Journal of Developmental Medicine(Electronic Version)  2019, Vol. 7 Issue (3): 196-202 DOI: 10.3969/j.issn.2095-5340.2019.03.007 |
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| Research of activin A on modulation of the expression of Smad2 and CYP19A in human ovarian granulosa cells |
| LIU Chao-qun MA Hui-ming, HOU Qiao-ni, HE Yan-tao, YANG Ling-ling, XU xian, WANG Yanrong,ZHAO Hai-yan, PEI Xiu-ying
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| 1. Ministry of Education/College of Basic Medicine Ningxia Medical University 2.North Minzu of University |
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Abstract 【Abstract】 Objective To explore the effect of activin A to promote the proliferation of granulosa cells through the expression of Smad2 and CYP19A. Methods From March to August 2017, 15 cases were included who were treated by in vitro fertilization and embryo transfer(IVF-ET) and intracytoplasmicsperm injection (ICSI) in the center of reproductive medicine of General Hospital of Ningxia Medical University. The granulosa cells with isolating from follicular fluid were selected as the research object.After incubation for 48 hours, 50 μg / ml activin A was added to activin A group, 5 μM selective ActR
Bimagrumab Ⅱ inhibitors was added to Bimagrumab group, 50 μg / ml activin A and 20 μM Bimagrumab were added to hybrid group, all of those were incubated for 24 h. The protein expression level was detected by Western blot assay. The mRNA expression level was detected by reverse transcription and real-time PCR. The protein localization was detected by immune-fluorescence cytochemistry technology. One-Way ANOVA and t test were used for statistical analysis. Results The protein expression level of ActR Ⅱ , p-Smad2/Smad2 in activin A group was higher than that in blank control group, and the protein expression level of ActR Ⅱ , p-Smad2/Smad2 in Bimagrumab group was lower than that in blank control group, which was statistical significance (P<0.05). It showed Bimagrumab has inhibited effect for ActR Ⅱ and activin A has promotional effect for ActR Ⅱ . The protein expression of CYP19A1 in hybrid group was lower than that in blank control group , the protein expression level of CYP19A1 in activin A group was higher than that in blank control group, which was statistical significance (P<0.05).The expression level of Smad2 and CYP19A1 mRNA in activin A group was higher than that in blank control group. The expression
level of Smad2 and CYP19A1 mRNA in hybrid group was lower than that in blank control group. There were all statistical significance (P<0.05). Immunocytochemical staining technique showed Smad2 and CYP19A1 existed in granulosa cells of ovary, positive staining of Smad 2 was located in cytoplasm, positive staining of CYP19A1 was located in cytoplasm and cell membranes. Conclusion Activin A of granulosa cells can activate signaling pathways of ActR Ⅱ and improve the expression level of p-Smad2/Smad2 and CYP19A1, which can further enhance the proliferation, and differentiation of granulosa cells
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Received: 19 July 2018
Published: 26 July 2019
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