy in high-fructose-fed rats. J Agric Food Chem 53:151–157. doi:10.1021/jf048919f

    Article  CAS  PubMed  Google Scholar 

  • Alkhedaide A, Soliman MM, Ibrahim ZS (2016a) Carbonated soft drinks alter hepatic cytochrome P450 isoform expression in Wistar rats. Biomed Rep 5:607–612. doi:10.3892/br.2016.762

    Article  PubMed  PubMed Central  Google Scholar 

  • Alkhedaide A, Soliman MM, Salah-Eldin AE, Ismail TA, Alshehiri ZS, Attia HF (2016b) Chronic effects of soft drink consumption on the health state of Wistar rats: A biochemical, genetic and histopathological study. Mol Med Rep 13:5109–5117. doi:10.3892/mmr.2016.5199

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ashok I, Sheeladevi R (2014) Biochemical responses and mitochondrial mediated activation of apoptosis on long-term effect of aspartame in rat brain. Redox Biol 2:820–831. doi:10.1016/j.redox.2014.04.011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ashok I, Sheeladevi R, Wankhar D (2014) Long-term effect of aspartame (artificial sweetener) on ATPases, antioxidant status and histopathology in the rat brain. Free Radicals and Antioxidants. doi:10.1016/j.fra.2013.09.003

  • Baquedano E et al (2016) Increased oxidative stress and apoptosis in the hypothalamus of diabetic male mice in the insulin receptor substrate-2 knockout model. Dis Model Mech 9:573–583. doi:10.1242/dmm.023515

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Braverman ER et al (2009) Age-related increases in parathyroid hormone may be antecedent to both osteoporosis and dementia. BMC Endocr Disord 9:21. doi:10.1186/1472-6823-9-21

    Article  PubMed  PubMed Central  Google Scholar 

  • Brewer GJ, Kanzer SH, Zimmerman EA, Celmins DF, Heckman SM, Dick R (2010) Copper and ceruloplasmin abnormalities in Alzheimer's disease. Am. J. Alzheimers Dis. Other Demen 25:490–497. doi:10.1177/1533317510375083

    Article  PubMed  Google Scholar 

  • Butchko HH, Kotsonis FN (1991) Acceptable daily intake vs actual intake: the aspartame example. J Am Coll Nutr 10:258–266

    Article  CAS  PubMed  Google Scholar 

  • Cao G et al (2001) Intracellular Bax translocation after transient cerebral ischemia: implications for a role of the mitochondrial apoptotic signaling pathway in ischemic neuronal death. J Cereb Blood Flow Metab 21:321–333. doi:10.1097/00004647-200104000-00001

    Article  CAS  PubMed  Google Scholar 

  • Castro GD, Costantini MH, Delgado de Layno AM, Castro JA (2002) Rat liver microsomal and nuclear activation of methanol to hydroxymethyl free radicals. Toxicol Lett 129:227–236

    Article  CAS  PubMed  Google Scholar 

  • Chen PE et al (2005) Structural features of the glutamate binding site in recombinant NR1/NR2A N-methyl-D-aspartate receptors determined by site-directed mutagenesis and molecular modeling. Mol Pharmacol 67:1470–1484. doi:10.1124/mol.104.008185

    Article  CAS  PubMed  Google Scholar 

  • Costabeber E, Kessler A, Severo Dutra-Filho C, de Souza Wyse AT, Wajner M, Wannmacher CM (2003) Hyperphenylalaninemia reduces creatine kinase activity in the cerebral cortex of rats. Int J Dev Neurosci 21:111–116

    Article  CAS  PubMed  Google Scholar 

  • Das S, Paul S (1994) Decrease in beta-adrenergic receptors of cerebral astrocytes in hypothyroid rat brain. Life Sci 54:621–629

    Article  CAS  PubMed  Google Scholar 

  • Denehy J (2003) The health effects of soft drinks. J Sch Nurs 19:63–64. doi:10.1177/10598405030190020101

    Article  PubMed  Google Scholar 

  • Du F, Cooper A, Lukas SE, Cohen BM, Ongur D (2013) Creatine kinase and ATP synthase reaction rates in human frontal lobe measured by (3)(1)P magnetization transfer spectroscopy at 4T. Magn Reson Imaging 31:102–108. doi:10.1016/j.mri.2012.06.018

    Article  CAS  PubMed  Google Scholar 

  • Düsman E, Berti AP, Soares LC, Vicentini VEP (2013) Cytotoxicity and mutagenicity of cola and grape flavored soft drinks in bone marrow cells of rodents. Food Science and Technology (Campinas) 33:122–126

    Article  Google Scholar 

  • Dzeja PP, Terzic A (2003) Phosphotransfer networks and cellular energetics. J Exp Biol 206:2039–2047

    Article  CAS  PubMed  Google Scholar 

  • El-Terras A, Soliman MM, Alkhedaide A, Attia HF, Alharthy A, Banaja AE (2016) Carbonated soft drinks induce oxidative stress and alter the expression of certain genes in the brains of Wistar rats. Mol Med Rep 13:3147–3154. doi:10.3892/mmr.2016.4903

    Article  CAS  PubMed  Google Scholar 

  • Fernstrom JD (1989) Oral aspartame and plasma phenylalanine: pharmacokinetic difference between rodents and man, and relevance to CNS effects of phenylalanine. Short note J Neural Transm 75:159–164

    Article  CAS  PubMed  Google Scholar 

  • Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    CAS  PubMed  Google Scholar 

  • Halestrap AP, Clarke SJ, Javadov SA (2004) Mitochondrial permeability transition pore opening during myocardial reperfusion--a target for cardioprotection. Cardiovasc Res 61:372–385. doi:10.1016/S0008-6363(03)00533-9

    Article  CAS  PubMed  Google Scholar 

  • Hirasawa T, Nakamura T, Mizushima A, Morita M, Ezawa I, Miyakawa H, Kudo Y (2000) Adverse effects of an active fragment of parathyroid hormone on rat hippocampal organotypic cultures. Br J Pharmacol 129:21–28. doi:10.1038/sj.bjp.0702949

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hogervorst E, Huppert F, Matthews FE, Brayne C (2008) Thyroid function and cognitive decline in the MRC Cognitive Function and Ageing Study. Psychoneuroendocrinology 33:1013–1022. doi:10.1016/j.psyneuen.2008.05.008

    Article  CAS  PubMed  Google Scholar 

  • Horio Y, Sun Y, Liu C, Saito T, Kurasaki M (2014) Aspartame-induced apoptosis in PC12 cells. Environ Toxicol Pharmacol 37:158–165. doi:10.1016/j.etap.2013.11.021

    Article  CAS  PubMed  Google Scholar 

  • Humphries P, Pretorius E, Naude H (2008) Direct and indirect cellular effects of aspartame on the brain. Eur J Clin Nutr 62:451–462. doi:10.1038/sj.ejcn.1602866

    Article  CAS  PubMed  Google Scholar 

  • Hyder F, Patel AB, Gjedde A, Rothman DL, Behar KL, Shulman RG (2006) Neuronal-glial glucose oxidation and glutamatergic-GABAergic function. J Cereb Blood Flow Metab 26:865–877. doi:10.1038/sj.jcbfm.9600263

    Article  CAS  PubMed  Google Scholar 

  • Jaiswal S, Sharma P (2017) Role and regulation of p27 in neuronal apoptosis. J Neurochem 140:576–588. doi:10.1111/jnc.13918

    Article  CAS  PubMed  Google Scholar 

  • Kelkitli E et al (2016) Serum zinc levels in patients with iron deficiency anemia and its association with symptoms of iron deficiency anemia. Ann Hematol 95:751–756. doi:10.1007/s00277-016-2628-8

    Article  CAS  PubMed  Google Scholar 

  • Koufen P, Stark G (2000) Free radical induced inactivation of creatine kinase: sites of interaction, protection, and recovery. Biochim Biophys Acta 1501:44–50

    Article  CAS  PubMed  Google Scholar 

  • Lebda MA, El-Neweshy MS, El-Sayed YS (2012) Neurohepatic toxicity of subacute manganese chloride exposure and potential chemoprotective effects of lycopene. Neurotoxicology 33:98–104. doi:10.1016/j.neuro.2011.12.008

    Article  CAS  PubMed  Google Scholar 

  • Lebda MA, Tohamy HG, El-Sayed YS (2017) Long-term soft drink and aspartame intake induces hepatic damage via dysregulation of adipocytokines and alteration of the lipid profile and antioxidant status. Nutr Res 41:47–55. doi:10.1016/j.nutres.2017.04.002

    Article  CAS  PubMed  Google Scholar 

  • Lemasters JJ (2005) Dying a thousand deaths: redundant pathways from different organelles to apoptosis and necrosis. Gastroenterology 129:351–360

    Article  PubMed  Google Scholar 

  • Magnuson BA et al (2007) Aspartame: a safety evaluation based on current use levels, regulations, and toxicological and epidemiological studies. Crit Rev Toxicol 37:629–727. doi:10.1080/10408440701516184

    Article  CAS  PubMed  Google Scholar 

  • Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474

    Article  CAS  PubMed  Google Scholar 

  • Mohanty P, Hamouda W, Garg R, Aljada A, Ghanim H, Dandona P (2000) Glucose challenge stimulates reactive oxygen species (ROS) generation by leucocytes. J Clin Endocrinol Metab 85:2970–2973. doi:10.1210/jcem.85.8.6854

    Article  CAS  PubMed  Google Scholar 

  • Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 582:67–78

    Article  CAS  PubMed  Google Scholar 

  • Multhaup G, Schlicksupp A, Hesse L, Beher D, Ruppert T, Masters CL, Beyreuther K (1996) The amyloid precursor protein of Alzheimer's disease in the reduction of copper(II) to copper(I). Science 271:1406–1409

    Article  CAS  PubMed  Google Scholar 

  • O'Gorman E, Beutner G, Dolder M, Koretsky AP, Brdiczka D, Wallimann T (1997) The role of creatine kinase in inhibition of mitochondrial permeability transition. FEBS Lett 414:253–257

    Article  PubMed  Google Scholar 

  • O'Leary F, Hattersley L, King L, Allman-Farinelli M (2012) Sugary drink consumption behaviours among young adults at university. Nutr. Diet 69:119–123. doi:10.1111/j.1747-0080.2012.01583.x

    Article  Google Scholar 

  • Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358. doi:10.1016/0003-2697(79)90738-3

    Article  CAS  PubMed  Google Scholar 

  • Parthasarathy NJ, Kumar RS, Manikandan S, Devi RS (2006) Methanol-induced oxidative stress in rat lymphoid organs. J Occup Health 48:20–27

    Article  CAS  PubMed  Google Scholar 

  • Piñero DJ, Connor JR (2000) Iron in the Brain: An Important Contributor in Normal and Diseased States. Neuroscientist 6:435–453. doi:10.1177/107385840000600607

    Article  Google Scholar 

  • Prokic M, Paunovic M, Matic M, Djordjevic N, Ognjanovic B, Stajn A, Saicic Z (2015) Effect of aspartame on biochemical and oxidative stress parameters in rat blood. Archives of Biological Sciences 67:535–545. doi:10.2298/abs141009016p

    Article  Google Scholar 

  • Rao VK, Carlson EA, Yan SS (2014) Mitochondrial permeability transition pore is a potential drug target for neurodegeneration. Biochim Biophys Acta 1842:1267–1272. doi:10.1016/j.bbadis.2013.09.003

    Article  CAS  PubMed  Google Scholar 

  • Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973) Selenium: biochemical role as a component of glutathione peroxidase. Science 179:588–590

    Article  CAS  PubMed  Google Scholar 

  • Schlattner U, Tokarska-Schlattner M, Wallimann T (2006) Mitochondrial creatine kinase in human health and disease. Biochim Biophys Acta 1762:164–180. doi:10.1016/j.bbadis.2005.09.004

    Article  CAS  PubMed  Google Scholar 

  • Shindo A, Liang AC, Maki T, Miyamoto N, Tomimoto H, Lo EH, Arai K (2016) Subcortical ischemic vascular disease: Roles of oligodendrocyte function in experimental models of subcortical white-matter injury. J Cereb Blood Flow Metab 36:187–198. doi:10.1038/jcbfm.2015.80

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Singh M (2004) Role of micronutrients for physical growth and mental development. Indian J Pediatr 71:59–62

    Article  PubMed  Google Scholar 

  • Sinha AK (1972) Colorimetric assay of catalase. Anal Biochem 47:389–394

    Article  CAS  PubMed  Google Scholar 

  • Sullivan PG, Keller JN, Bussen WL, Scheff SW (2002) Cytochrome c release and caspase activation after traumatic brain injury. Brain Res 949:88–96

    Article  CAS  PubMed  Google Scholar 

  • Ueda S, Masutani H, Nakamura H, Tanaka T, Ueno M, Yodoi J (2002) Redox control of cell death. Antioxid Redox Signal 4:405–414. doi:10.1089/15230860260196209

    Article  CAS  PubMed  Google Scholar 

  • Wallimann T (2007) Introduction--creatine: cheap ergogenic supplement with great potential for health and disease. Subcell. Biochem 46:1–16

    Article  PubMed  Google Scholar 

  • Wang K, Wang C, Liu B, Jiang S (2010) Effect of copper deficiency on iron metabolism in rats. Wei Sheng Yan Jiu 39:426–429

    PubMed  Google Scholar 

  • Wen J, You KR, Lee SY, Song CH, Kim DG (2002) Oxidative stress-mediated apoptosis. The anticancer effect of the sesquiterpene lactone parthenolide. J Biol Chem 277:38954–38964. doi:10.1074/jbc.M203842200

    Article  CAS  PubMed  Google Scholar 

  • Wilkinson JH (1976) The principles and practice of diagnostic enzymology. E. Arnold; distributed by Year Book Medical Publishers, London

  • Yuan JS, Reed A, Chen F, Stewart CN Jr (2006) Statistical analysis of real-time PCR data. BMC Bioinf 7:85. doi:10.1186/1471-2105-7-85

    Article  Google Scholar 

  • Zlokovic BV (2011) Neurovascular pathways to neurodegeneration in Alzheimer's disease and other disorders. Nat Rev Neurosci 12:723–738. doi:10.1038/nrn3114

    CAS  PubMed  PubMed Central  Google Scholar 

Download references


All of the authors declare that they have no competing interests to disclose. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. The authors are solely responsible for the design and conduct of the study and the collection, management, analysis, and interpretation of the data, and the preparation of the manuscript. All authors had full access to the data and take responsibility for its integrity. All authors have read and agreed with the manuscript as written.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Yasser S. El-Sayed.

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lebda, M.A., Sadek, K.M. & El-Sayed, Y.S. Aspartame and Soft Drink-Mediated Neurotoxicity in Rats: Implication of Oxidative Stress, Apoptotic Signaling Pathways, Electrolytes and Hormonal Levels. Metab Brain Dis 32, 1639–1647 (2017).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: