Molecular and Cellular Biochemistry

, Volume 452, Issue 1–2, pp 199–217 | Cite as

The anxiolytic effects of atorvastatin and simvastatin on dietary-induced increase in homocysteine levels in rats

  • Natasa Mijailovic
  • Dragica Selakovic
  • Jovana Joksimovic
  • Vladimir Mihailovic
  • Jelena Katanic
  • Vladimir JakovljevicEmail author
  • Tamara Nikolic
  • Sergey Bolevich
  • Vladimir Zivkovic
  • Milica Pantic
  • Gvozden Rosic


The aim of this study was to evaluate the effects of atorvastatin and simvastatin on behavioral manifestations that followed hyperhomocysteinemia induced by special dietary protocols enriched in methionine and deficient in B vitamins (B6, B9, B12) by means of alterations in anxiety levels in rats. Simultaneously, we investigated the alterations of oxidative stress markers in rat hippocampus induced by applied dietary protocols. Furthermore, considering the well-known antioxidant properties of statins, we attempted to assess their impact on major markers of oxidative stress and their possible beneficial role on anxiety-like behavior effect in rats. The 4-week-old male Wistar albino rats were divided (eight per group) according to basic dietary protocols: standard chow, methionine-enriched, and methionine-enriched vitamins B (B6, B9, B12) deficient. Each dietary protocol (30 days) included groups with atorvastatin (3 mg/kg/day i.p.) and simvastatin (5 mg/kg/day i.p.). The behavioral testing was performed in the open field and elevated plus maze tests. Parameters of oxidative stress (index of lipid peroxidation, superoxide dismutase, catalase activity, glutathione) were determined in hippocampal tissue samples following decapitation after anesthesia. Methionine-load dietary protocols induced increased oxidative stress in rat hippocampus, which was accompanied by anxiogenic behavioral manifestations. The methionine-enriched diet with restricted vitamins B intake induced more pronounced anxiogenic effect, as well as increased oxidative stress compared to the methionine-load diet with normal vitamins B content. Simultaneous administration of statins showed beneficial effects by means of both decreased parameters of oxidative stress and attenuation of anxiety. The results obtained with simvastatin were more convincible compared to atorvastatin.


Homocysteine Atorvastatin Simvastatin Anxiety Oxidative stress Rats 











Methionine synthase






Superoxide dismutase




Open field


Elevated plus maze


Thiobarbituric acid reactive substance





This work was supported by the Faculty of Medical Sciences (JP 01/13), University of Kragujevac, Serbia.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Škovierová H, Vidomanová E, Mahmood S, Sopková J, Drgová A, Červeňová T, Halašova E, Lehotský J (2016) The molecular and cellular effect of homocysteine metabolism imbalance on human health. Int J Mol Sci 17(10):1733Google Scholar
  2. 2.
    Lu SC (2000) S-Adenosylmethionine. Int J Biochem Cell Biol 32:391–395Google Scholar
  3. 3.
    Petras M, Tatarkova Z, Kovalska M, Mokra D, Dobrota D, Lehotsky J, Drgova A (2014) Hyperhomocysteinemia as a risk factor for the neuronal system disorders. J Physio Pharmacol 65(1):15–23Google Scholar
  4. 4.
    Obeid R, Herrmann W (2006) Mechanisms of homocysteine neurotoxicity in neurodegenerative diseases with special reference to dementia. FEBS Lett 580:2994–3005Google Scholar
  5. 5.
    Reynolds EH, Carney MW, Toone BK (1984) Methylation and mood. Lancet 2:196–198Google Scholar
  6. 6.
    Türksoy N, Bilici R, Yalçıner A, Ozdemir Y, Ornek I, Tufan AE, Kara A (2014) Vitamin B12, folate, and homocysteine levels in patients with obsessive-compulsive disorder. Neuropsychiatr Dis Treat 10:1671–1675Google Scholar
  7. 7.
    Lehmann M, Gottfries C, Regland G B (1999) Identification of Cognitive impairment in the elderly: Homocysteine is an farly marker. Dement Geriatr Cogn Disord 10:12–20Google Scholar
  8. 8.
    Kahler SG, Fahey MC (2003) Metabolic disorders and mental retardation. Am J Med Genet Part 117:31–41Google Scholar
  9. 9.
    Chamberlin ME, Ubagai T, Mudd SH, Wilson WG, Leonard JV, Chou JY (1996) Demyelination of the brain is associated with methionine adenosyltransferase I/III deficiency. JCI 98(4):1021–1027Google Scholar
  10. 10.
    Oulhaj A, Refsum H, Beaumont H, Williams J, King E, Jacoby R, Smith AD (2010) Homocysteine as a predictor of cognitive decline in Alzheimer’s disease. Int J Geriatr Psychiatry 25:82–90Google Scholar
  11. 11.
    Blandini F, Fancellu R, Martignoni E, Mangiagalli A, Pacchetti C, Samuele A, Nappi G (2001) Plasma homocysteine and l-dopa metabolism in patients with Parkinson disease. Clin Chem 47(6):1102–1104Google Scholar
  12. 12.
    Hankey GJ, Eikelboom JW (2001) Homocysteine and stroke. Curr Opin Neurol 14(1):95–102Google Scholar
  13. 13.
    Obeid R, Mc Caddon A, Herrmann W (2007) The role of hyperhomocysteinemia and B vitamin deficiency in neurological and psychiatric diseases. Clin Chem Lab Med 45(12):1590–1606Google Scholar
  14. 14.
    Moustafa AA, Hewedi DH, Eissa AM, Frydecka D, Misiak B (2014) Homocysteine levels in schizophrenia and affective disorders—focus on cognition. Front Behav Neurosci 8:343Google Scholar
  15. 15.
    Gu P, DeFina LF, Leonard D, John S, Weiner MF, Brown ES (2012) Relationship between serum homocysteine levels and depressive symptoms: the Cooper Center Longitudinal Study. J Clin Psychiatr 73:691–695Google Scholar
  16. 16.
    Folstein M, Liu T, Peter I, Buell J, Arsenault L, Scott T et al (2007) The homocysteine hypothesis of depression. Am J Psychiatr 164:861–867Google Scholar
  17. 17.
    Tiemeier H, van Tuijl HR, Hofman A, Meijer J, Kiliaan AJ, Breteler MB (2002) Vitamin B12, folate, and homocysteine in depression: the Rotterdam study. Am J Psychiatr 159:2099–2101Google Scholar
  18. 18.
    Chung KH, Chiou HY, Chen YH (2017) Associations between serum homocysteine levels and anxiety and depression among children and adolescents in Taiwan. Sci Rep 7(1):8330Google Scholar
  19. 19.
    Atmaca M, Tezcan E, Kuloglu M, Kirtas O, Ustandag B (2005) Serum folate and homocysteine levels in patients with obsessive–compulsive disorder. Psychiatry Clin Neurosci 59(5):616–620Google Scholar
  20. 20.
    Levine J, Timinsky I, Vishne T et al (2008) Elevated serum homocysteine levels in male patients with PTSD. Depress Anxiety 25(11):154–157Google Scholar
  21. 21.
    Chen Z, Karaplis AC, Ackerman SL, Pogribny IP, Melnyk S, Lussier-Cacan S, Chen MF, Pai A, John SW, Smith RS et al (2001) Mice deficient in methylenetetrahydrofolate reductase exhibit hyperhomocysteinemia and decreased methylation capacity, with neuropathology and aortic lipid deposition. Hum Mol Genet 10:433–443Google Scholar
  22. 22.
    Jakubowski H, Perla-Kaján J, Finnell RH, Cabrera RM, Wang H, Gupta S, Kruger WD, Kraus JP, Shih DM (2009) Genetic or nutritional disorders in homocysteine or folate metabolism increase protein N-homocysteinylation in mice. FASEB J 23:1721–1727Google Scholar
  23. 23.
    Parsons RB, Waring RH, Ramsden DB, Williams AC (1998) In vitro effect of the cysteine metabolites homocysteic acid, homocysteine and cysteic acid upon human neuronal cell lines. Neurotoxicology 19:599–603Google Scholar
  24. 24.
    Kruman II, Culmsee C, Chan SL, Kruman Y, Guo Z, Penix L, Mattson MP (2000) Homocysteine elicits a DNA damage response in neurons that promotes apoptosis and hypersensitivity to excitotoxicity. J Neurosci 20:6920–6926Google Scholar
  25. 25.
    Zou CG, Banerjee R (2005) Homocysteine and redox signaling. Antioxid Redox Signal 7:547–559Google Scholar
  26. 26.
    Perna AF, Ingrosso D, De Santo NG (2003) Homocysteine and oxidative stress. Amino Acids 25:409–417Google Scholar
  27. 27.
    Halliwell B (2006) Oxidative stress and neurodegeneration: where are we now? J Neurochem 97(6):1634–1658Google Scholar
  28. 28.
    Weber GF (1994) The pathophysiology of reactive oxygen intermediates in the central nervous system. Med Hypotheses 43(4):223–230Google Scholar
  29. 29.
    Jara-Prado A, Ortega-Vazquez A, Martinez-Ruano L, Rios C, Santamaria A (2003) Homocysteine-induced brain lipid peroxidation: effects of NMDA receptor blockade, antioxidant treatment, and nitric oxide synthase inhibition. Neurotox Res 5(4):237–243Google Scholar
  30. 30.
    Lebel C (1991) Oxygen radicals: Common mediators of neurotoxicity. Neurotox Teratol 13:341–346Google Scholar
  31. 31.
    Herken H, Akyol O, Yilmaz HR, Tutkun H, Savas HA, Ozen ME, Kalenderoglu A, Gulec M (2006) Nitric oxide, adenosine deaminase, xanthine oxidase and superoxide dismutase in patients with panic disorder: alterations by antidepressant treatment. Hum Psychopharmacol 21:53–59Google Scholar
  32. 32.
    Herken H, Gurel A, Selek S, Armutcu F, Ozen ME, Bulut M, Kap O, Yumru M, Savas HA, Akyol O (2007) Adenosine deaminase, nitric oxide, superoxide dismutase, and xanthine oxidase in patients with major depression: impact of antidepressant treatment. Arch Med Res 38:247–252Google Scholar
  33. 33.
    Ersan S, Bakir S, Erdal Ersan E, Dogan O (2006) Examination of free radical metabolism and antioxidant defence system elements in patients with obsessive-compulsive disorder. Prog Neuropsychopharmacol Biol Psychiatr 30:1039–1042Google Scholar
  34. 34.
    Kodydkova J, Vavrova L, Zeman M, Jirak R, Macasek J, Stankova B, Tvrzicka E, Zak A (2009) Antioxidative enzymes and increased oxidative stress in depressive women. Clin Biochem 42:1368–1374Google Scholar
  35. 35.
    Ersoy MA, Selek S, Celik H, Erel O, Kaya MC, Savas HA, Herken H (2008) Role of oxidative and antioxidative parameters in etiopathogenesis and prognosis of panic disorder. Int J Neurosci 118:1025–1037Google Scholar
  36. 36.
    Selek S, Herken H, Bulut M, Ceylan MF, Celik H, Savas HA, Erel O (2008) Oxidative imbalance in obsessive compulsive disorder patients: a total evaluation of oxidant-antioxidant status. Prog Neuropsychopharmacol Biol Psychiatr 32:487–491Google Scholar
  37. 37.
    Atmaca M, Kuloglu M, Tezcan E, Ustundag B (2008) Antioxidant enzyme and malondialdehyde levels in patients with social phobia. Psychiatr Res 159:95–100Google Scholar
  38. 38.
    Galecki P, Szemraj J, Bienkiewicz M, Florkowski A, Galecka E (2009) Lipid peroxidation and antioxidant protection in patients during acute depressive episodes and in remission after fluoxetine treatment. Pharmacol Rep 61:436–447Google Scholar
  39. 39.
    Viggiano A, Viggiano E, Monda M, Ingrosso D, Perna AF, De Luca B (2012) Methionine-enriched diet decreases hippocampal antioxidant defences and impairs spontaneous behaviour and long term potentiation in rats. Brain Res 1471:66–74Google Scholar
  40. 40.
    Hrnčić D, Mikić J, Rašić-Marković A, Velimirović M, Stojković T, Obrenović R, Rankov-Petrović B, Šušić V, Djurić D, Petronijević N, Stanojlović O (2016) Anxiety-related behavior in hyperhomocysteinemia induced by methionine nutritional overload in rats: role of the brain oxidative stress. Can J Physiol Pharmacol 94(10):1074–1082Google Scholar
  41. 41.
    Hovatta I, Juhila J, Donner J (2010) Oxidative stress in anxiety and comorbid disorders. Neurosci Res 68(4):261–275Google Scholar
  42. 42.
    Rosic G, Joksimovic J, Selakovic D, Jakovljevic V, Živkovic V, Srejovic I, Djuric M, Djuric D (2018) The beneficial effects of sulfur-containing amino acids on cisplatin-induced cardiotoxicity and neurotoxicity in rodents. Curr Med Chem 25(3):391–403Google Scholar
  43. 43.
    Lakhan V (2010) Nutritional and herbal supplements for anxiety and anxiety-related disorders: systematic review. Nutr J 9:42Google Scholar
  44. 44.
    Vignes M, Maurice T, Lanté F, Nedjar M, Thethi K, Guiramand J, Récasens M (2006) Anxiolytic properties of green tea polyphenol (−)-epigallocatechin gallate (EGCG). Brain Res 1110:102–115Google Scholar
  45. 45.
    McFarlane SI, Muniyappa R, Francisco R, Sowers JR (2002) Pleiotropic effects of statins: lipid reduction and beyond. J Clin Endocrinol Metab 87(4):1451–1458Google Scholar
  46. 46.
    Murrow JR, Sher S, Ali S, Uphoff I, Patel R, Porkert M et al (2012) The differential effect of statins on oxidative stress and endothelial function: atorvastatin versus pravastatin. J Clin Lipidol 6(1):42–49Google Scholar
  47. 47.
    Van der Most PJ, Dolga AM, Nijholt IM, Luiten PGM, Eisel ULM (2009) Statins: mechanisms of neuroprotection. Prog Neurobiol 88(1):64–75Google Scholar
  48. 48.
    Mohammadi MT, Amini R, Jahanbakhsh Z, Shekarforoush S (2013) Effects of atorvastatin on the hypertension-induced oxidative stress in the rat brain. IBJ 17(3):152–157Google Scholar
  49. 49.
    ElBatsh MM (2015) Antidepressant-like effect of simvastatin in diabetic rats. Can J Physiol Pharmacol 93(8):649–656Google Scholar
  50. 50.
    Lin PY, Chang AY, Lin TK (2014) Simvastatin treatment exerts antidepressant-like effect in rats exposed to chronic mild stress. Pharmacol Biochem Behav 124:174–179Google Scholar
  51. 51.
    Can ÖD, Ulupınar E, Özkay ÜD, Yegin B, Öztürk Y (2012) The effect of simvastatin treatment on behavioral parameters, cognitive performance, and hippocampal morphology in rats fed a standard or a high-fat diet. Behav Pharmacol 23:582–592Google Scholar
  52. 52.
    Citraro R, Chimirri S, Aiello R, Gallelli L, Trimboli F, Britti D, De Sarro G, Russo E (2014) Protective effects of some statins on epileptogenesis and depressive-like behavior in WAG/Rij rats, a genetic animal model of absence epilepsy. Epilepsia 55(8):1284–1291Google Scholar
  53. 53.
    Anupama GM, Shrishail HV, Shashikant T (2013) Evaluation of antidepressant activity of simvastatin, lovastatin and atorvastatin in male swiss mice - an experimental study. Int J Drug Dev Res 5(2):102–108Google Scholar
  54. 54.
    Bjelland I, Tell G, Vollset S, Refsuem H, Ueland P (2003) Folate, vitamin B12, homocysteine, and the MTHFR 677CўT polymorphism in anxiety and depression, the Hordaland Homocysteine Study. Arch Gen Psychiatr 60(6):618–626Google Scholar
  55. 55.
    Sodha NR, Boodhwani M, Ramlawi B, Clements RT, Mieno S, Feng J, Xu SH, Bianchi S, Sellke FW (2008) Atorvastatin increases myocardial indices of oxidative stress in a porcine model of hypercholesterolemia and chronic ischemia. J Card Surg 23(4):312–320Google Scholar
  56. 56.
    Parle M, Singh N (2007) Reversal of memory deficits by atorvastatin and simvastatin in rats. Yakugaku Zasshi 127(7):1125–1137Google Scholar
  57. 57.
    Prut L, Belzung C (2003) The open field as a paradigm to measure the effect of drugs on anxiety-like behaviours: a review. Eur J Pharmacol 463(1–3):3–33Google Scholar
  58. 58.
    Pellow S, Chopin P, File SE, Briley M (1985) Validation of open:closed arm entries in an elevated plus-maze as a measure of anxiety in the rat. J Neurosci Methods 14(3):149–167Google Scholar
  59. 59.
    Pellow S, File SE (1986) Anxiolytic and anxiogenic drug effects on exploratory activity in an elevated plus-maze: a novel test of anxiety in the rat. Pharmacol Biochem Behav 24(3):525–529Google Scholar
  60. 60.
    Selakovic D, Joksimovic J, Obradovic D, Milovanovic D, Djuric M, Rosic G (2016) The adverse effects of exercise and supraphysiological dose of testosterone-enanthate (TE) on exploratory activity in elevated plus maze (EPM) test—indications for using total exploratory activity (TEA) as a new parameter for exploratory activity estimation in EPM. Neuroendocrinol Lett 37(5):101–106Google Scholar
  61. 61.
    Li KW (2011) Neuroproteomics. Humana Press Springer, New YorkGoogle Scholar
  62. 62.
    Wohlenberg M, Almeida D, Bokowski L, Medeiros N, Agostini F, Funchal C, Dani C (2014) Antioxidant activity of grapevine leaf extracts against oxidative stress induced by carbon tetrachloride in cerebral cortex, hippocampus and cerebellum of rats. Antioxidants 3(2):200–211Google Scholar
  63. 63.
    Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358Google Scholar
  64. 64.
    Misra HP, Fridovich I (1972) The role of superoxide anion in the auto-oxidation of epinephrine and simple assay for superoxide dismutase. J Biol Chem 247:3170–3175Google Scholar
  65. 65.
    Beers RF, Sizer IW (1952) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195(1):133–140Google Scholar
  66. 66.
    Ellman GL (1959) Tissue sulphydryl group. Arch Biochem Biophys 82:70–77Google Scholar
  67. 67.
    Lowry OH, Rosebrough NL, Farr AL, Randall RI (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  68. 68.
    Streck EL, Vieira PS, Wannmacher CM, Dutra-Filho CS, Wajner M, Wyse AT (2003) In vitro effect of homocysteine on some parameters of oxidative stress in rat hippocampus. Metab Brain Dis 18(2):147–154Google Scholar
  69. 69.
    Nikolić T (2017) The effects of hyperhomocysteinemia on myocardial function, coronary circulation and redox status of the isolated heart rat: role of hydroxymethyl glutaryl inhibitor coenzyme-A (HMG-COA) reductase. Dissertation, University of KragujevacGoogle Scholar
  70. 70.
    Kamath AF, Chauhan AK, Kisucka J, Dole VS, Loscalzo J, Handy DE, Wagner DD (2006) Elevated levels of homocysteine compromise blood-brain barrier integrity in mice. Blood 107(2):591–593Google Scholar
  71. 71.
    Ridker PM, Shih J, Cook TJ et al (2002) Plasma homocysteine concentration, statin therapy, and the risk of first acute coronary events. Circulation 105:1776–1779Google Scholar
  72. 72.
    Dierkes J, Luley C, Westphal S (2007) Effect of lipid-lowering and anti-hypertensive drugs on plasma homocysteine levels. Vasc Health Risk Manag 3(1):99–108Google Scholar
  73. 73.
    Jiang S, Chen Q, Venners SA, Zhong G, Hsu YH, Xing H, Wang X, Xu X (2013) Effect of simvastatin on plasma homocysteine levels and its modification by MTHFR C677T polymorphism in Chinese patients with primary hyperlipidemia. Cardiovasc Ther 31(4):27–33Google Scholar
  74. 74.
    Ludman A, Venugopal V, Yellon DM, Hausenloy DJ (2009) Statins and cardioprotection—more than just lipid lowering? Pharmacol Ther 122(1):30–43Google Scholar
  75. 75.
    Clarke AT, Johnson PC, Hall GC, Ford I, Mills PR (2016) High dose atorvastatin associated with increased risk of significant hepatotoxicity in comparison to simvastatin in UK GPRD cohort. PLoS ONE 11(3):e0151587Google Scholar
  76. 76.
    Selakovic D, Joksimovic J, Zaletel I, Puskas N, Matovic M, Rosic G (2017) The opposite effects of nandrolone decanoate and exercise on anxiety levels in rats may involve alterations in hippocampal parvalbumin-positive interneurons. PLoS ONE 12(12):e0189595Google Scholar
  77. 77.
    Hrnčić D, Rašić- Marković A, Mikić J, Demchuk G, Leković J, Šušić V, Macut D, Djurić D, Stanojlović O (2013) Anxiety-related behavior in adult rats after acute homocysteine thiolactone treatment. Clin Neurophysiol 124(7):14–15Google Scholar
  78. 78.
    Kilic FS, Ozatik Y, Kaygisiz B, Baydemir C, Erol K (2012) Acute antidepressant and anxiolytic effects of simvastatin and its mechanisms in rats. Neurosciences 17(1):39–43Google Scholar
  79. 79.
    Pemminati S, Nandini Colaco MB, Patchava D, Shivaprakash G, Sheetal Ullal D, Gopalakrishna HN, Rathnakar UP, Shenoy AK (2012) Role of statins in animal models of anxiety in Normo-cholesterolemic rats. J Pharm Res 5(7):3764–3766Google Scholar
  80. 80.
    Young-Xu Y, Chan KA, Liao JK, Ravid S, Blatt CM (2003) Long-term statin use and psychological well-being. J Am Coll Cardiol 42(4):690–697Google Scholar
  81. 81.
    Koladiya RU, Jaggi AS, Singh N, Sharma BK (2008) Ameliorative role of Atorvastatin and Pitavastatin in L-Methionine induced vascular dementia in rats. BMC Pharmacol 8:14Google Scholar
  82. 82.
    Liu W, Zhao Y, Zhang X, Ji J (2018) Simvastatin ameliorates cognitive impairments via inhibition of oxidative stress induced apoptosis of hippocampal cells through the ERK/AKT signaling pathway in a rat model of senile dementia. Mol Med Rep 17(1):1885–1892Google Scholar

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Authors and Affiliations

  • Natasa Mijailovic
    • 1
  • Dragica Selakovic
    • 1
  • Jovana Joksimovic
    • 1
  • Vladimir Mihailovic
    • 2
  • Jelena Katanic
    • 2
  • Vladimir Jakovljevic
    • 1
    • 3
    Email author
  • Tamara Nikolic
    • 1
  • Sergey Bolevich
    • 3
  • Vladimir Zivkovic
    • 1
  • Milica Pantic
    • 1
  • Gvozden Rosic
    • 1
  1. 1.Department of Physiology, Faculty of Medical SciencesUniversity of KragujevacKragujevacSerbia
  2. 2.Department of Chemistry, Faculty of ScienceUniversity of KragujevacKragujevacSerbia
  3. 3.Department of Human Pathology1st Moscow State Medical University IM SechenovMoscowRussia

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