Cx43在小鼠卵巢中的表达及黄体生成素和双氢睾酮对小鼠卵巢和大鼠颗粒细胞Cx43表达的影响

发布时间：2018-03-07 13:33

  本文选题：黄体生成素　切入点：双氢睾酮　出处：《宁夏医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：缝隙连接蛋白43(Connexin43,Cx43)是哺乳动物卵巢卵泡颗粒细胞间表达最多的一种缝隙连接蛋白,在卵泡发育、卵母细胞生长等环节中发挥重要作用。卵泡颗粒细胞Cx43的表达受促性腺激素的调节,已经发现卵泡刺激素(Follicle stimulating hormone,FSH)可促进发育卵泡颗粒细胞Cx43的表达,加强颗粒细胞之间的缝隙连接。黄体生成素(Luteinizing hormone,LH)是否具有促进Cx43表达的作用以及具体的作用机理,尚未明确。此外,雄激素在卵泡发育和女性生育调节过程中同样发挥重要作用,而雄激素对颗粒细胞Cx43表达的影响亦不明确。本研究以不同发育阶段的雌性C57小鼠和卵巢以及原代培养的大鼠颗粒细胞为材料,应用形态学,细胞生物学和分子生物学方法,研究小鼠生长发育过程中卵巢Cx43表达的模式,以及LH和双氢睾酮(Dihydrotestosterone,DHT)对小鼠卵巢和大鼠颗粒细胞Cx43表达的影响及可能的机理。实验结果显示,在小鼠卵巢中,Cx43表达于卵巢颗粒细胞胞膜上,LHR表达在卵泡膜细胞、颗粒细胞及卵母细胞胞质中。随卵泡发育,颗粒细胞数量的增加,Cx43及LHR的表达明显增强。使用不同浓度的LH及DHT体外孵育小鼠卵巢,发现LH及DHT均可促进小鼠卵巢Cx43的表达。对于原代培养21d大鼠颗粒细胞,Cx43表达于颗粒细胞胞质和部分细胞膜上。LH和Wnt/β-catenin信号通路激动剂LiCl均可增加颗粒细胞Cx43的表达,而使用Wnt/β-catenin信号通路抑制剂FH535,可减少LH诱导的Cx43表达。同样,使用DHT和Wnt/β-catenin信号通路激动剂LiCl均可增加颗粒细胞Cx43的表达,使用Wnt/β-catenin信号通路抑制剂FH535后,可减少DHT诱导的Cx43表达。综上所述,我们得到如下结论:(1)小鼠自然生长状态下,随卵泡发育,颗粒细胞数量增加,Cx43及LHR表达明显增强;LH可能通过LHR调控卵泡颗粒细胞Cx43的表达。(2)适合浓度LH和DHT体外干预小鼠卵巢组织,可促进卵泡颗粒细胞Cx43表达,说明LH和DHT是Cx43表达的调控因素。(3)LH和DHT均可通过激活Wnt/β-catenin信号通路促进颗粒细胞Cx43表达。
[Abstract]:Gap junction protein 43 (Connexin 43) is the most expressed gap junction protein in follicular granulosa cells of mammalian ovary. The expression of Cx43 in follicular granulosa cells is regulated by gonadotropin. It has been found that Follicle stimulating stimulating FSHs can promote the expression of Cx43 in follicular granulosa cells. It is not clear whether luteinizing hormone luteinizing hormone LHH can promote the expression of Cx43 and its specific mechanism. Androgen also plays an important role in follicular development and the regulation of women's fertility. The effect of androgen on the expression of Cx43 in granulosa cells was also unclear. In this study, morphological, cellular and molecular biological methods were used to study the effects of androgen on the expression of Cx43 in female C57 mice and ovary and primary cultured rat granulosa cells in different developmental stages. To study the expression pattern of Cx43 in mouse ovary and the effect of LH and dihydrotestosterone on the expression of Cx43 in mouse ovary and rat granulosa cells. In mouse ovary, Cx43 was expressed on the membrane of granulosa cells, and LHR was expressed in the cytoplasm of follicular membrane cells, granulosa cells and oocytes. The expression of Cx43 and LHR was significantly increased with the increase of the number of granulosa cells. The ovaries of mice were incubated with different concentrations of LH and DHT in vitro. It was found that both LH and DHT could promote the expression of Cx43 in mouse ovary, and that the expression of Cx43 in granulosa cells was increased in primary cultured rat granulosa cells on day 21, and the expression of Cx43 in granulosa cells was increased by LiCl, a signal pathway agonist of Wnt/ 尾 -catenin and LH on the cytoplasm of granulosa cells. Using Wnt/ 尾 -catenin signal pathway inhibitor FH 535 can reduce LH induced Cx43 expression. Similarly, DHT and Wnt/ 尾 -catenin signal pathway agonist LiCl can increase the expression of Cx43 in granulosa cells, and Wnt/ 尾 -catenin signal pathway inhibitor FH535 can increase the expression of Cx43 in granulosa cells. The expression of Cx43 induced by DHT can be reduced. To sum up, we can draw the following conclusion: 1) under the natural growth state of mice, the follicle develops with the development of follicle. The increased expression of Cx43 and LHR in granulosa cells may regulate the expression of Cx43 in granulosa cells by LHR. It may be suitable for LH and DHT to interfere with mouse ovarian tissue in vitro and promote the expression of Cx43 in granulosa cells of follicle. These results suggest that LH and DHT are regulatory factors of Cx43 expression. LH and DHT can promote Cx43 expression in granulosa cells by activating Wnt/ 尾 -catenin signaling pathway.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R339.22

【参考文献】

相关期刊论文 前4条

1 张焕焕;魏兆莲;;脱氢表雄酮治疗卵巢储备功能低下的不孕患者研究进展[J];中华全科医学;2016年01期

2 范梦夏;乔洁;;黄体生成素受体mRNA结合蛋白的研究进展[J];上海交通大学学报(医学版);2015年06期

3 安锦霞;倪亚莉;;雄激素在卵巢储备功能低下妇女中的研究进展[J];国际生殖健康/计划生育杂志;2013年01期

4 谭萍;王满意;袁小丽;白汝岚;张家骅;;连接蛋白Cx43和Cx37在卵泡发育中的作用[J];动物医学进展;2007年05期

相关博士学位论文 前1条

1 闫真;间隙连接蛋白在绵羊早期胚胎中的表达和作用研究[D];内蒙古大学;2009年

相关硕士学位论文 前4条

1 闫胜男;缝隙连接蛋白Cx43在人牙髓组织中表达的免疫组化研究[D];吉林大学;2016年

2 文欢;结直肠癌细胞与成纤维细胞相互作用过程中成纤维细胞生物学行为和Cx43表达研究[D];浙江大学;2015年

3 李夏薇;醛固酮对H9C2心肌细胞缝隙连接蛋白43的影响[D];南昌大学;2012年

4 张晓光;促性腺激素对小鼠卵巢VEGF、Survivin的表达及玻璃化冻存移植后产子率的影响[D];宁夏医科大学;2012年

，

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