Anethole exerted anticonvulsant effect in pentylenetetrazolemodel of seizure in male mice: Possible antioxidant effects

Document Type : Original Article


1 Department of Basic Sciences, Faculty of Veterinary Medicine, Shahrekord University, Shahrekord, Iran

2 Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran


Background and aims: Epilepsy is a neurological disorder causing brain dysfunctions. Treatment and control of epilepsy have long been a challenge in medical sciences. Despite the variety of current anticonvulsant drugs, research continues to discover new drugs with the highest efficacy and the lowest side effects. In the present study, the anticonvulsant effects of anethole in the seizure induction with pentylenetetrazole (PTZ) were evaluated in a mice model with respect to its possible antioxidant effects. PTZ is known to cause generalized epilepsy in animal models.
Methods: Accordingly, in this experimental study, 40 mice were divided into 5 groups; the first group received normal saline, the second group received 10 mg/kg diazepam, and the third to fifth groups were given anethole at 31.25, 62.5 and 125 mg/kg, respectively. Injections were conducted intraperitoneally for one week; then seizures were induced by the intravenous injection of 90 mg/kg PTZ. After determination of the duration of seizures in different groups, the mice were finally placed under deep anesthesia and their prefrontal cortex tissue was isolated to measure nitric oxide (NO), antioxidant capacity and malondialdehyde (MDA) concentrations.
Results: The results showed that anethole increased the delay in the onset of seizures, decreased the amount of nitrite in the brain, enhanced antioxidant capacity, and reduced MDA content in a dose-dependent manner.
Conclusion: Overall, our results indicated the anticonvulsant effects of anethole that could be mediated by inhibiting oxidative stress.


1.Amini-Khoei, H., et al., Anticonvulsant effect of minocycline on pentylenetetrazole-induced seizure in mice: involvement of nitric oxide and N-methyl-D-aspartate receptor. Canadian journal of physiology and pharmacology, 2018. 96(8): p. 742-750.
2.Bazzigaluppi, P., et al., Hungry neurons: metabolic insights on seizure dynamics. International Journal of Molecular Sciences, 2017. 18(11): p. 2269.
3.Zhao, M., et al., Focal increases in perfusion and decreases in hemoglobin oxygenation precede seizure onset in spontaneous human epilepsy. Epilepsia, 2007. 48(11): p. 2059-2067.
4.Frantseva, M., et al., Oxidative stress is involved in seizure-induced neurodegeneration in the kindling model of epilepsy. Neuroscience, 2000. 97(3): p. 431-435.
5.Shin, E.-J., et al., Role of oxidative stress in epileptic seizures. Neurochemistry international, 2011. 59(2): p. 122-137.
6.White, B.C., et al., Brain ischemia and reperfusion: molecular mechanisms of neuronal injury. Journal of the neurological sciences, 2000. 179(1-2): p. 1-33.
7.Ostadhadi, S., et al., Involvement of NMDA receptors and L-arginine/nitric oxide/cyclic guanosine monophosphate pathway in the antidepressant-like effects of topiramate in mice forced swimming test. Brain research bulletin, 2016. 122: p. 62-70.
8.Riazi, K., et al., The proconvulsant effect of sildenafil in mice: role of nitric oxide–cGMP pathway. British journal of pharmacology, 2006. 147(8): p. 935-943.
9.Bahremand, A., et al., Involvement of nitric oxide-cGMP pathway in the anticonvulsant effects of lithium chloride on PTZ-induced seizure in mice. Epilepsy research, 2010. 89(2-3): p. 295-302.
10.Modaresi, M. and A. Pouriyanzadeh, Effect of Ocimum basilicum hydro alcoholic extract against pentylenetetrazole-induced seizure in mice. Armaghane danesh, 2013. 18(8): p. 615-621.
11.Cabral, P.H.B., et al., Effects of the essential oil of Croton zehntneri and its major components, anethole and estragole, on the rat corpora cavernosa. Life sciences, 2014. 112(1-2): p. 74-81.
12.Newberne, P., et al., The FEMA GRAS assessment of trans-anethole used as a flavouring substance. Food and Chemical Toxicology, 1999. 37(7): p. 789-811.
13.Coelho‐de‐Souza, A.N., et al., Effects of the essential oil of Croton zehntneri, and its constituent estragole on intestinal smooth muscle. Phytotherapy Research: An International Journal Devoted to Medical and Scientific Research on Plants and Plant Products, 1997. 11(4): p. 299-304. Siqueira, R.J.B., et al., Cardiovascular effects of the essential oil of Croton zehntneri leaves and its main constituents, anethole and estragole, in normotensive conscious rats. Life sciences, 2006. 78(20): p. 2365-2372.
15.Sá, N., et al., Anethole reduces oxidative stress and improves in vitro survival and activation of primordial follicles. Brazilian Journal of Medical and Biological Research, 2018. 51.
16.Ritter, A.M.V., et al., Effects of anethole in nociception experimental models. Evidence-Based Complementary and Alternative Medicine, 2014. 2014.
17.Van Erum, J., D. Van Dam, and P.P. De Deyn, PTZ-induced seizures in mice require a revised Racine scale. Epilepsy & Behavior, 2019. 95: p. 51-55.
18.Benzie, I.F. and M. Devaki, The ferric reducing/antioxidant power (FRAP) assay for non-enzymatic antioxidant capacity: Concepts, procedures, limitations and applications. Measurement of antioxidant activity & capacity: recent trends and applications Wiley, New York, 2018. 11: p. 77-106.
19.Darvishi-Khezri, H., et al., The impact of silymarin on antioxidant and oxidative status in patients with β-thalassemia major: A crossover, randomized controlled trial. Complementary therapies in medicine, 2017. 35: p. 25-32.
20.Kordjazy, N., et al., Elevated level of nitric oxide mediates the anti-depressant effect of rubidium chloride in mice. European journal of pharmacology, 2015. 762: p. 411-418.
21.Hitiris, N. and M.J. Brodie, Modern antiepileptic drugs: guidelines and beyond. Current opinion in neurology, 2006. 19(2): p. 175-180.
22.Abdel-Salam, O.M., et al., Capsaicin exerts anti-convulsant and neuroprotective effects in pentylenetetrazole-induced seizures. Neurochemical research, 2020: p. 1-17.
23.Samadi-Noshahr, Z., et al., trans-Anethole attenuated renal injury and reduced expressions of angiotensin II receptor (AT1R) and TGF-β in streptozotocin-induced diabetic rats. Biochimie, 2021. 185: p. 117-127.
24.Yan, X.-l., et al., Activation of UCP2 by anethole trithione suppresses neuroinflammation after intracerebral hemorrhage. Acta Pharmacologica Sinica, 2021: p. 1-18.
25.Zhang, C., et al., Trans-anethole attenuates diet-induced nonalcoholic steatohepatitis through suppressing TGF-β-mediated fibrosis. Clinics and Research in Hepatology and Gastroenterology, 2022. 46(4): p. 101833.
26.Raman, S., M. Asle-Rousta, and M. Rahnema, Protective effect of fennel, and its major component trans-anethole against social isolation induced behavioral deficits in rats. Physiology International, 2020: p. 1-10.
27.Wang, B., et al., Neuroprotective effect of anethole against neuropathic pain induced by chronic constriction injury of the sciatic nerve in mice. Neurochemical research, 2018. 43(12): p. 2404-2422.
28.Lin, T.-K., et al., Seizure-induced oxidative stress in status epilepticus: Is antioxidant beneficial? Antioxidants, 2020. 9(11): p. 1029.
29.MacMullin, P., et al., Increase in seizure susceptibility after repetitive concussion results from oxidative stress, parvalbumin-positive interneuron dysfunction and biphasic increases in glutamate/GABA ratio. Cerebral Cortex, 2020. 30(12): p. 6108-6120.
30.Loshali, A., et al., Antiepileptic effects of antioxidant potent extract from Urtica dioica Linn. root on pentylenetetrazole and maximal electroshock induced seizure models. Heliyon, 2021. 7(2): p. e06195.
31.Restuningwiyani, S., et al., Correlation of Malondialdehyde (MDA) and C-reactive Protein (CRP) Level to Neurodevelopmental Outcome in Children After the Episode of Convulsive Type Status Epilepticus. Journal of Tropical Life Science, 2018. 8(3): p. 260573.
32.Herrera-Calderon, O., et al., Anticonvulsant effect of ethanolic extract of Cyperus articulatus L. leaves on pentylenetetrazol induced seizure in mice. Journal of traditional and complementary medicine, 2018. 8(1): p. 95-99.
33.Sarahian, N., et al., The Effects of the Peroxisome-Proliferator Activated Receptor-alpha Agonist, Fenofibrate, on the Antioxidant Capacity of the Brain in Pentylenetetrazol Kindling Seizures in Mice. Reactive Oxygen Species, 2019. 7(19): p. 47–54-47–54.
34.Bagheri, S., et al., Effect of probiotic supplementation on seizure activity and cognitive performance in PTZ-induced chemical kindling. Epilepsy & Behavior, 2019. 95: p. 43-50.
35.Faghir-Ghanesefat, H., et al., The possible role of nitric oxide pathway in pentylenetetrazole preconditioning against seizure in mice. Journal of Molecular Neuroscience, 2019. 67(3): p. 477-483.
36.Rahimi, N., et al., Nitric oxide and glutamate are contributors of anti-seizure activity of rubidium chloride: a comparison with lithium. Neuroscience letters, 2019. 708: p. 134349.
37.Haj-Mirzaian, A., et al., Protective effect of minocycline on LPS-induced mitochondrial dysfunction and decreased seizure threshold through nitric oxide pathway. European journal of pharmacology, 2019. 858: p. 172446.
38.Amini-Khoei, H., et al., Implication of nitrergic system in the anticonvulsant effects of ferulic acid in pentylenetetrazole-induced seizures in male mice. Journal of basic and clinical physiology and pharmacology, 2021.