Hidrokortizol ve B12 Vitamini VIP ve GAL Düzeylerini Değiştirerek SHSY-5Y Hücrelerini Glutamat Eksotoksisitesine Karşı Korur

Yıl 2024, Cilt: 9 Sayı: 4, 754 - 761, 31.12.2024

Sıdıka Genç , Kübra Karabulut , Esmanur Niğde , Ali Taghizadehghalehjoughi

https://doi.org/10.35229/jaes.1564954

Öz

Giriş: Hücre dışı glutamat seviyelerinin uzun süreli yükselmesi hücre içi olayları tetikler, glutamat ekzositozunu artırır ve apoptotik yolları aktive ederek Alzheimer hastalığına neden olur. Literatürde B12 vitamininin çeşitli hastalıklarda anti-inflamatuar ve anti-apoptotik aktiviteler gösterdiği bildirilmiştir. Hidrokortizon tedavisi ayrıca mikroglia ve astrosit hiperaktivasyonunu önemli ölçüde inhibe ederek pro-inflamatuar sitokinleri en aza indirir ve nöroinflamasyonu azaltır. Bu nedenle, çalışmamız, hidrokortizol ve B12 kombinasyonunun in vitro Alzheimer modelinde oksidatif stres ve vazoaktif intestinal peptit ve galanin seviyeleri üzerindeki terapötik etkilerini değerlendirmeyi amaçlamıştır.
Yöntem: Alzheimer modelini oluşturmak için nöroblastoma hücre hattı (SH-SY5Y) kültürlendi. Daha sonra, kontrol grubu hariç tüm hücreler, ekzotoksisite oluşturmak için Glutamat (10-5 mM) uygulanarak 24 saat boyunca farklı dozlarda HC ve B12 kombinasyonu ile tedavi edildi. Sonuçlar MTT ve ELISA testleri kullanılarak değerlendirildi.
Sonuçlar: Sonuçlar incelendiğinde, istisnai olarak yüksek doz kombinasyon gruplarının glutamat ekzotoksisitesine karşı koruyucu aktivite gösterdiği belirlendi. HC+B12 25 µg/ml grupları istatistiksel olarak en anlamlı sonuç olarak gözlemledi. Sonuçlarımıza göre, HC+B12 25 µg/ml grubunda oksidatif stres azaldı ve hücre canlılığı arttı. Elde edilen diğer analizlerle korelasyon içinde vazoaktif intestinal peptit ve galanin seviyelerinde de önemli değişiklikler gözlendi.
Sonuç: Bu çalışma, birincil nöronlarda oksidatif stresi ve glutamat eksitotoksisitesini önlemek için hidrokortizonla kombine B12 vitamininin potansiyelini bildiren ilk çalışmadır. Nörodejeneratif hastalıklarda klinik uygulamasının daha fazla araştırılması için bir temel sağlar.

Anahtar Kelimeler

Glutamat, VIP, GAL, eksitotoksisite

Hydrocortisone and vitamin B12 protect SHSY-5Y cells against glutamate excitotoxicity by altering VIP and GAL levels

Öz

Background: Prolonged elevation of extracellular glutamate levels triggers intracellular events, increases glutamate excitotoxicity, and activates apoptotic pathways, causing Alzheimer's disease (AD). The literature has reported that vitamin B12 exhibits anti-inflammatory and anti-apoptotic activities in various diseases. Hydrocortisone (HC) therapy also substantially inhibits microglia and astrocyte hyperactivation, minimizing pro-inflammatory cytokines and reducing neuroinflammation. That is why our study aimed to evaluate the therapeutic effects of HC and B12 combination on oxidative stress and VIP and GAL levels in an in vitro Alzheimer's model.
Method: To create the Alzheimer's model, the neuroblastoma cell line (SH-SY5Y) was cultured. Then, all cells except the control group were treated with different doses of HC and B12 combination for 24 hours by applying Glutamate (10-5 mM) to create excitotoxicity. The results were evaluated using MTT and ELISA tests.
Results: When the results were examined, it was determined that exceptionally high-dose combination groups showed protective activity against glutamate excitotoxicity. HC+B12 25 µg/ml groups observed the most statistically significant results. According to our results, oxidative stress decreased in the HC+B12 25 µg/ml group, and cell viability increased. Significant changes were also observed in Vasoactive Peptide (VIP) and Galanin (GAL) levels in correlation with other analyses obtained.
Conclusion: This study is the first to report the potential of vitamin B12 combined with hydrocortisone to prevent oxidative stress and glutamate excitotoxicity in primary neurons. It provides a basis for further investigating its clinical application in neurodegenerative diseases.

Anahtar Kelimeler

Glutamate, VIP, GAL, excitotoxicity.

Kaynakça

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