Skip to main content

Advertisement

SpringerLink
Log in

Thymoquinone and Vitamin C Attenuates Pentylenetetrazole-Induced Seizures Via Activation of GABAB1 Receptor in Adult Rats Cortex and Hippocampus

  • Original Paper
  • Published:
NeuroMolecular Medicine Aims and scope Submit manuscript

Abstract

Epilepsy is a common neurological disorder that leads to neuronal excitability and provoke various forms of cellular reorganization in the brain. In this study, we investigate the anti-convulsant and neuroprotective effects of thymoquinone (TQ) and vitamin C against pentylenetetrazole (PTZ)-induced generalized seizures. Epileptic seizures were induced in adult rats using systemic intraperitoneal injections of PTZ (50 mg/kg) for 7 days. Animals pretreated with either TQ or vitamin C or in combination attenuated PTZ-induced seizures and mortality in rats as well neurodegeneration in the cells. Compared to PTZ, TQ and vitamin C significantly prolonged the onset of seizures (p > 0.05) as well decrease the high-grade seizures. Analysis of electroencephalogram (EEG) recordings revealed that TQ or vitamin C supplementation significantly reduced polyspike and epileptiform discharges. Epileptic seizures caused a decline in expression of gamma-aminobutyric acid B1 receptor (GABAB1R) (p > 0.05), unchanged expression of protein kinase A (PKA), decreased calcium/calmodulin-dependent protein kinase II (CaMKII) (p > 0.05) and inhibit the phosphorylation of cAMP response element-binding protein (CREB) (p > 0.05) in cortex and hippocampus, respectively, compared with control. Changes in expression of GABAB1R, CaMKII and CREB by PTZ were reversed by TQ and vitamin C supplementation. Moreover, PTZ significantly increased Bax, decreased Bcl-2 expression and finally the activation of caspase-3. TQ and vitamin C pretreatment reversed all these deleterious effects induced by PTZ. TQ and vitamin C showed anticonvulsant effects via activation of GABAB1R/CaMKII/CREB pathway and suggest a potential therapeutic role in epilepsy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Abdel-Fattah, A. M., Matsumoto, K., & Watanabe, H. (2000). Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. European Journal of Pharmacology, 400, 89–97.

    Article  CAS  PubMed  Google Scholar 

  • Akhondian, J., Kianifar, H., Raoofziaee, M., Moayedpour, A., Toosi, M. B., & Khajedaluee, M. (2011). The effect of thymoquinone on intractable pediatric seizures (pilot study). Epilepsy Research, 93, 39–43.

    Article  CAS  PubMed  Google Scholar 

  • Al-Majed, A. A., Al-Omar, F. A., & Nagi, M. N. (2006). Neuroprotective effects of thymoquinone against transient forebrain Ischemia in the rat hippocampus. European Journal of Pharmacology, 543, 40–47.

    Article  CAS  PubMed  Google Scholar 

  • Anderson, K. J., Miller, K. M., Fugaccia, I., & Scheff, S. W. (2005). Regional distribution of Fluoro-Jade B staining in the hippocampus following traumatic brain injury. Experimental Neurology, 193, 125–130.

    Article  CAS  PubMed  Google Scholar 

  • Andrade, D. M., & Minassian, B. A. (2007). Genetics of epilepsies. Expert Review of Neurotherapeutics, 7, 727–734.

    Article  CAS  PubMed  Google Scholar 

  • Bouilleret, V., Nehlig, A., Marescaux, C., & Namer, I. J. (2000). Magnetic resonance imaging follow-up of progressive hippocampal changes in a mouse model of mesial temporal lobe epilepsy. Epilepsia, 41, 642–650.

    Article  CAS  PubMed  Google Scholar 

  • Brown, J. T., Gill, C. H., Farmer, C. E., Lanneau, C., Randall, A. D., Pangalos, M. N., et al. (2003). Mechanism contributing to the exacerbated epileptiform activity in hippocampal slices of GABAB1 receptor subunit knockout mice. Epilepsy Research, 57, 121–136.

    Article  CAS  PubMed  Google Scholar 

  • Carey, K., & Fuchs, S. (2001). Pediatric pharmacology. What you need to know for the next pediatric call. Emergency Medical Services, 30, 27–34.

    CAS  PubMed  Google Scholar 

  • Cendes, F. (2005). Progressive hippocampal and extrahippocampal atrophy in drug resistant epilepsy. Current Opinion in Neurology, 18, 173–177.

    Article  PubMed  Google Scholar 

  • Chang, Y. C., Huang, A. M., Kuo, Y. M., Wang, S. T., Chang, Y. Y., & Huang, C. C. (2003). Febrile seizures impair memory and cAMP response-element binding protein activation. Annals of Neurology, 54, 706–718.

    Article  CAS  PubMed  Google Scholar 

  • Chebib, M., & Johnston, G. A. (1999). The ABC of GABA receptors: A brief review. Clinical and Experimental Pharmacology, 26, 937–940.

    Article  CAS  Google Scholar 

  • Cheng, C. Y., Su, S. Y., Tang, N. Y., Ho, T. Y., Lo, W. Y., & Hsieh, C. L. (2010). Ferulic acid inhibits nitric oxide-induced apoptosis by enhancing GABAB1 receptor expression in transient focal cerebral ischemia in rats. Acta Pharmacologica Sinica, 31, 898–899.

    Google Scholar 

  • Costello, D. J., & Delanty, N. (2004). Oxidative injury in epilepsy: Potential for antioxidant therapy? Expert Review of Neurotherapeutics, 4, 541–553.

    Article  CAS  PubMed  Google Scholar 

  • Dong, Y. U., & Rosenberg, H. C. (2004). Prolonged changes in Ca2 +/calmodulin-dependent protein kinase II after a brief pentylenetetrazol seizure; potential role in kindling. Epilepsy Research, 58, 107–117.

    Article  CAS  PubMed  Google Scholar 

  • Duncan, J. S., Sander, J. W., Sisodiya, S. M., & Walker, M. C. (2006). Adult epilepsy. Lancet, 367, 1087–1100.

    Article  PubMed  Google Scholar 

  • Engel, T., & Henshall, D. C. (2009). Apoptosis, Bcl-2 family proteins and caspases: The ABCs of seizure-damage and epileptogenesis? International Journal of Physiology, Pathophysiology and Pharmacology, 1, 97–115.

    PubMed Central  CAS  PubMed  Google Scholar 

  • Erakovic, V., Zupan, G., Varljen, J., Laginja, J., & Simonic, A. (2001). Altered activities of rat brain metabolic enzymes caused by pentylenetetrazol kindling and pentylenetetrazol-induced seizures. Epilepsy Research, 43, 165–173.

    Article  CAS  PubMed  Google Scholar 

  • Fujikawa, D. G. (2005). Prolonged seizures and cellular injury: Understanding the connection. Epilepsy & Behavior, 7(Suppl 3), S3–S11.

    Article  Google Scholar 

  • Gonzalez-Ramirez, M., Razo-Juarez, L., Sauer-Ramirez, J. L., Gonzalez-Trujano, M. E., et al. (2010). Anticonvulsive effect of vitamin C on pentylenetetrazol-induced seizures in immature rats. Pharmacology Biochemistry and Behavior, 97, 267–272.

    Article  CAS  Google Scholar 

  • Greggio, S., Rosa, R. M., Dolganov, A., Oliveira, L. M., & Menegat, F. D. (2009). NAP prevents hippocampal oxidative damage in neonatal rats subjected to hypoxia-induced seizures. Neurobiology of Disease, 36, 435–444.

    Article  CAS  PubMed  Google Scholar 

  • Haut, S. R., Bigal, M. E., & Lipton, R. B. (2006). Chronic disorders with episodic manifestations: Focus on epilepsy and migraine. Lancet Neurology, 5, 148–157.

    Article  PubMed Central  Google Scholar 

  • Haut, S. R., Velískova, J., & Moshé, S. L. (2004). Susceptibility of immature and adult brains to seizure effects. Lancet Neurology, 3, 608–617.

    Article  Google Scholar 

  • Hosford, D. A., Clark, S., Cao, Z., Wilson, W. A., Lin, F. H., Morrisett, R. A., & Huin, A. (1992). The role of GABA receptor activation in absence seizures of lethargic (lh/lh) mice. Science, 257, 398–401.

    Article  CAS  PubMed  Google Scholar 

  • Hosseinzadeh, H., & Parvardeh, S. (2004). Anticonvulsant effects of thymoquinone, the major constituent of Nigella Sativa seeds, in mice. Phytomedicine, 11, 56–64.

    Article  CAS  PubMed  Google Scholar 

  • Houghton, P. J., Zarka, R., de las Heras, B., & Hoult, J. R. (1995). Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Medica, 611, 33–36.

    Article  Google Scholar 

  • Huang, R. Q., Bell-Horner, C. L., Dibas, M. I., Covey, D. F., Drewe, J. A., & Dillon, G. H. (2001). Pentylenetetrazole-induced inhibition of recombinant gamma-aminobutyric acid type A (GABA (A)) receptors: Mechanism and site of action. Journal of Pharmacology and Experimental Therapeutics, 298, 986–995.

    CAS  PubMed  Google Scholar 

  • Ilhan, A., Gurel, A., Armutcu, F., Kamisli, S., & Iraz, M. (2005). Antiepileptogenic and antioxidant effects of Nigella sativa oil against pentylenetetrazol-induced kindling in mice. Neuropharmacology, 49, 456–464.

    Article  CAS  PubMed  Google Scholar 

  • Jancic, D., Armentia, M. L., Valor, L. M., Olivares, R., & Barco, A. (2009). Inhibition of cAMP response element-binding protein reduces neuronal excitability and plasticity, and triggers neurodegeneration. Cerebral Cortex, 19, 2535–2547.

    Article  PubMed  Google Scholar 

  • Kanter, M. (2008). Nigella sativa and derived thymoquinone prevents hippocampal neurodegeneration after chronic toluene exposure in rats. Neurochemical Research, 33, 579–588.

    Article  CAS  PubMed  Google Scholar 

  • Kulik, A., Vida, I., Fukazawa, Y., Guetg, N., Kasugai, Y., Marker, C. L., et al. (2006). Compartment dependent colocalization of Kir3.2-containing K+ channels and GABAB receptors in hippocampal pyramidal cells. Journal of Neuroscience, 26, 4289–4297.

    Article  CAS  PubMed  Google Scholar 

  • Leung, L. S., Canning, K. J., & Shen, B. (2005). Hippocampal after discharges after GABA (B)-receptor blockade in freely moving rat. Epilepsia, 46, 203–216.

    Article  CAS  PubMed  Google Scholar 

  • Li, J., Olinger, A. B., Dassow, M. S., & Abel, M. S. (2003). Up-regulation of GABA (B) receptor mRNA and protein in the hippocampus of cocaine- and lidocaine-kindled rats. Neuroscience, 118, 451–462.

    Article  CAS  PubMed  Google Scholar 

  • Liu, Z., Vergnes, M., Depaulis, A., & Marescaux, C. (1992). Involvement of intrathalamic GABAB neurotransmission in the control of absence seizures in the rat. Neuroscience, 48, 87–93.

    Article  CAS  PubMed  Google Scholar 

  • Macdonald, R. L., & Barker, J. L. (1978). Specific antagonism of GABA-mediated postsynaptic inhibition in cultured mammalian spinal cord neurons: A common mode of convulsant action. Neurology, 28, 325–330.

    Article  CAS  PubMed  Google Scholar 

  • Mousavi, S. H., Tayarani-Najaran, Z., Asghari, M., & Sadeghnia, H. R. (2010). Protective effect of Nigella sativa extract and thymoquinone on serum/glucose deprivation-induced PC12 cells death. Cellular and Molecular Neurobiology, 30, 591–598.

    Article  CAS  PubMed  Google Scholar 

  • Nagi, M. N., & Mansour, M. A. (2000). Protective effect of thymoquinone against doxorubicin–induced cardiotoxicity in rats: A possible mechanism of protection. Pharmacological Research, 41, 283–289.

    Article  CAS  PubMed  Google Scholar 

  • Naseer, M. I., Li, S., & Kim, M. O. (2009). Maternal epileptic seizure induced by pentylenetetrazol: Apoptotic neurodegeneration and decreased GABAB1 receptor expression in prenatal rat brain. Mol Brain, 2, 1–20.

    Article  Google Scholar 

  • Naseer, M. I., Ullah, I., Al-Qahtani, M. H., Karim, S., Ullah, N., Ansari, S. A., et al. (2013). Decreased GABABR expression and increased neuronal cell death in developing rat brain after PTZ-induced seizure. Neurological Sciences, 4, 497–503.

    Article  Google Scholar 

  • Oblak, A. L., Gibbs, T. T., & Blatt, G. J. (2010). Decreased GABAB receptors in the cingulate cortex and fusiform gyrus in autism. Journal of Neurochemistry, 114, 1414–1423.

    PubMed Central  CAS  PubMed  Google Scholar 

  • Patsoukis, N., Zervoudakis, G., Georgiou, C. D., Angelatou, F., Matsokis, N. A., & Panagopoulos, N. T. (2005). Thiol redox state and lipid and protein oxidation in the mouse striatum after pentylenetetrazol-induced epileptic seizure. Epilepsia, 46, 1205–1211.

    Article  CAS  PubMed  Google Scholar 

  • Pavlova, T. V., Yakovlev, A. A., Stepanichev, M. Y., Mendzheritskii, A. M., & Gulyaeva, N. V. (2004). Pentylenetetrazole kindling induces activation of caspase-3 in the rat brain. Neuroscience and Behavioral Physiology, 34, 45–47.

    Article  CAS  PubMed  Google Scholar 

  • Pi, X., Lee, J., Li, F., & Rosenber, H. C. (2004). Decreased expression of cAMP response element-binding protein gene following pentylenetetrazole seizure. Molecular Brain Research, 127, 60–67.

    Article  CAS  PubMed  Google Scholar 

  • Pin, J. P., Kniazeff, J., Binet, V., Liu, J., Maurel, D., Galvez, T., et al. (2004). Activation mechanism of the heterodimeric GABA(B) receptor. Biochemical Pharmacology, 68, 1565–1572.

    Article  CAS  PubMed  Google Scholar 

  • Racine, R. J. (1972). Modification of seizure activity by electrical stimulation: II. Motor seizure. Electroencephalography and Clinical Neurophysiology, 32, 281–294.

    Article  CAS  PubMed  Google Scholar 

  • Santos, L. F. S., Freitas, R. L. M., Xavier, S. M. L., Saldanha, G. B., & Freitas, R. M. (2008). Neuroprotective actions of vitamin C to decreased lipid peroxidation and increased catalase activity in adult rats after pilocarpine-induced seizures. Pharmacology, Biochemistry and Behavior, 89, 1–5.

    Article  CAS  PubMed  Google Scholar 

  • Sas, K., Robotka, H., Rozsa, E., Agoston, M., Szenasi, G., Gigler, G., et al. (2008). Kynurenine diminishes the ischemia-induced histological and electrophysiological deficits in the rat hippocampus. Neurobiology of Disease, 32, 302–308.

    Article  CAS  PubMed  Google Scholar 

  • Schuler, V., Luscher, C., et al. (2001). Epilepsy, hyperalgesia, impaired memory, and loss of pre- and postsynaptic GABAB responses in mice lacking GABAB(1). Neuron, 31, 47–58.

    Article  CAS  PubMed  Google Scholar 

  • Sudha, K., Ashalatha, V. R., & Anjali, R. (2001). Oxidative Stress and antioxidants in epilepsy. Clinica Chimica Acta, 303, 19–24.

    Article  CAS  Google Scholar 

  • Tu, H., Xu, C., Zhang, W., Liu, Q., Rondard, P., Pin, J. P., & Liu, J. (2010). GABAB receptor activation protects neurons from apoptosis via IGF-1 receptor transactivatio. Journal of Neuroscience, 30, 749–759.

    Article  CAS  PubMed  Google Scholar 

  • Ullah, N., Naseer, M. I., Ullah, I., Lee, H. Y., Koh, P. O., & Kim, M. O. (2011). Protective effect of pyruvate against ethanol-induced apoptotic neurodegeneration in the developing rat brain. Neuropharmacology, 61, 1248–1255.

    Article  CAS  PubMed  Google Scholar 

  • Vollenweider, F., Bendfeldt, K., Maetzler, W., Otten, U., & Nitsch, C. (2006). GABA(B) receptor expression and cellular localization in gerbil hippocampus after transient global ischemia. Neuroscience Letters, 395, 118–123.

    Article  CAS  PubMed  Google Scholar 

  • Wei, X. W., Yan, H., Xo, B. O., Wu, Y. P., Li, C., & Zhang, G. Y. (2012). Neuroprotection of co-activation of GABA receptors by preventing caspase-3 denitrosylation in KA-induced seizures. Brain Research Bulletin, 88, 617–623.

    Article  CAS  PubMed  Google Scholar 

  • Xu, K., & Stringer, J. L. (2008). Antioxidants and free radical scavengers do not consistently delay seizure onset in animal models of acute seizures. Epilepsy & Behavior, 13, 77–82.

    Article  Google Scholar 

  • Zeng, L. H., Xu, L., Rensing, N. R., Sinatra, P. M., Rothman, S. M., & Wong, M. (2007). Kainate seizures cause acute dendritic injury and actin depolymerization in vivo. Journal of Neuroscience, 27, 11604–11613.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by the Pioneer Research Center Program through the National Research Foundation of Korea funded by the Ministry of Science, ICT & Future Planning (2012-0009521).

Conflict of interest

The authors have no competing interest to declare.

Author information

Authors and Affiliations

  1. Division of Life Science (BK21 plus), College of Natural Sciences (RINS), Gyeongsang National University, Jinju, 660-701, Republic of Korea

    Ikram Ullah, Haroon Badshah, Hae Young Lee & Myeong Ok Kim

  2. Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia

    Muhammad Imran Naseer

Authors
  1. Ikram Ullah
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haroon Badshah
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Muhammad Imran Naseer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hae Young Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Myeong Ok Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Myeong Ok Kim.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ullah, I., Badshah, H., Naseer, M.I. et al. Thymoquinone and Vitamin C Attenuates Pentylenetetrazole-Induced Seizures Via Activation of GABAB1 Receptor in Adult Rats Cortex and Hippocampus. Neuromol Med 17, 35–46 (2015). https://doi.org/10.1007/s12017-014-8337-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12017-014-8337-3

Keywords

Search

Navigation