Valproik Asit’in Sıçanlarda Pentilentetrazol ile Oluşturulan Epileptik Nöbet Sonrası Hipokampüs ve Kortekste Meydana Gelen DNA Hasarı ve Apoptozis Üzerine Etkisi

Year 2020, , 2924 - 2931, 15.12.2020

Mahmut Ekici Ahmet Şevki Taşkıran

https://doi.org/10.21597/jist.730381

Cited By: 1

Abstract

Valproik asit (VPA) epilepsi tedavisinde yaygın olarak kullanılan bir kimyasal ajandır. Son zamanlarda yapılan çalışmalar VPA’in nöronlar üzerine bazı olumsuz etkilerinin olabileceğini göstermiştir fakat bu konu henüz netlik kazanmamıştır. Bu çalışmanın amacı VPA’in sıçanlarda pentilentetrazol (PTZ) ile oluşturulan epileptik nöbet sonrası hipokampüs ve kortekste meydana gelen DNA hasarı ve apoptozis üzerine etkisini ortaya koymaktır. Çalışmada 18 adet 230-250 gram ağırlığında erkek sıçan kullanılmıştır. Sıçanlar kontrol (serum fizyolojik 1 ml kg-1+ serum fizyolojik 1 ml kg-1; n=6), PTZ (serum fizyolojik 1 ml kg-1 + PTZ; n=6) ve VPA (150 mg kg-1 VPA + PTZ; n=6) olmak üzere üç gruba ayrılmıştır. PTZ grubu ve VPA grubuna belirtilen dozlarda uygulanmasından yarım saat sonra 45 mg kg-1 pentilentetrazol intraperitoneal olarak uygulanarak nöbet oluşumu sağlanmıştır. Nöbet oluşumundan 24 saat sonra tüm sıçanların beyin dokuları çıkarılmış, korteks ve hipokampüs bölgeleri alınmıştır. Korteks ve hipokampüs dokularında enzim bağlı immüno sorbent (ELISA) yöntemi ile DNA hasar belirteci olan 8-hidroksi-2'-deoksiguanozin (8-OhDG) ve apoptozis belirteci olan caspase-3 ölçülmüştür. One way ANOVA varyans analizi ile istatistiksel değerlendirme yapılmıştır. PTZ grubunda 8-OhDG seviyesi hem korteks hem de hipokampüste kontrol grubuna göre artmıştır (p<0.05). Bununla birlikte, VPA nöbet sonrası 8-OhDG seviyesini korteks ve hipokampüste PTZ grubuna kıyasla yükseltmiştir (p<0.05). Ayrıca, kortekste caspase-3 seviyesi PTZ grubunda kontrole göre anlamlı olarak artmıştır (p<0.05). Bunun yanı sıra, VPA kortekste PTZ grubuna kıyasla caspase-3 seviyesini anlamlı olarak yükseltmiştir (p<0.05). Sonuç olarak, VPA sıçanlarda nöbet sonrası meydana gelen DNA hasarı ve apoptozisi arttırmıştır.

Keywords

Valproik Asit, Epilepsi, Pentilentetrazol, DNA Hasarı, Apoptozis

Thanks

Bu çalışma için finansal destek alınmamıştır. Fakat Sivas Cumhuriyet Üniversitesi Tıp Fakültesi Araştırma Merkezi (CÜTFAM) alt yapısı kullanılarak çalışma gerçekleştirilmiştir.

The Effect of Valproic Acid on DNA Damage and Apoptosis After Pentylenetetrazole-induced Epileptic Seizure Generated in the Hippocampus and Cortex in Rats

Abstract

the treatment of epilepsy. Recent studies have shown that VPA may have some negative effects on nerve cells, but this issue has not been clarified yet. The aim of this study was to investigate the effect of VPA on DNA damage and apoptosis after pentylenetetrazole (PTZ) induced epileptic seizure generated in the hippocampus and cortex in rats. In the study, 18 males 230-250 grams of rats were used. Rats are divided into three groups as control (physiological serum 1 ml kg-1 + physiological serum 1 ml kg-1; n=6), PTZ (physiological serum 1 ml kg-1 + PTZ; n=6) and VPA (150 mg kg-1 VPA+PTZ; n=6). Seizure was induced by administering 45 mg kg-1 pentylenetetrazole intraperitoneally twenty minutes after the administration at the indicated doses to the PTZ group and VPA group. The brain tissues of all rats were removed, and cortex and hippocampus areas were separated 24 hours after seizure. 8-hydroxy-2'-deoxyguanosine (8-OhDG), which is a DNA damage marker, and caspase-3, which is a marker of apoptosis, were measured in the cortex and hippocampus tissues by the enzyme-linked immunosorbent assay (ELISA) method. One-way ANOVA variance analysis was used for statistical evaluation. In the PTZ group, the 8-OhDG level increased in both cortex and hippocampus compared to the control group (p<0.05). Therefore, VPA enhanced the 8-OhDG level after seizure compared to the PTZ group in the cortex and hippocampus (p<0.05). In addition, the level of caspase-3 in the cortex significantly raised compared to the control in the PTZ group (p<0.05). Moreover, VPA significantly improved the level of caspase-3 in the cortex compared to the PTZ group (p<0.05). In conclusion, VPA increased DNA damage and apoptosis after seizures in rats.

Keywords

Valproic Acid, Epilepsy, Pentylenetetrazole, DNA Damage, Apoptosis

References

• Cerna T, Hrabeta J, Eckschlager T, Frei E, Schmeiser HH, Arlt VM, Stiborová M, 2018. The histone deacetylase inhibitor valproic acid exerts a synergistic cytotoxicity with the DNA-damaging drug ellipticine in neuroblastoma cells. International Journal of Molecular Sciences, 19: 164.
• Chateauvieux S, Morceau F, Diederich M, 2014. Valproic Acid. In Encyclopedia of Toxicology: Third Edition, 905-908.
• Chen X, Wang H, Zhou M, Li X, Fang Z, Gao H, Li Y, Hu W, 2018. Valproic acid attenuates traumatic brain injury-induced inflammation in vivo: Involvement of autophagy and the Nrf2/ARE signaling pathway. Frontiers in Molecular Neuroscience, 11: 117.
• Chu T, Zhou H, Lu L, Kong X, Wang T, Pan B, Feng S, 2015. Valproic acid-mediated neuroprotection and neurogenesis after spinal cord injury: From mechanism to clinical potential. In Regenerative Medicine, 10: 193-209.
• Demirel Yılmaz B, Eren B, Sağır D, Eren Z, Başardı Gökçe A, 2019. Stereological examination of curcumin’s effects on hippocampal damage caused by the anti-epileptic drugs phenobarbital and valproic acid in the developing rat brain. Acta Histochemica, 121: 430-436.
• El-Deeb OS, Ghanem HB, El-Esawy RO, Sadek MT, 2019. The modulatory effects of luteolin on cyclic AMP/Ciliary neurotrophic factor signaling pathway in experimentally induced autoimmune encephalomyelitis. IUBMB Life, 71: 1401–1408.
• Ergul M, Bakar Ateş F, 2019. RO3280: A Novel PLK1 Inhibitor, Suppressed the Proliferation of MCF-7 Breast Cancer Cells Through the Induction of Cell Cycle Arrest at G2/M Point. Anti-Cancer Agents in Medicinal Chemistry, 19: 1846-1854.
• Golechha M, Sarangal V, Bhatia J, Chaudhry U, Saluja D, Arya DS, 2014. Naringin ameliorates pentylenetetrazol-induced seizures and associated oxidative stress, inflammation, and cognitive impairment in rats: Possible mechanisms of neuroprotection. Epilepsy and Behavior, 09: 98‐102.
• Haznedar P, Doğan Ö, Albayrak P, Öz Tunçer G, Teber S, Deda G, Eminoglu FT, 2019. Effects of levetiracetam and valproic acid treatment on liver function tests, plasma free carnitine and lipid peroxidation in childhood epilepsies. Epilepsy Research, 153: 7-13.
• Henshall DC, Simon RP, 2005. Epilepsy and apoptosis pathways. In Journal of Cerebral Blood Flow and Metabolism, 25: 1557-1572.
• Kruger NJ, 2009. The Bradford method for protein quantitation. The protein protocols handbook. Humana Press, Totowa, NJ, 17-24.
• Lotfy DM, Safar MM, Mohamed SH, Kenawy SA, 2018. Effect of valproic acid alone or combined with low dose gamma irradiation in modulating PTZ-induced convulsions in rats involving AKT/m-TOR pathway. Life Sciences, 212: 261-266.
• Michoulas A, Tong V, Teng XW, Chang TKH, Abbott FS, Farrell K, 2006. Oxidative stress in children receiving valproic acid. Journal of Pediatrics, 149: 692-696.
• Nanau RM, Neuman MG, 2013. Adverse drug reactions induced by valproic acid. In Clinical Biochemistry, 46: 1323-1338.
• Racine RJ, 1972. Modification of seizure activity by electrical stimulation: I. after-discharge threshold. Electroencephalography and Clinical Neurophysiology, 32: 269-279.
• Shakirullah Ali N, Khan A, Nabi M, 2014. The Prevalence, Incidence and Etiology of Epilepsy. Int J Clin Exp Neurol, 2014: 29-39.
• Shona SI, Rizk AA, El Sadik AO, Emam HY, Ali EN, 2018. Effect of valproic acid administration during pregnancy on postnatal development of cerebellar cortex and the possible protective role of folic acid. Folia Morphologica (Poland), 77: 201-209.
• Suda S, Katsura KI, Kanamaru T, Saito M, Katayama Y, 2013. Valproic acid attenuates ischemia-reperfusion injury in the rat brain through inhibition of oxidative stress and inflammation. European Journal of Pharmacology, 707: 26-31.
• Taskıran AS, Gumus E, Gunes H, Cetindag A, Ozdemir E, 2018. The Protective Effects of Vitamin B 12 on Pentylenetetrazole-Induced Seizures in Rats. Anatomy Physiology & Biochemistry International Journal, 4: 1–5.
• Valavanidis A, Vlachogianni T, Fiotakis C, 2009. 8-Hydroxy-2′ -deoxyguanosine (8-OHdG): A critical biomarker of oxidative stress and carcinogenesis. Journal of Environmental Science and Health - Part C Environmental Carcinogenesis and Ecotoxicology Reviews, 27: 120-139.
• Verrotti A, Scardapane A, Franzoni E, Manco R, Chiarelli F, 2008. Increased oxidative stress in epileptic children treated with valproic acid. Epilepsy Research, 78: 171-177.