Sıçan Koroner Mikrovasküler Endotel Hücre Kültüründe Östrojenin Oluşturduğu RhoA Up-regülasyonu Üzerine ICI182,780, Progesteron ve Testosteronun Etkisi

Year 2011, Volume: 4 Issue: 2, 17 - 21, 01.06.2011

Akif Hakan Kurt Rukiye Nalan Tiftik İsmail Ün Kansu Büyükafşar

https://izlik.org/JA37YD69PR

Abstract

Abstract The Effects of ICI182,780, Progesterone and Testosterone on Estrogen Induced Upregulation of RhoA Expressions in The Rat Coronary Microvascular Endothelium Cell Culture Method: Wistar rats were used in the study. Coronary microvascular endothelial cells were obtained by the mounted rat hearts on a constant-flow Langendorff system. The cell suspensions were plated and incubated at 37oC under 5% CO2. Confluent flasks were passaged. The third passage coronary microvascular endothelial cells were made quiscent 24 hours before experimentation. Next, it was incubated with either 17β-estradiol (10-8 -10-7M), or its combination with ICI182, 780 (10-5M), progesterone (10-8 - 10-7M), and testosterone (10-8 - 10-7M) for 24 hrs. Afterwards, the cells in flasks were homogenized and RhoA protein enzyme expression levels were measured by Western-Blotting technique. Results: 17-β estradiol causes RhoA up-regulation that can not be blocked by progesterone and testosterone in rat coronary microvascular endothelial cell culture. Conclusion: Activation of Rho/Rho-kinase pathway might be responsible for the development of estrogen associated cardiovascular complications.

Keywords

endothelium; 17β-estradiol; progesterone; rhoA; testosterone

Sıçan Koroner Mikrovasküler Endotel Hücre Kültüründe Östrojenin Oluşturduğu RhoA Up-regülasyonu Üzerine ICI182,780, Progesteron ve Testosteronun Etkisi

https://izlik.org/JA37YD69PR

Abstract

Türkçe Özet: Amaç: Bu çalışmada sıçan koroner mikrovasküler endotel hücrelerinde tek başına estradiol ve estradiolün progesteron, testosteron ve estrojen reseptör α ve β blokörü ile kombinasyonlarının RhoA ekspresyonu üzerine etkileri Western blot analizi ile değerlendirildi. Yöntem: Çalışmada Wistar türü sıçanlar kullanıldı. Kalpler Langendorff kalp perfüzyon sistemine asılarak koroner mikrovasküler endotel hücreleri elde edildi. Hücreler kültür flasklarına ekilerek 37ºC\'de %5 CO2\'li ortamda inkübe edildi. Konfluent olan flasklar pasajlanarak çoğaltıldı. 3. pasaj koroner mikrovasküler endotel hücreleri, deneylerden 24 saat önce bölünmeleri durdurulup 17β-estradiol (10-8 ve 10-7M) tek başına veya 17β-estradiol testosteron (10-8 ve 10-7M), progesteron (10-8 ve 10-7M) ve ICI 182, 780 (10-5M), kombinasyonu ilave edilerek 24 saat inkübe edildi. İnkübasyon sonrası flasklardaki hücreler homojenize edilerek Western-Blot yöntemi ile RhoA protein enzim ekspresyon düzeyleri ölçüldü. Bulgular: 17β-estradiol sıçan koroner mikrovasküler endotel hücre kültüründe progesteron ve testosteron ile bloke edilemeyen RhoA up-regülasyonu neden olmaktadır. Sonuç: Sonuç olarak östrojene bağlı olarak gözlemlenen kardiyovasküler komplikasyonlardan Rho/Rho-kinaz yolağının aktivasyonu sorumlu olabilir.

Keywords

endotel; 17β-estradiol; progesteron; rhoA; testosteron

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