Effect of glucagon-like peptide-1 on differentiation of adipose derived mesenchymal stem cells into cardiomyocytes

Özgür Tekin; Yiğit Uyanıkgil; Dilek Taşkıran

doi:10.19161/etd.1180666

EN TR

Effect of glucagon-like peptide-1 on differentiation of adipose derived mesenchymal stem cells into cardiomyocytes

Abstract

Aim: Mesenchymal stem cells can easily differentiate into cardiomyocytes in vitro conditions using various protocols. However, the agents used in these protocols have been reported to have some adverse effects on cell viability. Azacitidine is used to differentiate mesenchymal stem cells into cardiac muscle cells. The aim of the present study was to investigate the effects of Exenatide a GLP-1 receptor agonist, on differentiation and viability of human adipose tissue derived stem cells into cardiomyocytes.

Materials and Methods: The effects of Azacytidine and Exenatide on cell viability and proliferation of human adipose tissue derived stem cells were analyzed with cytotoxicity assay. For differentiation procedure, of human adipose tissue derived stem cells were incubated with Azacytidine and Exenatide through four weeks. The morphological alterations of human adipose tissue derived stem cells were monitored and the expressions of cardiomyogenic differentiation markers (cTnI, GATA4 ve MYH7) were evaluated immunohistochemically. Also, cardiac troponin I (cTnI) levels in the cultures were measured using enzyme-linked immunosorbent assay. Results were evaluated by one way analysis of variance (ANOVA) and post-hoc test.

Results: Treatment of the human adipose tissue derived stem cells with Azacytidine significantly decreased cell viability (54.4%) compared to control whereas treatment of cells with Azacytidine + Exenatide prevented cell death in a dose-dependent manner. Cells treated with Azacytidine and Exenatide showed significant morphological alterations consistent with cardiyomyogenic differentiation, and increase in expression cardiomyogenic markers. cTnI levels were found significantly higher in cultures treated separately and together with Azacytidine and Exenatide compared to control.

Conclusion: Overall, these findings suggested that GLP-1 receptor agonist Exenatide may have beneficial effects on cardiomyogenic differention of human adipose tissue derived stem cells by reducing cell damage caused by Azacytidine.

Keywords

Glukagon benzeri peptit-1'in yağ doku kaynaklı mezenkimal kök hücrelerinin kardiyomiyositlere dönüşmesi üzerindeki etkisi

Öz

Amaç: Mezenkimal kök hücreler, çeşitli protokoller kullanılarak in vitro koşullarda kolaylıkla kardiyomiyositlere farklılaşabilir. Ancak bu protokollerde kullanılan ajanların hücre canlılığı üzerinde bazı olumsuz etkileri olduğu bildirilmiştir. Azasitidin mezenkimal kök hücreleri kalp kası hücrelerine farklandırmak için kullanılmaktadır. Bu çalışmanın amacı, bir GLP-1 reseptör agonisti olan Eksenatid'in insan yağ dokusu kaynaklı kök hücrelerinin kardiyomiyositlere farklılaşması ve canlılığı üzerindeki etkilerini araştırmaktır.

Gereç ve Yöntem: Azasitidin ve Eksenatid'in insan yağ doku kaynaklı mezenkimal kök hücreler üzerinde hücre canlılığı ve proliferasyonu üzerindeki etkileri ile sitotoksisite testleri yapıldı. Farklılanma protokolü için, hücreler dört hafta boyunca Azasitidin ve Eksenatid ile inkübe edildi. Hücrelerin morfolojik değişiklikleri izlendi ve kardiyomiyojenik farklılaşma belirteçlerinin (cTnI, GATA4 ve MYH7) ekspresyonları immünohistokimyasal olarak değerlendirildi. Ayrıca kültürlerdeki kardiyak troponin I (cTnI) seviyeleri enzime bağlı immünosorbent testi kullanılarak ölçüldü. Veriler, tek yönlü varyans analizi (ANOVA) ve post-hoc testi ile değerlendirildi.

Bulgular: İnsan yağ doku kaynaklı mezenkimal kök hücreler üzerine Azasitidin uygulaması, kontrole grubuna kıyasla hücre canlılığını önemli ölçüde azaltırken (%54.4) hücrelerin Azasitidin+Eksenatid ile uygulaması doza bağlı bir şekilde hücre ölümünü önledi. Azasitidin ve Eksenatid uygulanan hücreler, kardiyomiyojenik farklılaşma ile uyumlu önemli morfolojik değişiklikler ve kardiyomiyojenik belirteçlerde artış gösterdi. Ayrı ayrı ve birlikte uygulama yapılan gruplarda cTnI seviyeleri kontrole göre anlamlı derecede yüksek bulundu.

Sonuç: Bu bulgular GLP-1 reseptör agonisti Eksenatid'in, Azasitidin uygulamasının neden olduğu hücre hasarını azaltarak İnsan yağ doku kaynaklı mezenkimal kök hücrelerin kardiyomiyojenik farklılaşması üzerinde faydalı etkileri olabileceğini düşündürmektedir.

Anahtar Kelimeler

Kaynakça

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Ayrıntılar

Birincil Dil

İngilizce

Konular

Sağlık Kurumları Yönetimi

Bölüm

Araştırma Makalesi

Yazarlar

Özgür Tekin
0000-0003-3838-9990
Türkiye

Yiğit Uyanıkgil ^*
0000-0002-4016-0522
Türkiye

Dilek Taşkıran
0000-0002-4505-0939
Türkiye

Yayımlanma Tarihi

12 Aralık 2022

Gönderilme Tarihi

9 Mayıs 2022

Kabul Tarihi

7 Temmuz 2022

Yayımlandığı Sayı

Yıl 2022 Cilt: 61 Sayı: 4

DOI

https://doi.org/10.19161/etd.1180666

IZ

https://izlik.org/JA96PB48HB

Kaynak Göster

RIS / Bibtex

Vancouver

1.Özgür Tekin, Yiğit Uyanıkgil, Dilek Taşkıran. Effect of glucagon-like peptide-1 on differentiation of adipose derived mesenchymal stem cells into cardiomyocytes. ETD. 01 Aralık 2022;61(4):507-1. doi:10.19161/etd.1180666