The effects of betulinic acid chronic administration on the motor, non-motor behaviors, and globus pallidus local field potential power in a rat model of hemiparkinsonism

Document Type : Original Article


1 Persian Gulf Physiology Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishpur University of Medical Sciences, Ahvaz, Iran

2 Department of Physiology, Medicine Faculty, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran


Objective(s): Parkinson’s disease (PD) is a neurodegenerative disorder involving the central nervous system associated with motor and non-motor impairments. Betulinic acid (BA) is a natural substance considered an antioxidative agent. This study aimed to investigate the therapeutic potential of BA on motor dysfunctions and globus pallidus (GP) local EEG power in a 6-hydroxydopamine (6-OHDA)-induced rat model of hemiparkinsonism.
Materials and Methods: Adult Wistar rats were categorized into different groups, containing; Sham, PD, and treated groups including different doses of BA (0.5, 5, and 10 mg/kg, IP), and L-dopa (20 mg/kg, PO, as positive control). The lesion was induced in the right medial forebrain bundle by injection of 6-OHDA (20 µg/kg). The treatment was begun just after the approved rotational test induced by apomorphine, 14 days after 6-OHDA administration. Motor behaviors such as catalepsy and stride-length and non-motor responses, including GP local EEG, were then assessed. Also, the levels of GSH, catalase, and concentration of dopamine in the brain tissue were measured. 
Results: Treatment of hemiparkinsonian rats with BA significantly improved catalepsy and stride-length (P<0.001 and P<0.01, respectively) and GP frequency bands’ powers (P<0.001). Moreover, the activities of GSH (P<0.001), catalase (P<0.001), and the concentration of dopamine (P<0.001) in the brain were increased.
Conclusion: Current results proved the potent ability of BA to scavenge free radicals and to remove oxidative agents in the brain tissue. This natural product could be considered a possible therapeutic compound for motor and non-motor disorders in PD.


1. Abrishamdar M, Farbood Y, Sarkaki A, Rashno M, Badavi M. Evaluation of betulinic acid effects on pain, memory, anxiety, catalepsy, and oxidative stress in animal model of Parkinson’s disease. Metab Brain Dis 2022; 1-16.
2. Antunes MS, Goes AT, Boeira SP, Prigol M, Jesse CR. Protective effect of hesperidin in a model of Parkinson’s disease induced by 6-hydroxydopamine in aged mice. Nutrition 2014; 30:1415-1422.
3. Antunes MS, Ladd FVL, Ladd AABL, Moreira AL, Boeira SP, Cattelan Souza L. Hesperidin protects against behavioral alterations and loss of dopaminergic neurons in 6-OHDA-lesioned mice: the role of mitochondrial dysfunction and apoptosis. Metab Brain Dis 2021; 36:153-167.
4. Arzi A, Hemmati A, Pipelzadeh M, Latifi S, Ramesh F. The effect of Selegiline and Bromocriptine in the prophylaxis of  Perphenazine- induced psedoparkinsonism in rat: A comparative study. Jundishapur J Nat Pharm Prod 2006; 1:36-40.
5. Bi Y, Xu J, Wu X, Ye W, Yuan S, Zhang L. Synthesis and cytotoxic activity of 17-carboxylic acid modified 23-hydroxy betulinic acid ester derivatives. Bioorg Med Chem Lett 2007; 17:1475-1478.
6. Bočková M, Rektor I. Impairment of brain functions in Parkinson’s disease reflected by alterations in neural connectivity in EEG studies: A viewpoint. Clin Neurophysiol 2019; 130:239-247.
7. Brown P. Abnormal oscillatory synchronization in the motor system leads to impaired movement. Curr Opin Neurobiol 2007; 17:656-664.
8. Brown P. Bad oscillations in Parkinson’s disease. Parkinson Dis Relat Disord 2006; 27-30.
9. Brown P. Oscillatory nature of human basal ganglia activity: Relationship to the pathophysiology of Parkinson’s disease. Mov Disord 2003; 18:357-363.
10. Chaudhuri KR, Schapira AH. Non-motor symptoms of Parkinson’s disease: Dopaminergic pathophysiology and treatment. Lancet Neurol 2009; 8:464-474.
11. Cheong SL, Federico S, Spalluto G, Klotz K-N, Pastorin G. The current status of pharmacotherapy for the treatment of Parkinson’s disease: transition from single-target to multitarget therapy. Drug Discov Today 2019; 24:1769-1783.
12. Dolatshahi M, Farbood Y, Sarkaki A, Mansouri SMT, Khodadadi A. Ellagic acid improves hyperalgesia and cognitive deficiency in 6-hydroxidopamine induced rat model of Parkinson’s disease. Iran J Basic Med Sci 2015; 18:38-46.
13. do Nascimento GC, Ferrari DP, Guimaraes FS, Del Bel EA, Bortolanza M, Ferreira-Junior NC. Cannabidiol increases the nociceptive threshold in a preclinical model of Parkinson’s disease. Neuropharmacology 2020; 163:107808.
14. Esposito E, Rotilio D, Di Matteo V, Di Giulio C, Cacchio M, Algeri S. A review of specific dietary antioxidants and the effects on biochemical mechanisms related to neurodegenerative processes. Neurobiol Aging 2002; 23:719-735.
15. Fleming SM, Delville Y, Schallert T. An intermittent, controlled-rate, slow progressive degeneration model of Parkinson’s disease: antiparkinson effects of Sinemet and protective effects of methylphenidate. Behav Brain Res 2005; 156:201-213.
16. Gálvez G, Recuero M, Canuet L, Del-Pozo F. Short-term effects of binaural beats on EEG power, functional connectivity, cognition, gait and anxiety in Parkinson’s disease. Int J Neural Syst 2018; 28:1750055.
17. Goes AT, Jesse CR, Antunes MS, Ladd FVL, Ladd AAL, Luchese C, et al. Protective role of chrysin on 6-hydroxydopamine-induced neurodegeneration a mouse model of Parkinson’s disease: Involvement of neuroinflammation and neurotrophins. Chem Biol Interact 2018; 279:111-120.
18. Grandi LC, Kaelin-Lang A, Orban G, Song W, Salvadè A, Stefani A, et al. Oscillatory activity in the cortex, motor thalamus and nucleus reticularis thalami in acute TTX and chronic 6-OHDA dopamine-depleted animals. Front Neurol 2018; 9:663.
19. Haddadi R, Eyvari-Brooshghalan S, Nayebi AM, Sabahi M, Ahmadi SA. Neuronal degeneration and oxidative stress in the SNc of 6-OHDA intoxicated rats; improving role of silymarin long-term treatment. Naunyn-Schmiedeb Arch Pharmacol 2020; 393:2427-2437.
20. Han C, Shen H, Yang Y, Sheng Y, Wang J, Li W, et al. Antrodia camphorata polysaccharide resists 6‐OHDA‐induced dopaminergic neuronal damage by inhibiting ROS‐NLRP3 activation. Brain Behav 2020; 10:e01824.
21. Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem 2005; 53:1841-1856.
22. Iancu R, Mohapel P, Brundin P, Paul G. Behavioral characterization of a unilateral 6-OHDA-lesion model of Parkinson’s disease in mice. Behav Brain Res 2005; 162:1-10.
23. Kaundal M, Deshmukh R, Akhtar M. Protective effect of betulinic acid against intracerebroventricular streptozotocin induced cognitive impairment and neuronal damage in rats: possible neurotransmitters and neuroinflammatory mechanism. Pharmacol Rep 2018; 70:540-548.
24. Kempf F, Brücke C, Salih F, Trottenberg T, Kupsch A, Schneider GH, et al. Gamma activity and reactivity in human thalamic local field potentials. Eur J Neurosci 2009; 29:943-953.
25. Kesh S, Kannan RR, Balakrishnan A. Naringenin alleviates 6-hydroxydopamine induced Parkinsonism in SHSY5Y cells and zebrafish model. Comp Biochem Physiol C Toxicol Pharmacol 2021; 239:108893.
26. Kim HG, Ju MS, Shim JS, Kim MC, Lee S-H, Huh Y, et al. Mulberry fruit protects dopaminergic neurons in toxin-induced Parkinson’s disease models. Br J Nutr 2010; 104:8-16.
27. Kirik D, Rosenblad C, Björklund A. Characterization of behavioral and neurodegenerative changes following partial lesions of the nigrostriatal dopamine system induced by intrastriatal 6-hydroxydopamine in the rat. Exp Neurol 1998; 15:259-277.
28. Kuruvilla KP, Nandhu M, Paul J, Paulose C. Oxidative stress-mediated neuronal damage in the corpus striatum of 6-hydroxydopamine lesioned Parkinson’s rats: Neuroprotection by Serotonin, GABA and bone marrow cells supplementation. J Neurol Sci 2013; 331:31-37.
29. Lee SY, Kim HH, Park SU. Recent studies on betulinic acid and its biological and pharmacological activity. EXCLI J 2015; 14:199-203.
30. Liang S, Zheng Y, Lei L, Deng X, Ai J, Li Y, et al. Corydalis edulis total alkaloids (CETA) ameliorates cognitive dysfunction in rat model of Alzheimer disease through regulation of the antioxidant stress and MAP2/NF-κB. J Ethnopharmacol 2020; 251:112540.
31. Little S, Brown P. The functional role of beta oscillations in Parkinson’s disease. Parkinsonism Relat Disord 2014; 20:S44-S48.
32. Lu P, Zhang C-c, Zhang X-m, Li H-g, Luo A-l, Tian Y-k, et al. Down-regulation of NOX4 by betulinic acid protects against cerebral ischemia-reperfusion in mice. Curr Med Sci 2017; 37:744-749.
33. Luo C, Huang C, Zhu L, Kong L, Yuan Z, Wen L, et al. Betulinic acid ameliorates the T-2 toxin-triggered intestinal impairment in mice by inhibiting inflammation and mucosal barrier dysfunction through the NF-κB signaling pathway. Toxins 2020; 12:794-808.
34. Mansouri MT, Farbood Y, Sameri MJ, Sarkaki A, Naghizadeh B, Rafeirad M. Neuroprotective effects of oral gallic acid against oxidative stress induced by 6-hydroxydopamine in rats. Food Chem 2013; 138:1028-1033.
35. Melgari J-M, Curcio G, Mastrolilli F, Salomone G, Trotta L, Tombini M, et al. Alpha and beta EEG power reflects L-dopa acute administration in parkinsonian patients. Front Aging Neurosci 2014; 6:302-308.
36. Navabi SP, Sarkaki A, Mansouri E, Badavi M, Ghadiri A, Farbood Y. The effects of betulinic acid on neurobehavioral activity, electrophysiology and histological changes in an animal model of the Alzheimer’s disease. Behav Brain Res 2018; 337:99-106.
37. Oh S-H, Lim S-C. A rapid and transient ROS generation by cadmium triggers apoptosis via caspase-dependent pathway in HepG2 cells and this is inhibited through N-acetylcysteine-mediated catalase upregulation. Toxicol Appl pharmacol 2006; 212:212-223.
38. Planchard MS, Samel MA, Kumar A, Rangachari V. The natural product betulinic acid rapidly promotes amyloid-β fibril formation at the expense of soluble oligomers. ACS Chem Neurosci 2012; 3:900-908.
39. Razavinasab M, Shamsizadeh A, Shabani M, Nazeri M, Allahtavakoli M, Asadi‐Shekaari M, et al. Pharmacological blockade of TRPV 1 receptors modulates the effects of 6‐OHDA on motor and cognitive functions in a rat model of Parkinson’s disease. Fundam Clin Pharmacol 2013; 27:632-640.
40. Rokosik SL, Napier TC. Pramipexole-induced increased probabilistic discounting: comparison between a rodent model of Parkinson’s disease and controls. Neuropsychopharmacology 2012; 37:1397-1408.
41. Sameri MJ, Sarkaki A, Farbood Y, Mansouri S. Motor disorders and impaired electrical power of pallidal EEG improved by gallic acid in animal model of Parkinson’s disease. Pak J Biol Sci 2011; 14:1109-1116.
42. Sarkaki A, Badavi M, Hoseiny N, Gharibnaseri M, Rahim F. Postmenopausal effects of intrastriatal estrogen on catalepsy and pallidal electroencephalogram in an animal model of Parkinson’s disease. Neuroscience 2008; 154:940-945.
43. Sarkaki A, Eidypour Z, Motamedi F, Farbood Y. Motor disturbances and thalamic electrical power of frequency bands’ improve by grape seed extract in animal model of Parkinson’s disease. Avicenna J phytomed 2012; 2:222-232.
44. Sarkaki A, Farbood Y, Dolatshahi M, Mansouri SMT, Khodadadi A. Neuroprotective effects of ellagic acid in a rat model of Parkinson’s disease. Acta Med Iran 2016; 494-502.
45. Shine J, Handojoseno A, Nguyen T, Tran Y, Naismith S, Nguyen H, et al. Abnormal patterns of theta frequency oscillations during the temporal evolution of freezing of gait in Parkinson’s disease. Clin Neurophysiol 2014; 125:569-576.
46. Souza P, Rodrigues C, Santiago A, Lucas N, Leitao GG, Galina Filho A. Anti-oxidant activity of natural compounds of Stachytarpheta cayennensis by scavenger of mitochondrial reactive oxygen species. Rev Bras Farmacogn 2011; 21:420-426.
47. Stoffers D, Bosboom J, Deijen J, Wolters EC, Berendse H, Stam C. Slowing of oscillatory brain activity is a stable characteristic of Parkinson’s disease without dementia. Brain 2007; 130:1847-1860.
48. Teixeira M, Souza C, Menezes A, Carmo M, Fonteles A, Gurgel J, et al. Catechin attenuates behavioral neurotoxicity induced by 6-OHDA in rats. Pharmacol Biochem Behav 2013; 110:1-7.
49. Tillerson JL, Caudle W, Reveron M, Miller G. Exercise induces behavioral recovery and attenuates neurochemical deficits in rodent models of Parkinson’s disease. Neuroscience 2003; 119:899-911.
50. Wu Q-H, Wang X, Yang W, Nüssler AK, Xiong L-Y, Kuča K, et al. Oxidative stress-mediated cytotoxicity and metabolism of T-2 toxin and deoxynivalenol in animals and humans: an update. Arch Toxicol 2014; 88:1309-1326.
51. Yi J, Zhu R, Wu J, Wu J, Tan Z. Ameliorative effect of betulinic acid on oxidative damage and apoptosis in the splenocytes of dexamethasone treated mice. Int Immunopharmacol 2015; 27:85-94.
52. Yi J, Zhu R, Wu J, Wu J, Xia W, Zhu L, et al. In vivo protective effect of betulinic acid on dexamethasone induced thymocyte apoptosis by reducing oxidative stress. Pharmacol Rep 2016; 68:95-100.
53. Zhao Y, Shi X, Wang J, Mang J, Xu Z. Betulinic acid ameliorates cerebral injury in middle cerebral artery occlusion rats through regulating autophagy. ACS Chem Neurosci 2021; 12:2829-2837.