Akif Hakan KURT, Rukiye Nalan TİFTİK, İsmail ÜN, Kansu BÜYÜKAFŞAR

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

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.

Anahtar Kelimeler:

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

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

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,

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