Pregabalinin Glutamat Eksitotoksisitesine Karşı Nöroprotektif Etkinliğinde Oksidatif ve Nitrozatif Yolların Rolü

Abstract

Amaç: Bu çalışma pregabalinin nitrik oksit ve oksidatif stres yolakları üzerinden glutamat ile indüklenen sitotoksisite üzerindeki nöroprotektif etkisini araştırmak için tasarlandı. Gereç ve Yöntem: Deneyde C6 glioma hücre hattı kullanılmıştır. Pregabalinin glutamatın neden olduğu sitotoksisiteden sonra glial hücre ölümü üzerindeki etkisini değerlendirmek için dört grup oluşturuldu. Kontrol grubu herhangi bir işlem görmedi. Glutamat grubundaki hücreler 24 saat 10 mM glutamat ile muamele edildi. Pregabalin grubundaki hücreler, 24 saat boyunca çeşitli konsantrasyonlarda (3,75; 7,5; 15; 30 ve 60µM) pregabalin ile muamele edildi. Pregabalin+glutamat grubundaki hücreler, 1 saat boyunca çeşitli konsantrasyonlarda (3,75; 7,5; 15; 30 ve 60µM) pregabalin ile ön işleme tabi tutuldu ve ardından 24 saat boyunca 10mM glutamata maruz bırakıldı. Hücre canlılığı, XTT tahlili ile değerlendirildi. Hücrelerdeki toplam antioksidan ve oksidan seviyesi (TAS-TOS), indüklenebilir nitrik oksit (iNOS), nitrik oksit (NO) ve nöronal nitrik oksit (nNOS) seviyeleri ticari kitlerle ölçüldü. Bulgular: 30 ve 60 µM konsantrasyondaki pregabalin, glutamatın neden olduğu sitotoksisiteden sonra C6 hücrelerinde hücre canlılığını önemli ölçüde arttırdı (p<0,01). Pregabalin (30µM)+glutamat, tedavi edilmemiş glutamat grubuna (p<0,05) kıyasla C6 hücrelerinde TOS seviyelerini önemli ölçüde azaltırken, TAS seviyesini artırmıştır (p<0,05). Ayrıca pregabalin, glutamatın indüklediği sitotoksisite sonrasında C6 hücrelerinde iNOS ve NO seviyesini azaltırken (p>0,05) nNOS düzeyini değiştirmemiştir (p>0,05). Sonuç: Pregabalin, sitotoksisite üzerinde koruyucu ve antioksidan özelliklere sahiptir. Eğer in vivo ve vitro daha fazla araştırma bu iddiayı desteklerse Alzheimer ve Parkinson gibi nörodejeneratif hastalıklar için etkili bir takviye olabilir.

Keywords

• Glutamat Eksitotoksisitesi
• Pregabalin
• C6 Glioma
• Oksidatif Stres
• Nitrozatif Stres

The Role of Oxidative and Nitrosative Pathways in the Neuroprotective Effect of Pregabalin Against Glutamate Excitotoxicity

Abstract

Introduction: This study was designed to investigate the neuroprotective effect of pregabalin on glutamate-induced cytotoxicity via nitric oxide and oxidative stress pathways. Material Method: C6 glioma cell line was used in the experiment. Four groups were created to evaluate the effect of pregabalin on glial cell death after glutamate-induced cytotoxicity. Control group did not receive any treatment. Cells in the glutamate group were treated with 10 mM glutamate for 24 hours. Cells in the pregabalin group were treated with pregabalin at various concentrations (3.75, 7.5, 15, 30, and 60 µM) for 24 hours. Cells in the pregabalin+glutamate group were pretreated with pregabalin at various concentrations (3.75, 7.5, 15, 30, and 60 µM) for 1 h and then exposed to 10 mM glutamate for 24 h. Cell viability was assessed by XTT assay. Total antioxidant and oxidant levels (TAS-TOS), inducible nitric oxide (iNOS), nitric oxide (NO), and neuronal nitric oxide (nNOS) levels in cells were measured with commercial kits. Results: Pregabalin at 30 and 60 µM concentrations significantly increased cell viability in C6 cells after glutamate-induced cytotoxicity (p<0,01). Pregabalin (30µM)+glutamate significantly decreased TOS levels in C6 cells compared to the untreated glutamate group (p<0,05), while it increased TAS level (p<0,05). In addition, pregabalin decreased iNOS and NO levels in C6 cells after glutamate-induced cytotoxicity (p>0,05) but did not change nNOS levels (p>0,05). Conclusion: Pregabalin has protective and antioxidant properties on cytotoxicity. If further in vivo and in vitro research supports this claim, it may be an effective supplement for neurodegenerative diseases such as Alzheimer's and Parkinson's.

Keywords

• Glutamate Excitotoxicity
• Pregabalin
• C6 Glioma
• Oxidative Stress
• Nitrosative Stress

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