Endoplazmik retikulum stres belirteçlerinden ATF5 ve fosforile IF2 düzeylerinin nöroblastoma hücrelerinde kainik asit muamelesi sonrası incelenmesi

Amaç: Bu çalışmada, amacımız, kainik asite bağlı eksitotoksisite ve endoplazmik retikulum (ER) stres arasındaki ilişkiyi iki majör endoplazmik retikulum stres markırı olan ATF5 ve fosforile olmuş eIF2a analiz ederek incelemektir. Gereç ve Yöntem: Neuroblastoma hücrelerini 1 mM kainik asit ile 24 saat muamele ettik. ATP ölçümü, kainik asit muamelesi yapılan hücrelerde ya da kontrol hücrelerinde bioluminesans bir yöntem ile yapılmıştır. Total protein, kainik asit muamelesi yapılmış ya da kontrol hücrelerinden izole edilmiş ve ATF5 ve fosforile olmuş eIF2 markırları western blot ile incelenmiştir. Bulgular: Kainik asit ile muamele edilmiş neuroblastoma hücrelerinde ATF5 ve fosforile olmuş eIF2a seviyelerinin kontrole göre değişmediğini ilk kez gösterdik. Kainik asit ile muamele edilmiş hücrelerde ATP seviyesinin düştüğünü gösterdik. Sonuç: 1 mM ve 24 saat süren kainik asit muamelesi, ATF5 ve fosforile olmuş eIF2 ile gösterilebilecek endoplazmik retikulum stresi yaratmak için yeterli olmayabilir. Süre ve konsantrasyon olarak arttırılmış kainik asit muamelesi ya da farklı markırlar gerekmektedir. Eksitotoksisiteye bağlı beyin hastalıklarında ER stres yolaklarını araştırmak, yeni tedavi yolları bulmak ve birden fazla hastalığı aynı anda engellemek adına önemli olacaktır.

The investigation of endoplasmic reticulum stress markers ATF5 and phosphorylated eIF2 after kainic acid treatment in neuroblastoma cells

Purpose: The aim of this study was to investigate therelationship between kainic acid induced excitotoxicity andendoplasmic resticulum (ER) stress by analyzing twomajor ER stress markers such as ATF5 andphosphorylated eIF2 in neuroblastoma cells.Materials and Methods: Neuroblastoma cells weretreated with 1 mM kainic acid for 24 hours. ATPmeasurement was performed in kainic acid-treated andvehicle-treated neuroblastoma cells via ATPbioluminescence assay. Total protein was isolated fromkainic acid-treated and control cells. Via western blotting,the expression levels of ATF5 and phosphorylated eIF2were analyzed.Results: We showed for the first time that as a result ofkainic acid treatment in neuroblastoma cells, the proteinexpression levels of ER stress markers ATF5 andphosphorylated eIF2 did not display any change whencompared to control cells. We also showed that ATP levelswere decreased in kainic acid-treated cells.Conclusion: This study may show that the level of stressthat kainic acid causes at 1 mM for 24 hours inneuroblastoma cells was not adequate to lead to ER stresswhich is measurable by ATF5 and phosphorylated eIF2.Either an increased level of treatment of kainic acid viaincreased duration or concentration is necessary ordifferent markers should be tried. The investigation of theER stress pathways in the excitotoxicity-related braindiseases will pave the way for new therapies based on ERstress and combat more than one disease simultaneously

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