Bumetanid’in HT-22 Hipokampal Hücre Hattında Pentilentetrazol Kaynaklı Hücresel Hasar ve Oksidatif Stres Üzerine Etkisinin Araştırılması

Muzaffer Türkoğlu; Ali Tokgöz; Songül Bavli; Sebahattin Karabulut; Ayşegül Öztürk

EN TR

The Investigation of the Effect of Bumetanide on Pentylenetetrazol-Induced Cellular Damage and Oxidative Stress in the HT-22 Hippocampal Cell Line

Abstract

The present study aimed to investigate the potential neuroprotective effect of bumetanide (BMT), a Na⁺-K⁺-2Cl⁻ cotransporter-1 (NKCC1) inhibitor, against pentylenetetrazol (PTZ)-induced epileptiform cellular damage and to evaluate its association with oxidative stress parameters in the HT-22 neuronal cell line. HT-22 cells were divided into four groups: control, PTZ (60 mM), BMT, and BMT + PTZ. Cells were pretreated with BMT at different concentrations (1, 2, 4, and 8 µM) prior to exposure to PTZ. Cell viability was assessed using the XTT assay. Total antioxidant status (TAS) and total oxidant status (TOS) were measured to evaluate cellular oxidative stress. Statistical analyses were performed using one-way ANOVA followed by Tukey’s post-hoc test. PTZ exposure significantly reduced cell viability and induced oxidative stress in HT-22 cells. BMT pretreatment, particularly at the 2 µM concentration, significantly attenuated PTZ-induced cell death (p < 0.05). At this dose, BMT increased TAS levels while significantly limiting the PTZ-induced elevation in TOS. BMT administered alone did not significantly affect cell viability. These findings indicate that BMT exerts a dose-dependent protective effect against PTZ-induced neuronal damage. This study demonstrates that bumetanide pretreatment provides neuroprotection against PTZ-induced cellular injury in HT-22 neurons. The observed protective effect appears to be mediated not by direct antioxidant activity, but rather through modulation of chloride homeostasis, leading to reduced neuronal hyperexcitability and secondary suppression of oxidative stress responses. Targeting NKCC1-dependent upstream mechanisms may represent a complementary therapeutic strategy for epileptiform neuronal damage.

Keywords

Bumetanid’in HT-22 Hipokampal Hücre Hattında Pentilentetrazol Kaynaklı Hücresel Hasar ve Oksidatif Stres Üzerine Etkisinin Araştırılması

Öz

Bu çalışmanın amacı, Na⁺-K⁺-2Cl⁻ kotransporter-1 (NKCC1) inhibitörü olan bumetanidin (BMT), pentilentetrazol (PTZ) ile indüklenen epileptiform hücresel hasara karşı olası nöroprotektif etkisini ve bu etkinin oksidatif stres parametreleri ile ilişkisini HT-22 nöronal hücre hattında araştırmaktır. HT-22 hücreleri kontrol, PTZ (60 mM), BMT ve BMT + PTZ olmak üzere dört gruba ayrılmıştır. Hücreler, farklı konsantrasyonlarda (1, 2, 4 ve 8 µM) BMT ile ön işleme tabi tutulduktan sonra 60 mM PTZ’ye maruz bırakılmıştır. Hücre canlılığı XTT testi ile değerlendirilmiştir. Hücresel oksidatif durumun belirlenmesi amacıyla total antioksidan seviye (TAS) ve total oksidan seviye (TOS) ölçümleri gerçekleştirilmiştir. Veriler tek yönlü ANOVA ve Tukey post-hoc testi kullanılarak analiz edilmiştir. PTZ uygulaması, HT-22 hücrelerinde hücre canlılığını anlamlı düzeyde azaltmış ve oksidatif stres belirteçlerinde belirgin artışa neden olmuştur. BMT ön tedavisinin özellikle 2 µM dozunda, PTZ’ye bağlı hücre ölümünü anlamlı şekilde azalttığı saptanmıştır (p<0.05). Bu dozda BMT uygulaması, TAS düzeylerini artırırken PTZ’ye bağlı TOS artışını anlamlı olarak sınırlamıştır. Tek başına BMT uygulamasının hücre canlılığı üzerinde anlamlı bir etkisi gözlenmemiştir. Bu çalışma, bumetanid ön tedavisinin PTZ ile indüklenen HT-22 hücre hasarına karşı nöroprotektif etki gösterdiğini ortaya koymaktadır. Bu etkinin, doğrudan bir antioksidan mekanizmadan ziyade, klorür homeostazının düzenlenmesi yoluyla nöronal hipereksitabilitenin azaltılmasına ve buna bağlı sekonder oksidatif stres yanıtlarının baskılanmasına bağlı olduğu düşünülmektedir. Bulgularımız, NKCC1 inhibisyonunun epileptiform hasarın upstream patofizyolojik mekanizmalarını hedefleyen tamamlayıcı bir terapötik yaklaşım olabileceğini desteklemektedir.

Anahtar Kelimeler

Destekleyen Kurum

TÜBİTAK

Proje Numarası

1919B012305670

Etik Beyan

Bu çalışma sekonder hücre hattında (HT-22 Mouse Hippocampal Neuronal Cell Line) gerçekleştirildiği için Etik Kurul izin belgesi gerekmemektedir.

Teşekkür

Bu çalışma, TÜBİTAK Bilim İnsanı Destek Programları Başkanlığı (BİDEB) tarafından yürütülen, 2209-A Üniversite Öğrencileri Araştırma Projeleri Destekleme Programı 2023 yılı 1. dönem kapsamında 1919B012305670 başvuru numarasıyla desteklenmiştir.

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Ayrıntılar

Birincil Dil

Türkçe

Konular

Klinik Tıp Bilimleri (Diğer)

Bölüm

Araştırma Makalesi

Yazarlar

Muzaffer Türkoğlu
0009-0000-0085-2332
Türkiye

Ali Tokgöz
0009-0000-3752-9483
Türkiye

Songül Bavli
0000-0002-1688-7828
Türkiye

Sebahattin Karabulut ^*
0000-0002-3261-4125
Türkiye

Ayşegül Öztürk
0000-0001-8130-7968
Türkiye

Yayımlanma Tarihi

14 Nisan 2026

Gönderilme Tarihi

26 Ocak 2026

Kabul Tarihi

23 Mart 2026

Yayımlandığı Sayı

Yıl 2026 Cilt: 4

IZ

https://izlik.org/JA64DD28EP

Kaynak Göster

RIS / Bibtex

APA

Türkoğlu, M., Tokgöz, A., Bavli, S., Karabulut, S., & Öztürk, A. (2026). Bumetanid’in HT-22 Hipokampal Hücre Hattında Pentilentetrazol Kaynaklı Hücresel Hasar ve Oksidatif Stres Üzerine Etkisinin Araştırılması. Health Services Research Journal, 4. https://izlik.org/JA64DD28EP