本文選題：黃體生成素　切入點(diǎn)：雙氫睪酮　出處：《寧夏醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：縫隙連接蛋白43(Connexin43,Cx43)是哺乳動(dòng)物卵巢卵泡顆粒細(xì)胞間表達(dá)最多的一種縫隙連接蛋白,在卵泡發(fā)育、卵母細(xì)胞生長(zhǎng)等環(huán)節(jié)中發(fā)揮重要作用。卵泡顆粒細(xì)胞Cx43的表達(dá)受促性腺激素的調(diào)節(jié),已經(jīng)發(fā)現(xiàn)卵泡刺激素(Follicle stimulating hormone,FSH)可促進(jìn)發(fā)育卵泡顆粒細(xì)胞Cx43的表達(dá),加強(qiáng)顆粒細(xì)胞之間的縫隙連接。黃體生成素(Luteinizing hormone,LH)是否具有促進(jìn)Cx43表達(dá)的作用以及具體的作用機(jī)理,尚未明確。此外,雄激素在卵泡發(fā)育和女性生育調(diào)節(jié)過程中同樣發(fā)揮重要作用,而雄激素對(duì)顆粒細(xì)胞Cx43表達(dá)的影響亦不明確。本研究以不同發(fā)育階段的雌性C57小鼠和卵巢以及原代培養(yǎng)的大鼠顆粒細(xì)胞為材料,應(yīng)用形態(tài)學(xué),細(xì)胞生物學(xué)和分子生物學(xué)方法,研究小鼠生長(zhǎng)發(fā)育過程中卵巢Cx43表達(dá)的模式,以及LH和雙氫睪酮(Dihydrotestosterone,DHT)對(duì)小鼠卵巢和大鼠顆粒細(xì)胞Cx43表達(dá)的影響及可能的機(jī)理。實(shí)驗(yàn)結(jié)果顯示,在小鼠卵巢中,Cx43表達(dá)于卵巢顆粒細(xì)胞胞膜上,LHR表達(dá)在卵泡膜細(xì)胞、顆粒細(xì)胞及卵母細(xì)胞胞質(zhì)中。隨卵泡發(fā)育,顆粒細(xì)胞數(shù)量的增加,Cx43及LHR的表達(dá)明顯增強(qiáng)。使用不同濃度的LH及DHT體外孵育小鼠卵巢,發(fā)現(xiàn)LH及DHT均可促進(jìn)小鼠卵巢Cx43的表達(dá)。對(duì)于原代培養(yǎng)21d大鼠顆粒細(xì)胞,Cx43表達(dá)于顆粒細(xì)胞胞質(zhì)和部分細(xì)胞膜上。LH和Wnt/β-catenin信號(hào)通路激動(dòng)劑LiCl均可增加顆粒細(xì)胞Cx43的表達(dá),而使用Wnt/β-catenin信號(hào)通路抑制劑FH535,可減少LH誘導(dǎo)的Cx43表達(dá)。同樣,使用DHT和Wnt/β-catenin信號(hào)通路激動(dòng)劑LiCl均可增加顆粒細(xì)胞Cx43的表達(dá),使用Wnt/β-catenin信號(hào)通路抑制劑FH535后,可減少DHT誘導(dǎo)的Cx43表達(dá)。綜上所述,我們得到如下結(jié)論:(1)小鼠自然生長(zhǎng)狀態(tài)下,隨卵泡發(fā)育,顆粒細(xì)胞數(shù)量增加,Cx43及LHR表達(dá)明顯增強(qiáng);LH可能通過LHR調(diào)控卵泡顆粒細(xì)胞Cx43的表達(dá)。(2)適合濃度LH和DHT體外干預(yù)小鼠卵巢組織,可促進(jìn)卵泡顆粒細(xì)胞Cx43表達(dá),說明LH和DHT是Cx43表達(dá)的調(diào)控因素。(3)LH和DHT均可通過激活Wnt/β-catenin信號(hào)通路促進(jìn)顆粒細(xì)胞Cx43表達(dá)。
[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.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R339.22

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