Betaine protects cerebellum from oxidative stress following levodopa and benserazide administration in rats

Document Type : Original Article


1 Division of Biochemistry, School of Veterinary Medicine, Lorestan University, Khorram Abad, Iran

2 Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorram Abad, Iran


Objective(s): The aim of the present study was to evaluate antioxidant and methyl donor effects of betaine in cerebellum following levodopa and benserazide administration in rats.
Materials and Methods: Sprague-Dawley male rats were treated with levodopa (LD), betaine (Bet), levodopa plus betaine (LD/Bet), levodopa plus benserazide (LD/Ben), levodopa plus betaine-benserazide (LD/Bet-Ben), and the controls with vehicle for 10 consecutive days, orally.
Results: Treatment of rats with LD and benserazide significantly increased total homocysteine in plasma of the LD/Ben group when compared to the other groups. Lipid peroxidation of cerebellum increased significantly in LD-treated rats when compared to the other groups. In contrast, glutathione peroxidase activity and glutathione content in cerebellum were significantly higher in the betaine-treated rats when compared to the LD and LD/Ben groups. Serum dopamine concentration increased significantly in LD-treated rats in comparison with the LD/Ben group. LD/Bet-treated rats also demonstrated significantly higher dopamine levels when compared to the LD/Ben group.
Conclusion: We observed valuable effects of Bet in combination with LD and benserazide, which routinely were used for Parkinson’s disease (PD) treatment, in experimentally-induced oxidative stress and hyperhomocysteinemia in rats. Therefore, it seems that Bet is a vital and promising agent regarding PD for future clinical trials in humans.


1. Jenner P. Oxidative Stress in Parkinson’s Disease. Ann Neurol 2003; 53:S26–S38.
2. Ossig C, Reichmann H. Treatment of Parkinson’s disease in the advanced stage. J Neural Transm 2013; 120:523–529.
3. Basma AN, Morris EJ, Nicklas WJ, Geller HM. L-DOPA cytotoxicity to PC12 cells in culture is via auto-oxidation. J Neurochem 1995; 64:825–832.
4. Blandini F, Fancellu R, Martignoni E, Mangiagalli A, Pacchetti C, Samuele A, et al. Plasma Homocysteine and L-DOPA Metabolism in Patients with Parkinson Disease. Clin Chem 2001; 47:1102-1104.
5. Martignoni E, Blandini F, Godi L, Desideri S, Pacchetti C, Mancini F, et al. Peripheral markers of oxidative stress in Parkinson’s disease. The role of L-DOPA. Free Radic Biol Med 1999; 27:428–437.
6. Alirezaei M, Jelodar G, Niknam P, Ghayemi Z, Nazifi S. Betaine prevents ethanol-induced oxidative stress and reduces total homocysteine in the rat cerebellum. J Physiol Biochem 2011; 67: 605-612.
7. Rogers JD, Sanchez-Saffon A, Frol AB, Diaz-Arrastia R. Elevated plasma homocysteine levels in patients treated with levodopa: association with vascular disease. Arch Neurol 2003; 60: 59-64.
8. Nissinen E, Nissinen H, Larjonmaa H, Vaananen A, Helkamaa T, Reenila I, et al. The COMT inhibitor, entacapone, reduces levodopa-induced elevations in plasma homocysteine in healthy adult rats. J Neural Transm 2005; 112: 1213-1221.
9. Dal-Pizzol F, Ritter C, Cassol-Jr OJ, Rezin GT, Petronilho F, Zugno AI, et al. Oxidative mechanisms of brain dysfunction during sepsis. Neurochir Res 2010; 35:1–12.
10. Lee ES, Chen H, Soliman KF, Charlton CG. Effects of homocysteine on the dopaminergic system and behavior in rodents. Neurotoxicology 2005; 26:361-371.
11. Lutz UC .Alterations in homocysteine metabolism among alcohol dependent patients–clinical, pathobiochemical and genetic aspects. Curr Drug Abuse Rev 2008; 1:47-55.
12. Chern CL, Huang RF, Chen YH, Cheng JT, Liu TZ. Folate deficiency-induced oxidative stress and apoptosis are mediated via homocysteine-dependent overproduction of hydrogen peroxide and enhanced activation of NF-kappa B in human Hep G2 cells. Biomed Pharmacother 2001; 55: 434-42.
13. Ho PI, Ortiz D, Rogers E, Shea TB. Multiple aspects of homocysteine neurotoxicity: glutamate excitotoxicity, kinase hyperactivation and DNA damage. J Neurosci Res 2002; 70: 694-702.
14. Bleich S, Degner D, Sperling W, Bönsch D, Thürauf N, Kornhuber J. Homocysteine as a neurotoxin in chronic alcoholism. Prog Neuropsychopharmacol Biol Psychiatry 2004; 28:453-464.
15. Huang RF, Huang SM, Lin BS, Wei JS, Liu TZ. Homocysteine thiolactone induces apoptotic DNA damage mediated by increased intracellular hydrogen peroxide and caspase 3 activation in HL-60 cells. Life Sci 2001; 68:2799–2811.
16. Austin RC, Sood SK, Dorward AM, Singh G, Shaughnessy SG, Pamidi S, et al.  Homocysteine dependent alterations in mitochondrial gene expression, function and structure. Homocysteine and H2O2 act synergistically to enhance mitochondrial damage. J Biol Chem 1998; 273:30808–30817.
17. Seshadri S, Beiser A, Selhub J. Plasma homocysteine as a risk factor for dementia and Alzheimer’s disease. N Engl J Med 2002; 346:476-483.
18. Morris MS. Homocysteine and alzheimer’s disease. Lancet Neurol 2003; 2:425-428.
19. Muzzi C, Bertocci E, Terzuoli L, Porcelli B, Ciari I, Pagani R, et al. Simultaneous determination of serum concentrations of levodopa, dopamine, 3-O-methyldopa and α-methyldopa by HPLC. Biomed Pharmacother 2008; 62: 253-258.
20. Muller T, Jugel C, Ehret R, Ebersbach G, Bengel G, Muhlack S, et al. Elevation of total homocysteine levels in patients with Parkinson´s disease treated with duodenal levodopa/carbidopa gel. J Neural Transm 2011; 118:1329-1333.
21. Muller T, Erdmann C, Muhlack S, Bremen D, Przuntek H, Goetze O, et al. Pharmacokinetic behaviour of levodopa and 3-O-methyldopa after repeat administration of levodopa/carbidopa with and without entacapone in patients with Parkinson’s disease. J Neural Transm 2006; 113:1441–1448.
22. Muller T, Renger K, Kuhn W. Levodopa-associated increase of homocysteine levels and sural axonal neurodegeneration. Arch Neurol 2004; 61:657–660.
23. Schapira AH. The clinical relevance of levodopa toxicity in the treatment of Parkinson's disease. Mov Disord 2008; 23: S515-S520.
24. Alirezaei M, Jelodar G, Ghayemi Z. Antioxidant defense of betaine against oxidative stress induced by ethanol in the rat testes. Int J Pept Res Ther 2012; 18:239-247.
25. Alirezaei M, Niknam P, Jelodar G. Betaine elevates ovarian antioxidant enzyme activities and demonstrates methyl donor effect in non-pregnant rats. Int J Pept Res Ther 2012; 18:281-290.
26. Alirezaei M, Saeb M, Javidnia K, Nazifi S, Saeb S. Hyperhomocysteinemia reduction in ethanol-fed rabbits by oral betaine. Comp Clin Pathol 2012; 21:421-427.
27. Golbahar J, Aminzadeh MA, Hamidi SA, Omrani GR. Association of red blood cell 5-methyltetrahydrfoate folate with bone mineral density in postmenopausal Iranian women. Osteoporos Int 2005; 16:1894-1898.
28. Karthikeyan G, Thachil A, Sharma S, Kalaivani M, Ramakrishnan L. Elevated high sensitivity CRP levels in patients with mitral stenosis and left atrial thrombus. Int J Cardiol 2007; 122:252-254.
29. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193:265-275.
30. Subbarao KV, Richardson JS, Ang LC. Autopsy samples of Alzheimer's cortex show increased peroxidation in vitro. J Neurochem 1990; 55:342-345.
31. Sedlak J, Lindsay RH. Estimation of total, proteinbound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem 1968; 25:192–205.
32. Neamati S, Alirezaei M, Kheradmand A. Ghrelin acts as an antioxidant agent in the rat kidney. Int J Pept Res Ther 2011; 17:239-245.
33. Benson R, Crowell B Jr, Hill B, Doonquah K, Charlton C. The effects of L-Dopa on the activity of methionine adenosyltransferase: relevance to L-Dopa therapy and tolerance. Neurochem Res 1993; 18:325–330.
34. Miller JW, Shukitt-Hale B, Villalobos-Molina R, Nadeau MR, Selhub J, Joseph JA. Effect of L-Dopa and the catechol-O-methyltransferase inhibitor Ro 41-0960 on sulfur amino acid metabolites in rats. Clin Neuropharmacol 1997; 20:55–66.
35. Liu XX, Wilson K, Charlton CG. Effects of L-DOPA treatment on methylation in mouse brain: implications for the side effects of L-DOPA. Life Sci 2000; 66:2277–2288.
36. De La Cruz JP, Pavı´a J, Gonza´lez-Correa JA. Effects of chronic administration of S-adenosyl-L-methionine on brain oxidative stress in rats. Naunyn Schmiedebergs Arch Pharmacol 2000; 361:47–52.
37. Kanbak G, Arslan OC, Dokumacioglu A, Kartkaya K, Inal ME. Effects of Chronic Ethanol Consumption on Brain Synaptosomes and Protective Role of
Betaine. Neurochem Res 2008; 33:539–544.
38. Zhao WQ, Williams Z, Shepherd KR. S-adenosyl-methionine-induced apoptosis in PC12 cells. J Neurosci Res 2002; 69:519-529.
39. Sachdev PS, Valenzuela M, Brodaty H, Wang XL, Looi J, Lorentz L, et al. Homocysteine as a risk factor for cognitive impairment in stroke patients. Dement Geriatr Cogn Disord 2003; 15:155–162.
40. Haubrich DR, Gerber NH. Choline dehydrogenase. Assay, properties and inhibitors. Biochem Pharmacol 1981; 30:2993.
41. Smith AM, Zeve DR, Grisel JJ, Chen WJA. Neonatal alcohol exposure increases malondialdehyde (MDA) and glutathione (GSH) levels in the developing cerebellum. Dev Brain Res 2005; 160: 231-238.
42. Kharbanda KK, Mailliard ME, Baldwin CR, Beckenhauer HC, Sorrell MF, Tuma DJ.Betaine attenuates alcoholic steatosis by restoring phosphatidylcholine generation via the phosphatidyl-ethanolamine methyltransferase pathway. J Hepatol 2007; 46:314–321.
43. Ganesan B, Buddhan S, Anandan R, Sivakumar R, Anbinezhilan R. Antioxidant defense of betaine against isoprenaline-induced myocardial infarction in rats. Mol Biol Rep 2010; 37:1319–1327.
44. Vajragupta O, Boonyarat C, Murakami Y. A novel neuroprotective agent with antioxidant and nitric oxide synthase inhibitory action. Free Radic Res 2006; 40:685–695.
45. Wang ZJ, Liang CL, Li GM. Neuroprotective effects of arachidonic acid against oxidative stress on rat hippocampal slices. Chem Biol Interact 2006; 163:207–217.