Download PDF
Drug Res (Stuttg) 2013; 63(06): 311-318
DOI: 10.1055/s-0033-1337940
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Combination Therapy of Piperine and Phenytoin in Maximal Electroshock Induced Seizures in Mice: Isobolographic and Biochemical Analysis

P. Saraogi
1   Neurobehavioral Pharmacology Laboratory, Department of Pharmacology, Faculty of Pharmacy, Jamia Hamdard (Hamdard University), New Delhi, India
,
D. Vohora
1   Neurobehavioral Pharmacology Laboratory, Department of Pharmacology, Faculty of Pharmacy, Jamia Hamdard (Hamdard University), New Delhi, India
,
R. Khanam
1   Neurobehavioral Pharmacology Laboratory, Department of Pharmacology, Faculty of Pharmacy, Jamia Hamdard (Hamdard University), New Delhi, India
,
K. K. Pillai
1   Neurobehavioral Pharmacology Laboratory, Department of Pharmacology, Faculty of Pharmacy, Jamia Hamdard (Hamdard University), New Delhi, India
› Author Affiliations
Further Information

Publication History

received 29 January 2013

accepted 18 February 2013

Publication Date:
25 March 2013 (online)

Also available at
    Permissions and Reprints

Abstract

The present study was aimed to characterize the anticonvulsant effects of piperine in combination with well established antiepileptic drug (AED) phenytoin, in the mouse maximal electroshock (MES)-induced seizure model by using the type I isobolographic analysis for non-parallel dose-response relationship curves (DRRCs). Potential adverse-effect profiles of interactions of phenytoin with piperine at the fixed-ratio of 1:1 from the MES test with respect to long-term memory and skeletal muscular strength were evaluated along with free plasma concentration of piperine and phenytoin. Parameters of oxidative stress (glutathione, malondialdehyde), brain serotonin and serum calcium levels were also determined to probe the mechanism involved in the interaction. Test of parallelism revealed that 2 drugs were associated with non-parallel dose response effects, hence only one fixed ratio combination (1:1) was evaluated which displayed additive interaction between the 2 drugs with a slight tendency towards superadditivity. Free plasma concentrations of piperine and phenytoin revealed no significant changes in their concentrations when the drugs were combined at the fixed-ratio of 1:1. In combination, neither long-term memory nor skeletal muscular strength was impaired. Analysis of biochemical parameters showed that the piperine alone or in combination with phenytoin successfully reversed the parameters of oxidative stress and increased brain serotonin levels as compared to MES group. However, no significant alteration in the serum calcium levels was observed by any treatment. In conclusion, the combination displayed additive interaction and slight tendency towards synergistic potential with protection towards side effects associated with AED therapy and is worthy of consideration for further investigations.

Key words

piperine - phenytoin - maximal electroshock seizures - isobologram - oxidative stress - brain serotonin
 

  • References

  • 1 Luszczki JJ, Wu ZJ, Rasjewski G et al. Isobolographic characterisation of interactions of retigabine with carbamazepine, lamotrigine, and valproate in the mouse maximal electroshock-induced seizure model. Naunyn-Schmiedeberg’s Arch Pharmacol 2009; 379: 163-179
    PubMedGoogle Scholar
  • 2 Loscher W, Schmidt D. New Horizons in the development of antiepileptic drugs: the search for new targets. Epilepsy Res 2004; 60: 77-159
    CrossrefPubMedGoogle Scholar
  • 3 Rowan AJ, Meijer JW, de Beer-Pawlikowski N, van der Geest P, Meinardi H. Valproate-ethosuximide combination therapy for refractory absence seizures. Arch Neurol 1983; 40: 797-802
    CrossrefPubMedGoogle Scholar
  • 4 Ferrie CD, Robinson RO, Knott C et al. Lamotrigine as an add-on drug in typical absence seizures. Acta Neurol Scand 1995; 91: 200-202
    PubMedGoogle Scholar
  • 5 Leach JP, Brodie MJ. Synergism with GABAergic drugs in refractory epilepsy. Lancet 1994; 343: 314-319
    PubMedGoogle Scholar
  • 6 Stephen LJ, Sills GJ, Brodie MJ. Lamotrigine and topiramate may be useful combination. Lancet 1998; 315: 958-959
    PubMedGoogle Scholar
  • 7 D’Hooge R, Pei YQ, Raes A et al. Anticonvulsant activity of piperine on seizures induced by excitatory amino acid receptor agonists. Arzneimittelforschung 1996; 46: 557-560
    PubMedGoogle Scholar
  • 8 Li S, Wang C, Li W et al. Antidepressant-like effects of piperine and its derivative, Antiepilepsirine. J Asian Nat Prod Res 2007; 9: 435-444
    PubMedGoogle Scholar
  • 9 Li S, Wang C, Wang M et al. Antidepressant like effects of piperine in chronic mild stress treated mice and its possible mechanisms. Life Sci 2007; 80: 1373-1381
    CrossrefPubMedGoogle Scholar
  • 10 Bai YF, Xu H. Protective action of piperine against experimental gastric ulcer. Acta Pharmacol Sin 2000; 21: 357-359
    PubMedGoogle Scholar
  • 11 Selvendiran K, Singh JP, Krishnan KB et al. Cytoprotective effect of piperine against benzoapyrene induced lung cancer with reference to lipid peroxidation and antioxidant system in Swiss albino mice. Fitoterapia 2003; 74: 109-115
    CrossrefPubMedGoogle Scholar
  • 12 Nsour WM, Lau CB, Wong IC. Review on phytotherapy in epilepsy. Seizure 9: 96-107
    PubMedGoogle Scholar
  • 13 Chonpathompikunlert P, Wattanathorn J, Muchimapura S. Piperine, the main alkaloid of Thai black pepper, protects against neurodegeneration and cognitive impairment in animal model of cognitive deficit like condition of Alzheimer’s disease. Food Chem Toxicol 2010; 48: 798-802
    PubMedGoogle Scholar
  • 14 Sharma V, Nehru B, Munshi A et al Antioxidant potential of curcumin against oxidative insult induced by pentylenetetrazol in epileptic rats. Methods Find Exp Clin Pharmacol 2010; 32: 227-232
    CrossrefPubMedGoogle Scholar
  • 15 Fu M, Sun ZH, Zuo HC. Neuroprotective effect of piperine on primarily cultured hippocampal neurons. Biol Pharm Bull 2010; 33: 598-603
    CrossrefPubMedGoogle Scholar
  • 16 Zaugg J, Baburin I, Strommer B et al. HPLC-based activity profiling: discovery of piperine as a positive GABA(A) receptor modulator targeting a benzodiazepine-independent binding site. J Nat Prod 2010; 73: 185-191
    CrossrefPubMedGoogle Scholar
  • 17 Shrivastava P, Vaibhav K, Tabassum R et al. Anti-apoptotic and anti-inflammatory effect of Piperine on 6-OHDA induced Parkinson’s rat model. J Nutr Biochem DOI: dx.doi.org/10.1016/j.jnutbio.2012.03.018.
    PubMedGoogle Scholar
  • 18 Lv XC, Huang HP, Lin WH et al. Effect of serotonin depletion on seizures learning-memory in pilocarpine-induced epileptic rats. Zhongguo Ying Yong Sheng Li Xue Za Zhi 2012; 28: 210-213
    PubMedGoogle Scholar
  • 19 Lasoń W, Dudra-Jastrzębska M, Rejdak K et al. Basic mechanisms of antiepileptic drugs and their pharmacokinetic/pharmacodynamic interactions: an update. Pharmacol Rep 2011; 63: 271-292
    PubMedGoogle Scholar
  • 20 Acharya MM, Hattiangady B, Shetty AK. Progress in neuroprotective strategies for preventing epilepsy. Prog Neurobiol 2008; 84: 363-404
    CrossrefPubMedGoogle Scholar
  • 21 Wagner H, Ulrich-Merzenich G. Synergy research: Approaching a new generation of phytopharmaceuticals. Phytomedicine 2009; 16: 97-110
    CrossrefPubMedGoogle Scholar
  • 22 Patel JA, Shah US. Hepatoprotective activity of Piper longum traditional milk extract on carbon tetrachloride induced liver toxicity in Wistar rats. Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas 2009; 8: 121-129
    PubMedGoogle Scholar
  • 23 Velpandian T, Jasuja R, Bharadwaj RK et al. Piperine in food: Interference in the pharmacokinetics of phenytoin. Eur J Drug Metab Pharmacokinet 2001; 26: 241-248
    CrossrefPubMedGoogle Scholar
  • 24 Litchfield JT, Wilcoxon F. A simplified method of evaluating dose-effect experiments. J Pharmacol Exp Ther 1949; 96: 99-113
    PubMedGoogle Scholar
  • 25 Luszczki JJ. Isobolographic analysis on interaction between drugs with nonparallel dose-response relationship curves: A practical application. Naunyn-Schmiedeberg’s Arch Pharmacol 2007; 375: 105-114
    PubMedGoogle Scholar
  • 26 Devi PU, Saraogi P, Manocha A et al. Pharmacological and biochemical analysis of interactions between N-acetylcysteine and some antiepileptic drugs on experimental seizures in mice. CNS Neurosci Thera 2012; 18: 406-413
    PubMedGoogle Scholar
  • 27 Meyer OA, Tilson HA, Byrd WC et al. A method for the routine assessment of fore and hindlimb grip strength of rats and mice. Neurobehavioral Toxicol 1979; 1: 233-236
    PubMedGoogle Scholar
  • 28 Vaglenova J, Birru S, Pandiella NM et al. An assessment of the long-term developmental and behavioural teratogenicity of prenatal nicotine exposure. Behav Brain Res 2004; 150: 159-170
    CrossrefPubMedGoogle Scholar
  • 29 Bajad S, Singla AK, Bedi KL. Liquid chromatographic method for determination of piperine in rat plasma: application to pharmacokinetics. J Chromatogr B 2002; 77: 245-249
    PubMedGoogle Scholar
  • 30 Al-Humayyd MS. Diltiazem and verapamil elevate phenytoin concentrations in the rat. Int J Pharm 1996; 131: 153-158
    CrossrefPubMedGoogle Scholar
  • 31 Ellman GL. Tissue sulphydryl groups. Arch Biochem Biophys 1959; 82: 70-73
    CrossrefPubMedGoogle Scholar
  • 32 Ohkawa H, Ohishi N, Yagi K. Assay of lipid peroxides in animal tissue by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351-358
    CrossrefPubMedGoogle Scholar
  • 33 Pollard FH, Martin JV. The spectrophotometric determination of the alkaline-earth metals with murexide, eriochrome black T and with o-cresolphthalein complexone. Analyst 1956; 81: 348-353
    CrossrefPubMedGoogle Scholar
  • 34 Maickel RP, Raymond HC, Jean JR et al. PM. A Method for the determination of serotonin and norepinephrine in discrete areas of rat brain. Int J Neuropharmacol 1968; 7: 275-228
    CrossrefPubMedGoogle Scholar
  • 35 Luszczki JJ, Andres MM, Czuczwar SJ. Synergistic interaction of gabapentin and oxcarbazepine in the mouse maximal electroshock seizure model-an isobolographic analysis. Eur J Pharmacol 2005; 515: 54-61
    CrossrefPubMedGoogle Scholar
  • 36 Tallarida RJ. The interaction index: a measure of drug synergism. Pain 2002; 98: 163-168
    CrossrefPubMedGoogle Scholar
  • 37 Arya R, Gulati S, Kabra M et al. Folic acid supplementation prevents phenytoin-induced gingival overgrowth in children. Neurology 2011; 76: 1338-1343
    CrossrefPubMedGoogle Scholar
  • 38 Meier C, Kraenzlin ME. Antiepileptics and bone health. Ther Adv in Musculoskelet Dis 2011; 3: 235-243
    PubMedGoogle Scholar
  • 39 Reeta KH, Mehla J, Pahuja M et al. Pharmacokinetic and pharmacodynamic interactions of valproate, phenytoin, phenobarbitone and carbamazepine with curcumin in experimental models of epilepsy in rats. Pharmacol Biochem Behav 2011; 99: 399-407
    CrossrefPubMedGoogle Scholar
  • 40 Björnsson E. Hepatotoxicity associated with antiepileptic drugs. Acta Neurol Scand 2008; 118: 281-290
    CrossrefPubMedGoogle Scholar
  • 41 Makhov P, Golovine K, Canter D et al Co-administration of piperine and docetaxel results in improved anti-tumor efficacy via inhibition of CYP3A4 activity. Prostate 2012; 72: 661-667
    CrossrefPubMedGoogle Scholar
  • 42 Shin EJ, Jeong JH, Chung YH et al. Role of oxidative stress in epileptic seizures. Neurochem Int 2011; 59: 122-137
    CrossrefPubMedGoogle Scholar
  • 43 Wong-Ekkabut J, Xu Z, Triampo W et al. Effect of lipid peroxidation of lipid bilayers: a molecular dynamic study. Biophysics J 2007; 93: 4225-4236
    CrossrefPubMedGoogle Scholar
  • 44 Pahuja M, Mehla J, Reeta KH et al. Hydroalcoholic extract of Zizyphus jujuba ameliorates seizures, oxidative stress, and cognitive impairment in experimental models of epilepsy in rats. Epilepsy Behav 2011; 21: 356-363
    CrossrefPubMedGoogle Scholar
  • 45 Reeta KH, Mehla J, Gupta YK. Curcumin is protective against phenytoin-induced cognitive impairment and oxidative stress in rats. Brain Res 2009; 1301: 52-60
    CrossrefPubMedGoogle Scholar
  • 46 Oladipo OO, Lesi FE, Ezeaka VC. Plasma magnesium and calcium levels in children with epilepsy in lagos. Niger Postgrad Med J 2007; 14: 26-29
    PubMedGoogle Scholar
  • 47 Babayigit A, Dirik E, Bober E et al. Adverse effects of antiepileptic drugs on bone mineral density. Pediatr Neurol 2006; 35: 177-181
    CrossrefPubMedGoogle Scholar
  • 48 Richerson GB, Buchanan GF. The serotonin axis: Shared mechanism in seizures, depression, and SUDEP. Epilepsia 2011; 52: 28-38
    PubMedGoogle Scholar
  • 49 Dailey JW, Reigel CE, Mishra PK et al. Neurobiology of seizure predisposition in the genetically epilepsy-prone rat. Epilepsy Res 1989; 3: 3-17
    CrossrefPubMedGoogle Scholar
  • 50 Brennan TJ, Seeley WW, Kilgard M et al. Sound-induced seizures in serotonin 5-HT2c receptor mutant mice. Nat Genet 1997; 16: 387-390
    CrossrefPubMedGoogle Scholar
  • 51 Toczek MT, Carson RE, Lang L et al. PET imaging of 5-HT1A receptor binding in patients with temporal lobe epilepsy. Neurology 2003; 60: 749-756
    CrossrefPubMedGoogle Scholar
  • 52 Ahmad S, Fowler LJ, Whitton PS. Lamotrigine, carbamazepine and phenytoin differentially alter extracellular levels of 5-hydroxytryptamine, dopamine and amino acids. Epilepsy Res 2005; 63: 141-149
    CrossrefPubMedGoogle Scholar
  • 53 Bagdy G, Kecskemeti V, Riba P et al. Serotonin and epilepsy. J Neurochem 2007; 100: 857-873
    CrossrefPubMedGoogle Scholar
  • 54 Jithendra C, TEGK Murthy, Upadyay L. Protective role of curcumin in maximal electroshock induced seizures, memory impairment and neurotransmitters in rat brain. Journal of Pre-Clinical and Clinical Research 2008; 2: 35-39
    PubMedGoogle Scholar
  • 55 Seon AL, Seong SH, Xiang HH et al. Piperine from the Fruits of Piper longum with Inhibitory Effect on Monoamine Oxidase and Antidepressant-Like Activity. Chem Pharm Bull 2005; 53: 832-835
    CrossrefPubMedGoogle Scholar
  • 56 Jokeit H, Krämer G, Ebner A. Do antiepileptic drugs accelerate forgetting?. Epilepsy Behav 2005; 6: 430-432
    CrossrefPubMedGoogle Scholar
  • 57 Sudha K, Rao AV, Rao A. Oxidative stress and antioxidants in epilepsy. Clinical Chim Acta 2001; 303: 19-24
    PubMedGoogle Scholar
  • 58 Zhang G, Asgeirsdóttir HN, Cohen SJ et al. Stimulation of serotonin 2A receptors facilitates consolidation and extinction of fear memory in C57BL/6J mice. Neuropharmacology 2013; 64: 403-413
    CrossrefPubMedGoogle Scholar
  • 59 Buhot MC, Martin S, Segu L. Role of serotonin in memory impairment. Ann Med 2000; 32: 210-221
    CrossrefPubMedGoogle Scholar
  • 60 Löscher W, Nolting B. The role of technical, biological and pharmacological factors in the laboratory evaluation of anticonvulsant drugs. IV. Protective indices. Epilepsy Res 1991; 9: 1-10
    CrossrefPubMedGoogle Scholar
  • 61 Zadrozniak A, Wojda E, Wlaź A et al. Characterization of acute adverse-effect profiles of selected antiepileptic drugs in the grip-strength test in mice. Pharmacol Rep 2009; 61: 737-742
    PubMedGoogle Scholar
  • 62 Luszczki JJ, Czuczwar SJ. Biphasic characteristic of interactions between stiripentol and carbamazepine in the mouse maximal electroshock-induced seizure model: a three dimensional isobolographic analysis. Naunyn-Schmiedebergs Arch Pharmacol 2006; 374: 51-64
    PubMedGoogle Scholar
  • 63 Tallarida RJ. Drug Synergism and Dose-Effect Data Analysis. Boca Raton, Florida: Chapman & Hall/CRC; 2000: 65-66
    CrossrefGoogle Scholar