Smad2, CYP19A, 卵巢颗粒细胞, 激活素 A, 卵泡液," /> Smad2, CYP19A, 卵巢颗粒细胞, 激活素 A, 卵泡液,"/> Smad2 ,CYP19A , Ovarian granulosa cells , Activin A, Follicular fluid,"/> <span style="line-height:2;font-size:14px;">激活素A 调节人卵巢颗粒细胞Smad2 和</span><span style="line-height:2;font-size:14px;">CYP19A 表达的研究</span>
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发育医学电子杂志  2019, Vol. 7 Issue (3): 196-202    DOI: 10.3969/j.issn.2095-5340.2019.03.007
  围产医学   论著 |产科 |
激活素A 调节人卵巢颗粒细胞Smad2 和CYP19A 表达的研究
刘超群 马会明 侯巧妮 何艳桃 杨玲玲 徐仙 王燕蓉 赵海燕 裴秀英
1. 宁夏医科大学;2. 北方民族大学
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
1. Ministry of Education/College of Basic Medicine Ningxia Medical University 2.North Minzu of University
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摘要 【摘要】 目的  探讨激活素A 通过Smad2 和CYP19A 表达促进颗粒细胞增殖的作用途径。 方法 2017 年3 ~ 8 月,在宁夏医科大学总医院生殖医学中心就诊的患者中,选取体外受精- 胚胎移植(IVFET)或卵胞质内单精子注射(ICSI)两种助孕方式治疗的15 例患者为研究对象。取分离纯化的卵巢颗粒细胞,经原代培养48 h 后分组:激活素A 组添加50 μg/ml 的激活素A,Bimagrumab 组添加5 μM 选择性ActR Ⅱ抑制剂Bimagrumab,混合组同时添加50 μg/ml 的激活素A 与20 μM 的Bimagrumab,再继续培养24 h。采用Western 印迹法检测蛋白表达水平,逆转录及实时PCR 检测mRNA 表达水平,细胞免疫荧光化学检测蛋白定位。采用单因素方差分析、t检验进行统计学分析。 结果  激活素A 组中ActR Ⅱ、p-Smad2/Smad2 蛋白表达水平均高于空白对照组,Bimagrumab 组中ActR Ⅱ和p-Smad2/Smad2 蛋白表达水平低于空白对照组,差异有统计学意义(P<0.05),提示Bimagrumab 对ActR Ⅱ有抑
制效果,激活素A 对ActR Ⅱ有促进效果。混合组中CYP19A1 蛋白表达低于空白对照组,激活素A组中CYP19A1 蛋白表达水平高于空白对照组;差异有统计学意义(P<0.05)。激活素A 组中Smad2 和CYP19A1 mRNA 表达水平高于空白对照组,混合组中Smad2 和CYP19A1 mRNA 表达水平低于空白对照组,差异均有统计学意义(P<0.05)。细胞免疫化学染色显示,Smad2 和CYP19A1 存在于卵巢颗粒细胞中,Smad2 阳性染色定位于细胞胞浆,CYP19A1 阳性染色定位于细胞胞浆和细胞膜。 结论 颗粒细胞分泌因子激活素 A 激活Act R Ⅱ信号通路,增加p-Smad2/Smad2 和CYP19A1 的表达水平,进一步促进颗粒细胞增殖和分化。
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关键词:  Smad2')" href="#">Smad2  CYP19A  卵巢颗粒细胞  激活素 A  卵泡液    
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
Key words:  Smad2 ')" href="#">Smad2    CYP19A    Ovarian granulosa cells    Activin A    Follicular fluid
收稿日期:  2018-07-19                出版日期:  2019-07-30      发布日期:  2019-07-26      期的出版日期:  2019-07-30
基金资助: 国家自然科学基金(81560261,81660813)
通讯作者:  裴秀英https://baike.baidu.com/item/%E8%A3%B4%E7%A7%80%E8%8B%B1/20611827?fr=aladdin 赵海燕    E-mail:  peixiuying@163.com, hy_zhao06@163.com
引用本文:    
刘超群 马会明 侯巧妮 何艳桃 杨玲玲 徐仙 王燕蓉 赵海燕 . 激活素A 调节人卵巢颗粒细胞Smad2 和CYP19A 表达的研究[J]. 发育医学电子杂志, 2019, 7(3): 196-202.
LIU Chao-qun, MA Hui-ming, HOU Qiao-ni, HE Yan-tao, YANG Ling-ling, XU xian, WANG Yanrong. Research of activin A on modulation of the expression of Smad2 and CYP19A in human ovarian granulosa cells. Journal of Developmental Medicine(Electronic Version), 2019, 7(3): 196-202.
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http://www.fyyxzz.com/CN/10.3969/j.issn.2095-5340.2019.03.007  或          http://www.fyyxzz.com/CN/Y2019/V7/I3/196
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