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Contractility Measurements of Human Uterine Smooth Muscle to Aid Drug Development
Published on: January 26, 2018
Endothelin-1-induced expression of α-smooth muscle actin in human myometrial fibroblasts
Zhaoning Duan1, Ying Jia1, Jing Zhang1
1Department of Gynaecology and Obstetrics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Aim:
We aimed to investigate the potential role of endothelin-1 (ET-1) in the regulation of the expression of α-smooth muscle actin (α-SMA) in human myometrial fibroblasts.
Methods:
Primary myometrial fibroblasts were obtained from myometrium and were identified by immunocytochemical staining. Then, 1 × 107 cells were treated with ET-1 at a concentration of 0.1, 1.0, 10.0, or 100.0 nM for 24 h. To investigate the time course effects of ET-1 on the growth of fibroblasts, 1 × 107 cells were treated with 10.0 nM ET-1 for 6, 12, 24, and 48 h. Real-time quantitative PCR (qPCR) and Western blot analysis were used to determine the expression of α-SMA mRNA and protein, respectively.
Results:
Human myometrial fibroblasts were identified by immunohistochemistry. Compared with the control group, the expression levels of α-SMA mRNA and protein were identified in cells treated with ET-1 at a concentration of 0.1, 1.0, 10.0, or 100.0 nM for 24 h (P < 0.05). ET-1 treatment affected the expression of α-SMA mRNA and protein in a dose-dependent manner (P < 0.05). The induction of α-SMA mRNA and protein expression increased from 6 to 48 h.
Conclusion:
The results show that ET-1 induces the expression of α-SMA in human myometrium in vitro.
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