Preadipositlerde (3T3-L1) Östrojenin Adiposit Diferansiyasyonu Üzerine Etkisi: Rho/Rho-kinaz Yolağının Olası Katkısı

Abstract

Abstract Effects of Estrogen on Adipocyte Diferentiation in Preadipocyte Cell Line (3T3-L1): Possible Involvement of Rho/Rho-Kinase Signaling Aim: In this study, it was aimed to investigate potential effects of estrogen on the differantiation of preadipocytes (3T3-L1) to mature adipocytes and the possible involvement of Rho/Rho-kinase pathway in this matter. Method: 3T3-L1 cell suspensions were plated and incubated at 37oC under 5% CO2. Confluent flasks were passaged. Next, 3T3-L1 cells were treated with estradiol at various concentrations (10-9, 10-8, 10-7, 10-6, 10-5 and 10-4M) for a period of 0-2 and 0-8 days. The experiment was concluded on the day of 8. Adipocyte differentiation was assessed by lipid-specific Oil Red O staining. The Oil Red O nontreated cells were homogenized. Subsequently, RhoA, ROCK-1 and ROCK-2 protein expression levels in these cells were measured by Western-Blotting technique. Results: When applied in the beginning of the differentiation process (the 2nd day), 17β-estradiol decrease the differentiation. On the contrary, it becomes ineffective when applied for 8 days. In addition, 17β-estradiol did not significantly change ROCK-1, ROCK-2 and RhoA protein expression levels. Conclusion: 17β-estradiol, the main estrogenic hormone, has diverse effects on adipocyte differentiation at different stages of differentiation. However, this impact does not appear to be mediated via Rho/Rho-kinase pathway.

Keywords

• adipocyte differentiation; estrogen; rho-kinase

Preadipositlerde (3T3-L1) Östrojenin Adiposit Diferansiyasyonu Üzerine Etkisi: Rho/Rho-kinaz Yolağının Olası Katkısı

Abstract

Türkçe Özet: Amaç: Bu çalışmada estrojenin, preadipositlerin (3T3-L1) olgun yağ hücrelerine diferansiyasyonuna olan etkilerinin incelenmesi ve ayrıca bu etkilere, eğer varsa, Rho/Rho kinaz yolağının katkısını araştırılması amaçlandı. Yöntem: 3T3-L1 hücreleri kültür flasklarına ekilerek 37ºC\'de %5 CO2\'li ortamda inkübe edildi. Konfluent olan flasklar pasajlanarak çoğaltıldı. Estradiol 10-9, 10-8, 10-7, 10-6, 10-5 ve 10-4M konsantrasyonda 3T3-L1 hücrelerine 2 ve 8 gün boyunca uygulandı. 8. günde deney sonlandırıldı ve adiposit diferansiyasyonu Oil Red O boyama ile değerlendirildi. Oil Red uygulanmayan hücreler ise homojenize edilerek Western-Blot yöntemi ile RhoA, ROCK-1 ve ROCK-2 protein ekspresyon düzeyleri ölçüldü. Bulgular: 17β-estradiol diferansiyasyon sürecinin başlangıcında (2. günde) uygulandığında diferansiyasyonu azaltmaktadır. Buna karşılık 8 gün boyunca uygulandığında ise etkisiz bulunmuştur. Ayrıca 17β-estradiol RhoA, ROCK-1 ve ROCK-2 protein ekspresyon düzeylerini anlamlı olarak değiştirmedi. Sonuç: 17β-estradiol, preadiposit diferansiyasyon sürecine farklı zaman dilimlerinde farklı etkilerle katkı sağlamaktadır. Ancak, bu etki Rho/Rho-kinaz yolağı aracılı görünmemektedir.

Keywords

• adiposit diferansiyasyonu; östrojen; rho-kinaz

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