發(fā)布時(shí)間：2018-04-05 16:28

  本文選題：多囊卵巢綜合征　切入點(diǎn)：顆粒細(xì)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(jiàn)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(shí)驗(yàn)通過(guò)研究糖原合成酶激酶-3(Glycogen synthase kinese-3,GSK-3)抑制劑SB216763對(duì)PCOS卵巢顆粒細(xì)胞AKT/GSK-3通路的影響,探討PCOS卵巢局部IR發(fā)生的可能機(jī)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(shí)伴有PCOS患者的卵泡黃素化顆粒細(xì)胞(Granulosa cell,GC),行體外細(xì)胞培養(yǎng)(n=20),把每份顆粒細(xì)胞樣本分別接種在兩個(gè)培養(yǎng)瓶中,其中一個(gè)培養(yǎng)瓶中的細(xì)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(duì)照組,記為N-PCOS組(n=16)。采用蛋白免疫印跡試驗(yàn)(Western Blot試驗(yàn))檢測(cè)各實(shí)驗(yàn)組顆粒細(xì)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(dá)情況。結(jié)果:t檢驗(yàn)統(tǒng)計(jì)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(dòng)量、Gn總用量均無(wú)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(xì)胞中GSK-3、p-GSK-3β(Ser9)表達(dá)量較N-PCOS組低,而p-IRS-1(Ser312)表達(dá)量較N-PCOS組高,差異均有統(tǒng)計(jì)學(xué)意義(P0.05);經(jīng)SB216763處理后的PCOS+IN組p-GSK-3β(Ser9)表達(dá)量仍較N-PCOS組低,但較PCOS組高,p-IRS-1(Ser312)表達(dá)量較N-PCOS組高,但較PCOS組低,差異均具有統(tǒng)計(jì)學(xué)意義(P0.05);SB216763處理后GSK-3表達(dá)量與PCOS組比較無(wú)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(xì)胞中p-GSK-3β表達(dá)量降低,表現(xiàn)為AKT/GSK-3途徑活性的增強(qiáng);卵巢顆粒細(xì)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(xì)胞中GSK-3的活性減弱,降低p-IRS-1的表達(dá),在一定程度上抑制MAPK通路的活化對(duì)糖代謝途徑的負(fù)反饋?zhàn)饔?改善卵巢局部IR,為臨床治療提供科學(xué)的靶向點(diǎn)。
[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(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R711.75

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