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

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

doi:10.17826/cumj.1353689

EN TR

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

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

Öz

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.

Anahtar Kelimeler

Apopitoz , iskemi/reperfüzyon zedelenmesi , mTOR , oksidatif stres

Kaynakça

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Ayrıntılar

Birincil Dil

İngilizce

Konular

Klinik Tıp Bilimleri (Diğer)

Bölüm

Araştırma Makalesi

Yazarlar

Zarife Pire
0000-0003-0424-9420
Türkiye

Demet Sinem Güden
0000-0001-5423-3641
Türkiye

Meryem Temiz Reşitoğlu
0000-0002-3326-2440
Türkiye

Sefika Pınar Şenol
0000-0002-3019-9589
Türkiye

Özden Vezir
0000-0001-9948-0515
Türkiye

Nehir Sucu
0000-0002-7469-5883
Türkiye

Bahar Tunçtan
0000-0003-3439-7803
Türkiye

Kafait U. Malik
0000-0001-9157-0356
United States

Seyhan Şahan Fırat ^*
0000-0002-8677-6381
Türkiye

Yayımlanma Tarihi

29 Aralık 2023

Gönderilme Tarihi

5 Ekim 2023

Kabul Tarihi

11 Aralık 2023

Yayımlandığı Sayı

Yıl 2023 Cilt: 48 Sayı: 4

DOI

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

IZ

https://izlik.org/JA34ZT49TY

MLA

Pire, Zarife, vd. “mTOR inhibition modulates apoptosis and oxidative stress in hindlimb ischemia/reperfusion injury”. Cukurova Medical Journal, c. 48, sy 4, Aralık 2023, ss. 1322-3, doi:10.17826/cumj.1353689.

Bu eser Creative Commons Atıf-Gayriticari-Türetilemez 4.0 Uluslararası Lisansı ile lisanslanmıştır.

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

Abstract

Keywords

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

Öz

Anahtar Kelimeler

Kaynakça

Ayrıntılar

Birincil Dil

Konular

Bölüm

Yazarlar

Yayımlanma Tarihi

Gönderilme Tarihi

Kabul Tarihi

Yayımlandığı Sayı

DOI

IZ

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