GSK-3抑制剂SB216763对多囊卵巢综合征颗粒细胞GSK-3、IRS-1表达的影响

发布时间：2018-04-05 16:28

本文选题：多囊卵巢综合征　切入点：颗粒细胞　出处：《青岛大学》2017年硕士论文

【摘要】：目的:多囊卵巢综合征(Polycystic ovarian syndrome,PCOS)是一种较常见的女性生殖功能障碍性疾病,其发病相关因素以胰岛素抵抗(IR)为主。本实验通过研究糖原合成酶激酶-3(Glycogen synthase kinese-3,GSK-3)抑制剂SB216763对PCOS卵巢颗粒细胞AKT/GSK-3通路的影响,探讨PCOS卵巢局部IR发生的可能机制。方法:收集本院因不孕而要求行体外受精/卵胞浆内单精子注射-胚胎移植(IVF/ICSI-ET)助孕,同时伴有PCOS患者的卵泡黄素化颗粒细胞(Granulosa cell,GC),行体外细胞培养(n=20),把每份颗粒细胞样本分别接种在两个培养瓶中,其中一个培养瓶中的细胞用GSK-3抑制剂SB216763处理,记为PCOS+IN组,未加抑制剂者记为PCOS组。另外收集因不孕而要求行IVF/ICSI-ET助孕,非PCOS患者的卵泡黄素化GC作为对照组,记为N-PCOS组(n=16)。采用蛋白免疫印迹试验(Western Blot试验)检测各实验组颗粒细胞中GSK-3、磷酸化GSK-3β(p-GSK-3β(Ser9))以及磷酸化胰岛素受体底物-1(p-IRS-1(Ser312))表达情况。结果:t检验统计分析PCOS组与非PCOS组基本特征提示两组患者平均年龄(28±6)、不孕年限、体重指数(BMI)、基础雌二醇(bE2)、卵泡刺激素(bFSH)、泌乳素(bPRL)、Gn启动量、Gn总用量均无明显差异(P0.05)。PCOS组的基础促黄体生成激素(bLH)、睾酮(bT)、获卵数明显高于N-PCOS组(P0.05)。单因素方差分析示PCOS组患者颗粒细胞中GSK-3、p-GSK-3β(Ser9)表达量较N-PCOS组低,而p-IRS-1(Ser312)表达量较N-PCOS组高,差异均有统计学意义(P0.05);经SB216763处理后的PCOS+IN组p-GSK-3β(Ser9)表达量仍较N-PCOS组低,但较PCOS组高,p-IRS-1(Ser312)表达量较N-PCOS组高,但较PCOS组低,差异均具有统计学意义(P0.05);SB216763处理后GSK-3表达量与PCOS组比较无明显差异(P0.05)。结论:PCOS患者与非PCOS患者相比,其卵巢颗粒细胞中p-GSK-3β表达量降低,表现为AKT/GSK-3途径活性的增强;卵巢颗粒细胞中IRS-1/MAPK途径与AKT/GSK-3途径存在一定的相互联系,GSK-3抑制剂SB216763使用后可使颗粒细胞中GSK-3的活性减弱,降低p-IRS-1的表达,在一定程度上抑制MAPK通路的活化对糖代谢途径的负反馈作用,改善卵巢局部IR,为临床治疗提供科学的靶向点。
[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 卤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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.75

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