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

Öz

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.

Anahtar Kelimeler

• Valproic Acid
• Epilepsy
• Pentylenetetrazole
• DNA Damage
• Apoptosis

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

Öz

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.

Anahtar Kelimeler

• Valproik Asit
• Epilepsi
• Pentilentetrazol
• DNA Hasarı
• Apoptozis

Teşekkür

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

Kaynakça

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