Effect of eugenol on lithium-pilocarpine model of epilepsy: behavioral, histological, and molecular changes

Document Type : Original Article

Authors

1 School of Biology, Damghan University, Damghan, Iran

2 Institute of Biological Sciences, Damghan University, Damghan, Iran

Abstract

Objective(s): Epilepsy establishment gives rise to biochemical and morphological changes in the hippocampus. Oxidative stress, morphological changes, and mossy fiber sprouting (MFS) in the hippocampus underpin the epilepsy establishment. Eugenol is the main component of the essential oil extracted from cloves with the potential to modulate neuronal excitability. Therefore, we investigated the effect of eugenol on convulsive behavior, oxidative stress, and histological changes of the hippocampus in lithium- pilocarpine model of epilepsy.
Materials and Methods: Male Wistar rats weighing 220–250 g were divided into 4 groups; Control, Pilocarpine, Eugenol-Pilocarpine, and Eugenol. Oxidative stress markers were assayed by a biochemical method. Nissl and Timm staining were used to show neuronal survival and MFS, respectively. Behavioral convulsions were evaluated using the modified Racine scale.
Results: Eugenol decreased seizure stage and duration as well as mortality. Neuronal numbers were preserved by eugenol treatment in epileptic animals, while eugenol alone reduced the number by itself in all hippocampal sub-regions including DG, CA3, and CA1. Furthermore, eugenol alone increased MDA, GPx and SOD markers, while it increased MDA not only in combined treatment with pilocarpine but also in pilocarpine-treated animals.  In contrast to MFS enhancement in naïve animals, eugenol partially reversed the MFS enhancement induced by pilocarpine.
Conclusion: Eugenol could prevent behavioral convulsions and show neuroprotective effects through increasing neuronal survival probably by decreasing MFS and increasing the GPx antioxidant marker.

Keywords


1. Pitkanen A. Therapeutic approaches to epileptogenesis--hope on the horizon. Epilepsia 2010; 3:2-17.
2. Brodie MJ, Shorvon SD, Canger R, Halasz P, Johannessen S, Thompson P, et al. Commission on european affairs: appropriate standards of epilepsy care across Europe. ILEA. Epilepsia 1997; 38:1245-1250.
3. Kwan P, Brodie MJ. Early identification of refractory epilepsy. N Engl J Med 2000; 342:314-319.
4. Engel J, Jr. Etiology as a risk factor for medically refractory epilepsy: a case for early surgical intervention. Neurology 1998; 51:1243-1244.
5. Parent JM, Kron MM. Neurogenesis and Epilepsy. In: Noebels JL, Avoli M, Rogawski MA, Olsen RW, Delgado-Escueta AV, editors. Jasper's Basic Mechanisms of the Epilepsies. 4th ed. Bethesda (MD) 2012.
6. Paradiso B, Zucchini S, Su T, Bovolenta R, Berto E, Marconi P, et al. Localized overexpression of FGF-2 and BDNF in hippocampus reduces mossy fiber sprouting and spontaneous seizures up to 4 weeks after pilocarpine-induced status epilepticus. Epilepsia 2011; 52:572-578.
7. Buckmaster PS. Mossy Fiber Sprouting in the Dentate Gyrus. In: Noebels JL, Avoli M, Rogawski MA, Olsen RW, Delgado-Escueta AV, editors. Jasper's Basic Mechanisms of the Epilepsies. 4th ed. Bethesda (MD) 2012.
8. Castagne V, Gautschi M, Lefevre K, Posada A, Clarke PG. Relationships between neuronal death and the cellular redox status. Focus on the developing nervous system. Prog Neurobiol 1999; 59:397-423.
9. Halliwell B GJ. Free radicals in biology and medicine. 4th ed. New York: Oxford University Press; 2007.
10. Waldbaum S, Patel M. Mitochondria, oxidative stress, and temporal lobe epilepsy. Epilepsy Res 2010; 88:23-45.
11. Perry G, Nunomura A, Hirai K, Zhu X, Perez M, Avila J,      et al. Is oxidative damage the fundamental pathogenic mechanism of Alzheimer's and other neurodegenerative diseases? Free Radic Biol Med 2002; 33:1475-1479.
12. Ashrafi MR, Shams S, Nouri M, Mohseni M, Shabanian R, Yekaninejad MS, et al. A probable causative factor for an old problem: selenium and glutathione peroxidase appear to play important roles in epilepsy pathogenesis. Epilepsia 2007; 48:1750-1755.
13. Migliore L, Fontana I, Colognato R, Coppede F, Siciliano G, Murri L. Searching for the role and the most suitable biomarkers of oxidative stress in Alzheimer's disease and in other neurodegenerative diseases. Neurobiol Aging 2005; 26:587-595.
14. Bruce AJ, Baudry M. Oxygen free radicals in rat limbic structures after kainate-induced seizures. Free Radic Biol Med 1995; 18:993-1002.
15. Samuels N, Finkelstein Y, Singer SR, Oberbaum M. Herbal medicine and epilepsy: proconvulsive effects and interactions with antiepileptic drugs. Epilepsia 2008; 49:373-380.
16. Umezu T, Ito H, Nagano K, Yamakoshi M, Oouchi H, Sakaniwa M, et al. Anticonflict effects of rose oil and identification of its active constituents. Life Sci 2002; 72:91-102.
17. Cheng SS, Liu JY, Tsai KH, Chen WJ, Chang ST. Chemical composition and mosquito larvicidal activity of essential oils from leaves of different Cinnamomum osmophloeum provenances. J Agric Food Chem 2004; 52:4395-4400.
18. Bender IB. Pulpal pain diagnosis--a review. J Endod 2000; 26:175-179.
19. Koseoglu BG, Tanrikulu S, Subay RK, Sencer S. Anesthesia following overfilling of a root canal sealer into the mandibular canal: a case report. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006; 101:803-806.
20. Guenette SA, Beaudry F, Marier JF, Vachon P. Pharmacokinetics and anesthetic activity of eugenol in male Sprague-Dawley rats. J Vet Pharmacol Ther 2006; 29:265-270.
21. He M, Du M, Fan M, Bian Z. In vitro activity of eugenol against Candida albicans biofilms. Mycopathologia 2007; 163:137-143.
22. Huang CW, Chow JC, Tsai JJ, Wu SN. Characterizing the effects of Eugenol on neuronal ionic currents and hyperexcitability. Psychopharmacology (Berl) 2012; 221:575-587.
23. Gorji A, Khaleghi Ghadiri M. History of epilepsy in Medieval Iranian medicine. Neurosci Biobehav Rev 2001; 25:455-461.
24. Gorji A, Khaleghi Ghadiri M. History of headache in medieval Persian medicine. Lancet Neurol 2002; 1:510-515.
25. Aqili khorasani MS. Collection of drugs (Materia media). Tehran: Enqelab-e-Eslami Publishing and Educational Organization; 1992.
26. Zargari A. Medicinal Plants. Tehran: Tehran University press; 1990.
27. Sayyah M, Valizadeh J, Kamalinejad M. Anticonvulsant activity of the leaf essential oil of Laurus nobilis against pentylenetetrazole- and maximal electroshock-induced seizures. Phytomedicine 2002; 9:212-216.
28. Andre V, Dube C, Francois J, Leroy C, Rigoulot MA, Roch C, et al. Pathogenesis and pharmacology of epilepsy in the lithium-pilocarpine model. Epilepsia 2007; 48:41-47.
29. Garabadu D, Shah A, Ahmad A, Joshi VB, Saxena B, Palit G, et al. Eugenol as an anti-stress agent: modulation of hypothalamic-pituitary-adrenal axis and brain monoaminergic systems in a rat model of stress. Stress 2011; 14:145-155.
30. Wang D, Ren M, Guo J, Yang G, Long X, Hu R, et al. The inhibitory effects of Npas4 on seizures in pilocarpine-induced epileptic rats. PLoS One 2014; 9:e115801.
31. Li YH, Li JJ, Lu QC, Gong HQ, Liang PJ, Zhang PM. Involvement of thalamus in initiation of epileptic seizures induced by pilocarpine in mice. Neural Plasticity 2014; 2014:1-15.
32. Racine RJ. Modification of seizure activity by electrical stimulation. II. Motor seizure. Electroencephalogr Clin Neurophysiol 1972; 32:281-294.
33. Tauck DL, Nadler JV. Evidence of functional mossy fiber sprouting in hippocampal formation of kainic acid-treated rats. J Neurosci 1985; 5:1016-1022.
34. Ou HC, Chou FP, Lin TM, Yang CH, Sheu WH. Protective effects of eugenol against oxidized LDL-induced cytotoxicity and adhesion molecule expression in endothelial cells. Food Chem Toxicol 2006; 44:1485-1495.
35. Wie MB, Won MH, Lee KH, Shin JH, Lee JC, Suh HW, et al. Eugenol protects neuronal cells from excitotoxic and oxidative injury in primary cortical cultures. Neurosci Lett 1997; 225:93-96.
36. Muller M, Pape HC, Speckmann EJ, Gorji A. Effect of eugenol on spreading depression and epileptiform discharges in rat neocortical and hippocampal tissues. Neuroscience 2006; 140:743-751.
37. Jeong KH, Lee DS, Kim SR. Effects of eugenol on granule cell dispersion in a mouse model of temporal lobe epilepsy. Epilepsy Res 2015; 115:73-76.
38. Chen S, Su H, Yue C, Remy S, Royeck M, Sochivko D, et al. An increase in persistent sodium current contributes to intrinsic neuronal bursting after status epilepticus. J Neurophysiol 2011; 105:117-129.
39. Su H, Sochivko D, Becker A, Chen J, Jiang Y, Yaari Y, et al. Upregulation of a T-type Ca2+ channel causes a long-lasting modification of neuronal firing mode after status epilepticus. J Neurosci 2002; 22:3645-3655.
40. Yaari Y, Yue C, Su H. Recruitment of apical dendritic T-type Ca2+ channels by backpropagating spikes underlies de novo intrinsic bursting in hippocampal epileptogenesis. J Physiol 2007; 580:435-450.
41. Becker AJ, Pitsch J, Sochivko D, Opitz T, Staniek M, Chen CC, et al. Transcriptional upregulation of Cav3.2 mediates epileptogenesis in the pilocarpine model of epilepsy. J Neurosci 2008; 28:13341-13353.
42. Graef JD, Nordskog BK, Wiggins WF, Godwin DW. An acquired channelopathy involving thalamic T-type Ca2+ channels after status epilepticus. J Neurosci 2009; 29:4430-4441.
43. Cheung ZH, Chin WH, Chen Y, Ng YP, Ip NY. Cdk5 is involved in BDNF-stimulated dendritic growth in hippocampal neurons. PLoS Biol 2007; 5:e63.
44. Aoshima H, Hamamoto K. Potentiation of GABAA receptors expressed in Xenopus oocytes by perfume and phytoncid. Biosci Biotechnol Biochem 1999; 63:743-748.
45. Won MH, Lee JC, Kim YH, Song DK, Suh HW, Oh YS, et al. Postischemic hypothermia induced by eugenol protects hippocampal neurons from global ischemia in gerbils. Neurosci Lett 1998; 254:101-104.
46. Liu Z, Niu W, Yang X, Wang Y. Effects of combined acupuncture and eugenol on learning-memory ability and antioxidation system of hippocampus in Alzhei-mer disease rats via olfactory system stimulation. J Tradit Chin Med 2013; 33:399-402.
47. Sartorius T, Peter A, Schulz N, Drescher A, Bergheim I, Machann J, et al. Cinnamon extract improves insulin sensitivity in the brain and lowers liver fat in mouse models of obesity. PLoS One 2014; 9:e92358.
48. Walling SG, Rigoulot MA, Scharfman HE. Acute and chronic changes in glycogen phosphorylase in hippo-campus and entorhinal cortex after status epilepticus in the adult male rat. Eur J Neurosci 2007; 26:178-189.
49. Vidhya N, Devaraj SN. Induction of apoptosis by eugenol in human breast cancer cells. Indian J Exp Biol 2011; 49:871-878.
50. Pramod K, Ansari SH, Ali J. Eugenol: a natural compound with versatile pharmacological actions. Nat Prod Commun 2010; 5:1999-2006.
51. Alcantara S, Ruiz M, D'Arcangelo G, Ezan F, de Lecea L, Curran T, et al. Regional and cellular patterns of reelin mRNA expression in the forebrain of the developing and adult mouse. J Neurosci 1998; 18:7779-7799.
52. Rice DS, Curran T. Role of the reelin signaling pathway in central nervous system development. Annu Rev Neurosci 2001; 24:1005-1039.
53. Soriano E, Del Rio JA. The cells of cajal-retzius: still a mystery one century after. Neuron 2005; 46:389-394.
54. Cooper JA. A mechanism for inside-out lamination in the neocortex. Trends Neurosci 2008; 31:113-119.
55. Guilhem D, Dreyfus PA, Makiura Y, Suzuki F, Onteniente B. Short increase of BDNF messenger RNA triggers kainic acid-induced neuronal hypertrophy in adult mice. Neuroscience 1996; 72:923-931.
56. Ringstedt T, Linnarsson S, Wagner J, Lendahl U, Kokaia Z, Arenas E, et al. BDNF regulates reelin expression and Cajal-Retzius cell development in the cerebral cortex. Neuron 1998; 21:305-315.
57. Irie Y. Effects of eugenol on the central nervous system: its possible application to treatment of Alzheimer's disease, depression, and Parkinson's isease. Curr. Bioactive Compound; 2006; 2:57-66.
58. Simo S, Pujadas L, Segura MF, La Torre A, Del Rio JA, Urena JM, et al. Reelin induces the detachment of postnatal subventricular zone cells and the expression of the Egr-1 through Erk1/2 activation. Cereb Cortex 2007; 17:294-303.