摘要
目的研究肥胖与非肥胖PCOS患者卵巢颗粒细胞中糖原合成酶激酶-3β(GSK-3β)对糖代谢调控的影响。方法在行IVF/ICSI-ET助孕治疗的患者中,选取10例PCOS患者,根据体质量指数(BMI)分为肥胖组(BMI≥28,n=5)和非肥胖组(18.5<BMI≤23.9,n=5);同期选择月经周期正常的育龄期妇女(18.5<BMI≤23.9,n=5)。取卵日留取并体外培养卵巢颗粒细胞,采用2-脱氧葡萄糖荧光类似物(2-NBDG)检测颗粒细胞葡萄糖摄取能力,己糖激酶(HK)法检测颗粒细胞培养液内葡萄糖(G)、电化学发光法检测颗粒细胞培养液中睾酮(T),激酶活性比色法检测卵巢颗粒细胞内GSK-3β激酶活性,Western Blot法检测丝氨酸9位点发生磷酸化的GSK-3β(p-GSK-3β(Ser9))表达。结果与正常对照组相比,PCOS肥胖组和非肥胖组第48小时细胞培养液内G、T含量均增高(P<0.05);颗粒细胞葡萄糖摄取能力下降(P<0.001);GSK-3β酶活性上升(P<0.05);p GSK-3β(Ser9)的表达下降(P<0.05)。与PCOS非肥胖组相比,PCOS肥胖组第48小时细胞培养液内G、T含量升高(P<0.05);颗粒细胞葡萄糖摄取能力降低(P<0.001);GSK-3β酶活性上升(P<0.05);p-GSK-3β(Ser9)的表达量下降(P<0.05)。结论肥胖影响多囊卵巢综合征患者已经存在的内分泌与糖代谢紊乱。丝氨酸9位点发生磷酸化的GSK-3β可能在肥胖型多囊卵巢综合征的糖代谢异常中发挥重要作用。
Objective：To study the effect of glycogen synthase kinese-3β（GSK-3β）on the glucose metabolism of patient with polycystic ovarian syndrome（PCOS）. Methods：Ten PCOS patients who underwent an IVF/ICSI-ET cycle because of infertility were enrolled in this study. Based on body mass index（BMI）,patients were divided into obese PCOS group（BMI≥28,n=5）and non-obese PCOS group（18.5BMI≤23.9,n=5）. During the same period,5 women in same age with regular menstrual cycles were regarded as non-PCOS group（18.5BMI≤23.9,n=5）. Granulosa cells were collected from patients during IVF/ICSI-ET and in vitro treated for 48 h. The cultured granulosa cells were examined for glucose uptake by 2-NBDG uptake. The level of glucose in cell culture medium was tested by the hexokinase（HK）method. Testosterone level was measured by electrochemiluminescence. GSK-3β activity was represented by kinase activity colorimetric method.Western Blotting was used to measure the protein contents of p-GSK-3β（Ser9）. Results：Compared with non-PCOS group,the levels of glucose and testosterone was higher（P〈0.05）and the granulosa cell glucose uptake decreased（P〈0.001）,while the GSK-3β activity higher（P〈0.05）and the level of phosphorylated GSK-3β protein at site Ser9 was found decreased（P〈0.05）in obese and non-obese PCOS groups. Compared with the obese PCOS group,the glucose and testosterone levels were were higher（P〈0.05）and the granulosa cell glucose uptake decreased（P〈0.001）,while the GSK-3β activity higher（P〈0.05）and the level of phosphorylated GSK-3β protein at site Ser9 was decreased in obese PCOS group（P〈0.05）. Conclusions：Obese patients with polycystic ovary syndrome affect existing endocrine and glucose metabolism. p-GSK-3β（Ser9）may be emerging as important in the PCOS with obesity.
出处
《中国优生与遗传杂志》
2015年第4期118-121,F0004,共5页
Chinese Journal of Birth Health & Heredity
