mTOR inhibisyonu, arka bacak iskemisi/reperfüzyon zedelenmelerinde apoptozu ve oksidatif stresi düzenler

Year 2023, Volume: 48 Issue: 4, 1322 - 1332, 29.12.2023

Zarife Pire , Demet Sinem Güden , Meryem Temiz Reşitoğlu , Sefika Pınar Şenol , Özden Vezir , Nehir Sucu , Bahar Tunçtan , Kafait U. Malik , Seyhan Şahan Fırat

https://doi.org/10.17826/cumj.1353689

Abstract

Amaç: İskemi/reperfüzyon (İ/R)’nun neden olduğu zedelenmeler, oksidan/antioksidan dengenin bozulması ve apopitoz nedeniyle hedef organlarda ve takip eden uzak organlarda gelişebilecek komplikasyonlar ile ilgili ciddi klinik olayları temsil eder. İ/R zedelenmeleriyle ilgili yapılan son çalışmalarda farklı organlarda gelişebilecek İ/R kaynaklı zedelenmelerde rapamisinin memeli hedefi (mTOR)’un etkileri halen tartışılmaya devam etmektedir. Bu çalışmada arka bacak İ/R zedelenmesinde mTOR inhibisyonunun olası koruyucu etkisi ve bu etkisine aracılık edebilecek antiapoptotik ve antioksidan etki mekanizmalarının araştırılması amaçlandı.
Gereç ve yöntem: Wistar sıçanlar dört gruba ayrıldı. Sıçanlarda arka bacak İ/R zedelenmesini oluşturmak için anestezi altında her iki arka ekstremitelerine turnike uygulanarak 4 saat iskemi ve ardından turnikeler açılarak 4 saat reperfüzyon uygulandı. Reperfüzyondan 1 saat önce rapamisin (1 mg/kg) veya salin (4 mL/kg) periton içine uygulandı. Deneylerin sonunda gastroknemius kası, böbrek ve kanları alındı. Kas ve böbrek dokularında İ/R’de gelişebilecek zedelenmeler ribozomal protein S6 (rpS6), ökaryotik başlatma faktörü 4E-bağlayıcı protein 1 (4EBP1), kaspaz-3, Bcl-2 ile ilişkili X proteini (Bax) ve B-hücreli lenfoma (Bcl)-2’nin protein ekspresyonu ve/veya fosforilasyonu ile NADPH oksidaz seviyesi ve toplam antioksidan kapasitesi ölçülerek değerlendirildi.
Bulgular: İ/R'nin neden olduğu organ zedelemeleri artan rpS6, 4EBP1, kaspaz-3, Bax ve azalan Bcl-2 ekspresyon ve/veya fosforilasyonu ile buna eşlik eden toplam antioksidan kapasitede azalma ve NADPH oksidaz seviyesinde artma ile gösterildi. Bir mTOR inhibitörü olan rapamisin uygulaması, İ/R zedelenmesine aracılık eden etkileri ortadan kaldırdı.
Sonuç: Bulgularımız, mTOR sinyal iletisinin etkinleşmesinin oksidan/antioksidan dengedeki bozulmanın sonucu olarak apopitozun etkinleşmesi aracılığıyla arka bacaklarda İ/R’nin neden olduğu doku ve organ zedelenmelerinde önemli bir rol oynadığını göstermektedir.

Keywords

Apopitoz, iskemi/reperfüzyon zedelenmesi, mTOR, oksidatif stres

Project Number

2018-2-TP2-2984

mTOR inhibition modulates apoptosis and oxidative stress in hindlimb ischemia/reperfusion injury

Abstract

Purpose: Ischemia/reperfusion (I/R)-induced injuries represent serious clinical events regarding profound target organ destructions followed by remote organ complications due to the loss of oxidant/antioxidant balance and apoptosis. Recent studies examining the mammalian target of rapamycin (mTOR) during I/R injury in different organs have remained a matter of debate. The current study aimed to explore further the protective and underlying antiapoptotic and antioxidant mechanisms of mammalian target of rapamycin (mTOR) inhibition in hindlimb (HL) schemia/reperfusion (I/R)injury.
Materials and Methods: Occlusion of bilateral hindlimbs for 4 h with tourniquets was carried out under anesthesia to induce I/R for 4 h in rats. Rapamycin (1 mg/kg) or saline (4 mL/kg) was injected intraperitoneally 1 h before reperfusion. Gastrocnemius muscle, kidney, and blood were collected at the end of the experiments for analysis. Muscle and kidney damages were evaluated by measuring protein expression and/or phosphorylation of eukaryotic initiation factor 4E-binding protein 1 (4EBP1), ribosomal protein S6 (rpS6), B-cell lymphoma 2 (Bcl-2), caspase-3, and Bcl-2-associated X protein (Bax) with NADPH oxidase level and total antioxidant capacity in tissues or sera.
Results: I/R-induced organ damages were demonstrated by enhanced phosphorylation and/or expression of rpS6, 4EBP1, caspase-3, and Bax with a significant reduction in Bcl-2 accompanied by a decreased total antioxidant capacity and increased level of NADPH oxidase. Administration of rapamycin, an inhibitor mTOR, protected against I/R-mediated injuries.
Conclusion: Our findings suggest that the activation of mTOR signaling plays a crucial role in HL I/R-triggered organ damages presumably through the activation of apoptosis as a result of oxidant/antioxidant imbalance.

Keywords

apoptosis, reperfusion injury, mTOR, oxidants, antioxidants

Project Number

2018-2-TP2-2984

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