GENOTOXIC AND MUTAGENIC STUDIES OF THE ANTIEPILEPTIC DRUG LEVETIRACETAM IN PREGNANT RATS AND THEIR FETUSES
Keywords:
Mutagenicity, Nil, HRAS gene mutation, Comet assay, LevetiracetamAbstract
Objective: Levetiracetam (LEV) is an anti-epileptic drug, initially approved as an adjunct therapy in adult patients with partial-onset seizures, and used as monotherapy treatment during pregnancy. However, very few, if none, investigations have been focused on LEV neurotoxicity or hepatotoxicity at the molecular level. This study aimed to evaluate the genotoxic and mutagenic potential of LEV, in liver and brain tissues of treated pregnant rats and their fetuses during pregnancy.
Methods: LEV was administered to pregnant female albino rats at doses 300 or 600 mg/kg b. w, from gestation days 5-18. Comet assay, DNA fragmentation were performed for detection of DNA damage. Single-stranded conformation polymorphism (SSCP) followed by DNA sequencing were accomplished for detecting possible mutagenicity.
Results: Administration of the two tested doses of LEV resulted in a significant increase of DNA damage as detected by alkaline Comet assay, and an appearance of both apoptotic laddered and smeared DNA in the tissues tested. Moreover, a significant incidence of mutations in exon 2 and 3 of Harvey rat sarcoma viral oncogene (HRAS) gene, were detected in fetal liver and brain tissues respectively, using single-stranded conformation polymorphism (SSCP) and were conï¬rmed by DNA sequencing.
Conclusion: Maternal and fetal DNA damage induced by LEV was evidenced in our study, even at the commonly used therapeutic dose (300 mg/kg), and thus these side effects should be considered when using LEV for long-term during pregnancy.
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Sridharan R. Epidemiology of epilepsy. Curr Sci 2002;6:664-70.
Ozyurek H, Bozkurt A, Bilge S, Ciftcioglu E, Ilkaya F, Bas DB. Effect of prenatal LEV exposure on the motor and cognitive functions of rat offspring. Brain Dev 2010;32:396-403.
Evan G, Kimford JM. Antiepileptic drugs in women with epilepsy during pregnancy. Ann Neurol 2013;74:223-31.
Lyseng-Williamson KA. Levetiracetam: a review of its use in epilepsy. Drugs 2011;71:489-514.
Ikonomidou C, Bittigau P, Koch C, Genz K, Hoester F, Felderhoff-Mueser U, et al. Neurotransmitters and apoptosis in the developing brain. Biochem Pharmacol 2001;62:401-5.
Webb SJ, Monk CS, Nelson CA. Mechanisms of postnatal neurobiological development: implications for human development. Dev Neuropsychol 2001;19:147-72.
Tan TC, de Boer BW, Mitchell A, Delriviere L, Adams LA, Jeffry GP, et al. LEV as a possible cause of fulminant liver failure. Neurology 2008;71:685-6.
Skopp G, Schmitt HP, Pedal I. Fulminant liver failure in a patient on carbamazepine and LEV associated with status epilepticus. Archiv Für Kriminologie 2006;217:161-75.
Brockmoller J, Thomsen T, Wittstock M, Coupez R, Lochs H, Roots I Pharmacokinetics of LEV in patients with moderate to severe liver cirrhosis (child-pugh classes A, B, and C): characterization by dynamic liver function tests. Clin Pharmacol Ther 2005;77:529-41.
Ohman I, Vitols S, Tomson T. Lamotrigine in pregnancy: pharmacokinetics during delivery, in the neonatal, and during lactation. Epilepsia 2000;41:709-13.
Chen YH, Chiou HY, Lin HC Lin HL. Effect of seizures during gestation on pregnancy outcomes in women with epilepsy. Arch Neurol 2009;66:979-84.
Alberto F, Eugenio S. Ras in cancer and developmental diseases. Genes Cancer 2011;2:344–58.
Yoko A, Tetsuya N, Hiroshi K, Kenji K, Hirofumi O, Yukichi T, et al. Germline mutations in HRAS proto-oncogene cause Costello syndrome. Nat Genet 2005;37:1038–40.
NRC. Guide for the Care and Use of Laboratory Animals. National Research Council. National Academy Press: Washington, DC; 1996.
Snell GD. Biology of the laboratory mouse. Dover Publications: New York; 1956. p. 1-497.
Mcclain RM, Becker BA. Teratogenicity, fetal toxicity and placental transfer of lead nitrate in rats. Toxicol Appl Pharmacal 1975;31:72-82.
Tice RR, gurell EA, Anderson V, Burlinson B, Hartmann A, Kobayashi H, et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 2000;35:206–21.
Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. CSH Cold Spring Harbor Press, NY; 1989.
Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988;16:1215.
Donnelly PJ, Devereux TR, Foley JF, Maronpot RR, Anderson MW, Massey TE. Activation of K-ras in aflatoxin B1-induced lung tumors from mice. Carcinogenesis 1996;17:1735-40.
Phulen Sarma, Anusuya Bhattacharyya. models of epilepsy used in antiepileptic drug discovery: a review. Int J Pharm Pharm Sci 2014;6:975-1491.
Aleksandra F, Ranko S, Snjezana M, Davor Z, Darko M, Romana G, et al. Transplacental genotoxicity of antiepileptic drugs: Animal model and pilot study on mother/newborn cohort. Reprod Toxicol 2010;30:613-8.
Cohen GM, Sun X, Snowden RT, Dinsdale D, Skilleter DN. Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation. Biochemical J 1992;286:331-4.
Olney JW, Jevtovic-Todorovic V, Ikonomidou C. Do pediatric drugs cause developing neurons to commit suicide? Trends Pharmacol Sci 2004;25:135-9.
Manent JB, Jorquera I, Mazzucchelli I, Depaulis A, Perucca E, Ben-Ari Y, et al. Fetal exposure to GABA-acting antiepileptic drugs generates hippocampal and cortical dysplasias. Epilepsia 2007;48:684-93.
Katz I, Kim J, Gale KN, Kondratyev AD. Effects of lamotrigine alone and in combination with MK-801, phenobarbital, or phenytoin on cell death in the neonatal rat brain. J Pharmacol Exp Ther 2007;322:494-500.
Kim J, Kondratyev A, Gale K. Antiepileptic drug-induced neuronal cell death in the immature brain: effects of carbamazepine, topiramate and levetiracetam as monotherapy vs. polytherapy. J Pharmacol Exp Ther 2007;323:165-73.
Kwan P, Brodie MJ. The potential role of drug transporters in the pathogenesis of medically intractable epilepsy. Epilepsia 2005;46:224-35.
Aasim A, Christopher DA. Acute liver failure following levetiracetam therapy for seizure prophylaxis in traumatic brain injury. Case Reports Clin Med 2012;1:41-4.
Huda AM, Muna AM. Chronic histopathological effects of levetiracetam on some internal organs of adult albino rats. Egyptian J Forensic Sci 2015;5:41-5.
Ozden H, Kabay SC, Toker A, Ustüner MC, Ozbayer C, Ustüner D, et al. The effects of levetiracetam on urinary 15f-2t-isoprostane levels in epileptic patients. Seizure 2010;19:514-6.
Meshkibaf MH, Miladpoor B, SholeVar F, Abdollahi A. Chronic effect of gabapentin on liver function in the adult male rat. Acta Med Iran 2013;51:830-3.
Newsome SD, Xue LY, Jennings T, Castaneda GY. Levetiracetam-induced diffuse interstitial lung disease. J Child Neurol 2007;22:628-30.
Patrycja K, Erik T, Ewa G. Effects of valproic acid and levetiracetam on viability and cell cycle regulatory genes expression in the OVCAR-3 cell line. Pharmacol Rep 2012;64:157-65.
Simon D, Fish D. The treatment of epilepsy; 2004. p. 443-50.
Genton P, Van Vleymen B. Piracetam and levetiracetam: close structural similarities but different pharmacological and clinical profiles. Epileptic Disorders 2000;2:99-105.
Hunt S, Craig J, Russell A, Guthrie E, Parsons L, Robertson I, et al. Levetiracetam in pregnancy: preliminary experience from the UK epilepsy and pregnancy register. Neurology 2006;67:1876-9.
Sengul T, Akin T, Mehmet E, Nevin K, Hamit O, Nurten K. Genotoxic effects of prenatal exposure to levetiracetam during pregnancy on rat offspring. In Vivo 2015;29:77-82.
Physician’s desk reference (PDR). 55th ed. Montvale NJ. Medical Economics; 2001. p. 3206-9.
Nina I, Ofer S, Meir B, Jing Z, Michelle M, Boris Y, et al. the Developmental outcome of levetiracetam, its major metabolite in humans, 2 pyrrolidinones N-butyric acid, and Its enantiomer (R)-α-ethyl-oxo-pyrrolidine acetamide in a mouse model of teratogenicity. Epilepsia 2003;44:1280–8.
Patsalos PN. Pharmacokinetic profile of levetiracetam: toward ideal characteristics. Pharmacol Ther 2000;85:77–85.
Yang H, Liu C, Yang D, Zhanga H, Xia Z. Comparative study of cytotoxicity, oxidative stress and genotoxicity induced by four typical nanomaterials: the role of particle size, shape, and composition. J Appl Toxicol 2008;29:69-78.
Bjelland, Seeberg E. Mutagenicity, toxicity, and repair of DNA base damage induced by oxidation. Mutat Res Fundam Mol Mech Mutagen 2003;531:37-80.
Atkinson DE, Boyd RD, Sibley CP. Placental transfer. In: Neill JD. Ed. Knobil and Neill’s Physiology of Reproduction. Elsevier; 2006. p. 2787-846.