本文選題：多囊卵巢綜合征　切入點：顆粒細胞　出處：《青島大學(xué)》2017年碩士論文

【摘要】：目的:多囊卵巢綜合征(Polycystic ovarian syndrome,PCOS)是一種較常見的女性生殖功能障礙性疾病,其發(fā)病相關(guān)因素以胰島素抵抗(IR)為主。本實驗通過研究糖原合成酶激酶-3(Glycogen synthase kinese-3,GSK-3)抑制劑SB216763對PCOS卵巢顆粒細胞AKT/GSK-3通路的影響,探討PCOS卵巢局部IR發(fā)生的可能機制。方法:收集本院因不孕而要求行體外受精/卵胞漿內(nèi)單精子注射-胚胎移植(IVF/ICSI-ET)助孕,同時伴有PCOS患者的卵泡黃素化顆粒細胞(Granulosa cell,GC),行體外細胞培養(yǎng)(n=20),把每份顆粒細胞樣本分別接種在兩個培養(yǎng)瓶中,其中一個培養(yǎng)瓶中的細胞用GSK-3抑制劑SB216763處理,記為PCOS+IN組,未加抑制劑者記為PCOS組。另外收集因不孕而要求行IVF/ICSI-ET助孕,非PCOS患者的卵泡黃素化GC作為對照組,記為N-PCOS組(n=16)。采用蛋白免疫印跡試驗(Western Blot試驗)檢測各實驗組顆粒細胞中GSK-3、磷酸化GSK-3β(p-GSK-3β(Ser9))以及磷酸化胰島素受體底物-1(p-IRS-1(Ser312))表達情況。結(jié)果:t檢驗統(tǒng)計分析PCOS組與非PCOS組基本特征提示兩組患者平均年齡(28±6)、不孕年限、體重指數(shù)(BMI)、基礎(chǔ)雌二醇(bE2)、卵泡刺激素(bFSH)、泌乳素(bPRL)、Gn啟動量、Gn總用量均無明顯差異(P0.05)。PCOS組的基礎(chǔ)促黃體生成激素(bLH)、睪酮(bT)、獲卵數(shù)明顯高于N-PCOS組(P0.05)。單因素方差分析示PCOS組患者顆粒細胞中GSK-3、p-GSK-3β(Ser9)表達量較N-PCOS組低,而p-IRS-1(Ser312)表達量較N-PCOS組高,差異均有統(tǒng)計學(xué)意義(P0.05);經(jīng)SB216763處理后的PCOS+IN組p-GSK-3β(Ser9)表達量仍較N-PCOS組低,但較PCOS組高,p-IRS-1(Ser312)表達量較N-PCOS組高,但較PCOS組低,差異均具有統(tǒng)計學(xué)意義(P0.05);SB216763處理后GSK-3表達量與PCOS組比較無明顯差異(P0.05)。結(jié)論:PCOS患者與非PCOS患者相比,其卵巢顆粒細胞中p-GSK-3β表達量降低,表現(xiàn)為AKT/GSK-3途徑活性的增強;卵巢顆粒細胞中IRS-1/MAPK途徑與AKT/GSK-3途徑存在一定的相互聯(lián)系,GSK-3抑制劑SB216763使用后可使顆粒細胞中GSK-3的活性減弱,降低p-IRS-1的表達,在一定程度上抑制MAPK通路的活化對糖代謝途徑的負反饋作用,改善卵巢局部IR,為臨床治療提供科學(xué)的靶向點。
[Abstract]:Objective: polycystic ovarian syndrome (PCOS) is a common disease of female reproductive dysfunction.In this study, we investigated the effect of glycogen synthase kinase (Glycogen synthase kinese-3) inhibitor SB216763 on the AKT/GSK-3 pathway of PCOS ovarian granulosa cells, and explored the possible mechanism of local IR in PCOS ovary.Methods: IVF / ICSI-ETT (IVF / ICSI-ETT) was performed in our hospital for infertility.Follicular luteinized granulosa granulosa cells with PCOS were cultured in vitro. Each sample of granulosa cells was inoculated into two culture flasks, one of which was treated with GSK-3 inhibitor SB216763 and recorded as PCOS IN group.Those without inhibitor were recorded as PCOS group.In addition, the follicular luteinized GC of non-#en1# patients was taken as the control group and recorded as N-PCOS group.The expression of GSK-3, phosphorylated GSK-3 尾 -p-GSK-3 尾 -Ser9N and phosphorylated insulin receptor substrate--1p-IRS-1 Ser312A) in granulosa cells of each experimental group were detected by Western blot assay (Western Blot).Results the statistical analysis of PCOS group and non-#en1# group showed that the average age of the two groups was 28 鹵6, and the infertile years.Univariate ANOVA analysis showed that the expression of GSK-3P- GSK-3 尾 -ser9) in granulosa cells of PCOS group was lower than that in N-PCOS group, while the expression of p-IRS-1 + Ser312) was higher than that in N-PCOS group (P 0.05), and the expression of p-GSK-3 尾 -Ser9 in PCOS IN treated with SB216763 was still lower than that in N-PCOS group.However, the expression of p-IRS-1 ser312 in PCOS group was higher than that in N-PCOS group, but lower than that in PCOS group. There was no significant difference in the expression of GSK-3 between PCOS group and PCOS group.Conclusion the expression of p-GSK-3 尾 in granulosa cells of ovarian granulosa cells in patients with PCOS is lower than that in non-PCOS patients, and the expression of p-GSK-3 尾 in ovarian granulosa cells is increased by increasing the activity of AKT/GSK-3 pathway.There is a certain correlation between IRS-1/MAPK pathway and AKT/GSK-3 pathway in granulosa cells. SB216763, a inhibitor of GSK-3, can weaken the activity of GSK-3 and decrease the expression of p-IRS-1 in granulosa cells.To some extent inhibit the activation of MAPK pathway on the negative feedback of glucose metabolism pathway improve the ovarian local IRR and provide a scientific target for clinical treatment.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R711.75

【參考文獻】

相關(guān)期刊論文 前10條

1 任文超;劉建新;祁秀娟;丁鈺;王琳;;二甲雙胍對卵巢顆粒細胞IRS-1及ERK-2基因表達的影響[J];國際生殖健康/計劃生育雜志;2013年03期

2 時照明;吳道愛;張士榮;周靜;孫衛(wèi)華;席玉玲;張曉梅;;多囊卵巢綜合征患者肥胖、高胰島素和高雄激素血癥的相關(guān)性研究[J];中國糖尿病雜志;2012年07期

3 林瑩瑩;孫寶治;李曉芳;;多囊卵巢綜合征病人血清CRP、內(nèi)脂素和chemerin變化及其意義[J];齊魯醫(yī)學(xué)雜志;2012年03期

4 張慧英;張艷芳;韓玉];薛鳳霞;趙曉徽;張秀蘭;;多囊卵巢綜合征患者子宮內(nèi)膜組織中胰島素PI3K/Akt信號通路的活化及其意義[J];中華婦產(chǎn)科雜志;2012年01期

5 梁晨;叢晶;�；�;匡洪影;侯麗輝;吳效科;;補腎化痰復(fù)方對多囊卵巢綜合征模型大鼠卵巢內(nèi)胰島素信號傳導(dǎo)分子的調(diào)控[J];中國中西醫(yī)結(jié)合雜志;2011年12期

6 張艷芳;韓玉];;多囊卵巢綜合征胰島素抵抗與信號通路缺陷[J];國際生殖健康/計劃生育雜志;2010年04期

7 基亞雷利;李力華;;兒童肥胖與胰島素抵抗[J];糖尿病天地(臨床);2009年05期

8 范樂平;朱杰;;多囊卵巢綜合征的胰島素抵抗機制研究進展[J];醫(yī)學(xué)綜述;2009年09期

9 韓曉芬;鐘雪梅;;多囊卵巢綜合征胰島素抵抗的研究進展[J];西南軍醫(yī);2009年01期

10 胡敏;李威;吳效科;;肥胖對多囊卵巢綜合征患者生殖內(nèi)分泌功能的影響[J];醫(yī)學(xué)研究生學(xué)報;2008年11期

，

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