Erkan TUNCAY, Yusuf OLĞAR, Belma TURAN, Sinan DEĞİRMENCİ, Ayşegül DURAK, Naci ERTÜRK, Muhammet Talha AKBAŞ, Mehmet Cihangir DENİZ, Muhammed Furkan ERCİYAS, Burak Tahir YAZAR, Mustafa Salih YILMAZ, Ahmet AYGÜN

SGLT2 İnhibitörü Dapagliflozinin Hiperglisemi-Aracılı Kalp Fonksiyon Bozukluğu Üzerindeki Etkisinin Moleküler Temellerinin İncelenmesi

Amaç: Sodyum glukoz ko-transporter (SGLT) inhibitörlerinin diyabetli bireylerde glukoz re-absorbsiyonunu inhibe ederek kan şekerini düşürdüğü ileri sürülmektedir. Bu inhibitörlerin kalp fonksiyonu üzerindeki etkileri henüz tam olarak bilinmemesine karşın, Na+/H+-değiştokuşçusu (NHE) inhibitörü gibi etkiler gösterebildiği ileri sürülmektedir. Bu çalışmada, SGLT2-inhibitörü olarak Dapagliflozin (DAPA) in vitro olarak sıçan embriyonik hiperglisemik ventriküler hücrelerinde (H9c2-hücre hattı), hücre seviyesindeki etki mekanizmasının elektrofizyolojik yöntemler kullanılarak incelenmesi hedeflenmiştir. Gereç ve Yöntem: H9c2 hücreleri deney koşullarına hazır hale getirildikten sonra, H9c2’ler bir grubu 25 mM glukoz ile 24-saat veya 48-saat 37 °C’de inkübe edilerek hiperglisemik kardiyomiyositler (HG) elde edilmiş, bir diğer grubun inkübasyonu ise 25 mM glukoz ve DAPA (D185360, TORONTO Research Chemicals; 100 nM veya 1 µM) ile aynı sürelerde gerçekleştirilmiştir. Hücreiçi iyon derişimleri ([X]in), [ROS]in ve mitokondri membran potansiyeli (MMP) ölçümleri hücreler özel floresans boyalarla yüklenerek (DCFDA, FluoZin-3AM, JC-1, SNARF-1AM, FURA-2AM ve SBFI) konfokal mikroskobu ve mikrospektrofluorometre kullanılarak ölçülmüştür. Gruplar arası karşılaştırmalar student t-testi ile gerçekleştirilmiş ve anlamlılık değeri 0,05’den küçük değerler kabul edilmiştir. Bulgular: Normal ve hiperglisemik hücrelerde 1 µM DAPA uygulamasında toksik etkiler gözlendiğinden, tüm incelemeler 100 nM DAPA uygulaması ile yapılmıştır. [Na+] in değeri bu uygulamalarda değişmezken, HG inkübasyonda (48-saat) istatistiksel olarak anlamlı düzeyde artış olan [H+] in değeri, DAPA uygulaması ile (24-saat ve 48-saat) çok belirgin olarak artmıştır. Buna karşın, HG’li hücrelerde (24-saat ve 48-saat) artmış olan [Zn+2] in değerlerini DAPA uygulaması etkilememiştir. Buna karşın, HG’li hücrelerde (24-saat ve 48-saat) artmış olan [Ca+2] in değerlerinin DAPA uygulaması ile normal değerlere yakın olduğu gözlenmiştir. Bunlara ek olarak, HG koşullarında önemli derecelerde artmış olan [ROS]in değerinin DAPA ile normal değerler civarında gözlendiği, fakat HG sonucu depolarize durumdaki MMP’lerini etkilemediği sonuçları elde edilmiştir. Sonuç: Sonuç olarak elde edilen verilerimiz, SGLT2 inhibitörlerinin HG hücrelerde, artan [H+] in ve [ROS]in baskılayarak, artmış olan bazal [Ca+2] in’nin düzelmesine ve böylece kalbin kasılma-gevşeme aktivitesinin HG koşullarında normal fonksiyon göstermesine yol açabileceğini işaret etmektedir.

An Investigation on Molecular Basis of the Effects of SGLT2 İnhibitor Dapagliflozin on Hyperglycemia-Associated Heart Dysfunction

Objectives: It has been reported that sodium glucose co-transporter (SGLT) inhibitors lowering blood glucose in diabetic patients via inhibiting glucose re-absorption. Although it has not been clearly identified the effects of these inhibitors on heart function, these inhibitors may act like Na+/ H+-exchanger (NHE) inhibitors. In this study, we were aimed to clarify the underlying cellular mechanism of a SGLT2-inhibitor, Dapagliflozin (DAPA), on hyperglycemic embryonic rat ventricular cells (H9c2 cell line) via electrophysiological measurements. Materials and Methods: One group of H9c2 cells were incubated with high glucose (25 mM) medium for 24-hours and 48-hours at 37 °C to obtain hyperglycemic (HG) cardiomyocytes. Another group of H9c2 cells were incubated together with high glucose medium and DAPA (D185360, TORONTO Research Chemicals; 100 nM or 1 µM). Intracellular ion concentrations, ([X]in), reactive oxygen species, ([ROS]in) and mitochondria membrane potential, (MMP) were monitored via specific fluorescent dyes (DCFDA, FluoZinc, JC-1, SNARF, FURA-2AM and SBFI) with confocal microscope and microspectrofluorometer. All data are presented as mean (± SEM) and statistical analysis performed by student t-test. Significance level considered at p

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