GSK-3β抑制剂对体外培养多囊卵巢综合征患者卵巢颗粒细胞睾酮分泌过多的影响

Effect of GSK3 β inhibitor on Testosterone Excess within in Vitro Cultured Ovarian Granulosa Cells in PCOS Patients

目的探讨糖原合成酶激酶-3(GSK-3)抑制剂SB216763改善多囊卵巢综合征(PCOS)患者卵巢颗粒细胞睾酮(T)分泌异常的作用机制。方法将行体外受精-胚胎移植(IVF-ET)治疗的20例不孕妇女按发病原因分为三组,其中不孕原因为输卵管及男方因素的非PCOS患者8例为对照组,PCOS不孕患者12例平均分为PCOS组和GSK-3β抑制剂组,每组6例。对照组及PCOS组患者的卵巢颗粒细胞进行体外培养48 h,利用放射免疫法检测细胞上清液中的雌二醇(E2)、孕酮(P)、T的水平。GSK-3β抑制剂组用浓度为5、10、25、50、100μmol/L的GSK-3β抑制剂SB216763对体外培养的PCOS患者卵巢颗粒细胞进行干预,后继续培养48 h,检测细胞上清液中的E2、P、T的水平。结果 (1)PCOS组与对照组患者相比其卵巢颗粒细胞中T分泌明显升高(P<0.01),而P和E2的表达变化不大(P>0.05)。(2)GSK-3β抑制剂组PCOS患者卵巢颗粒细胞给予不同浓度的GSK-3β抑制剂SB216763作用后,与PCOS组相比,T平明显下降(P<0.05)。(3)GSK-3β抑制剂SB216763浓度为5μmol/L时对PCOS患者卵巢颗粒细胞T分泌的抑制作用最明显(P<0.01)。结论 PCOS患者卵巢颗粒细胞T分泌异常增高,GSK-3β抑制剂SB216763能够抑制其T的过多分泌。

Objective To explore the function mechanism of( glycogen synthase kinase) GSK- 3 inhibitor,SB216763 on testosterone( T) excess in ovarian granulosa cells( GC) in the patients of polycystic ovarian syndrome( PCOS). Methods Twenty infertility women with in vitro IVF- ET were divided into three groups according to causative factors,in which,8 cases of non- PCOS due to the factors of fallopian and husband’s side were in the control group and 12 cases of PCOS were equally subdivided into a PCOS group and a GSK- 3β inhibitor group,6 cases in each one. In the control group and PCOS group,GCs were cultured in vitro for 48 h. RIA was used to detect the levels of estradio( E2),progesterone( P) and T in the supernatant.In the GSK- 3β inhibitor group,GSK- 3β inhibitor,SB216763,at 5,10,25,50 and 600 μmol / L was used in the intervention of in vitro cultured GCs in PCOS patients. After cultured continuously for 48 h,the levels of E2,P and T in the supernatant were detected. Results 1. T secretion in the GCs was increased apparently in PCOS group as compared with that in the control group( P < 0. 01),and the expressions of P and E2 did not change a lot( P > 0. 05). 2. In the GSK- 3β inhibitor group,after the intervention with GSK- 3β inhibitor,SB216763 of different concentrations,compared with PCOS group,T level was reduced apparently( P <0. 05). 3. GSK- 3β inhibitor,SB216763 at 5 μmol / L displayed the most obvious inhibition on T secretion inPCOS patients( P < 0. 01). Conclusion T secretion in GCs is increased abnormally in PCOS patients. GSK- 3β inhibitor,SB216763 inhibits T excess.