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目的:通过研究P38蛋白激酶抑制剂SB203580对多囊卵巢综合征(PCOS)患者卵巢颗粒细胞PI3K通路的影响,探讨PCOS胰岛素抵抗发生的受体后机制。方法:收集在我院行体外受精/卵细胞内单精子注射-胚胎移植(IVF/ICSI-ET)助孕并伴有PCOS不孕患者的卵泡黄素化颗粒细胞进行体外培养(n=10),将每份细胞样本分种于两个培养瓶,一份采用P38丝裂原活化的蛋白激酶信号转导通道(MAPK)抑制剂SB203580处理(PCOS+IN组),一份未加抑制剂(PCOS组)。另收集行IVF/ICSI-ET助孕的非PCOS患者的卵泡黄素化颗粒细胞作为对照组(N-PCOS组,n=10)。Western blot法检测各组颗粒细胞中p-P38MAPK、磷酸化蛋白激酶B(p-Akt)/Akt以及葡萄糖转运蛋白-4(GLUT-4)表达。结果:PCOS组的p-P38MAPK蛋白表达显著高于N-PCOS组,p-Akt/Akt、GLUT-4蛋白表达量较N-PCOS组显著降低(P<0.05);SB203580处理后p-P38MAPK蛋白表达明显降低;p-Akt/Akt、GLUT-4蛋白表达量仍较N-PCOS组低,但较SB203580未处理组表达量高(P<0.05)。结论:PCOS患者的卵巢颗粒细胞中P38/MAPK通路活性升高,磷脂酰肌醇-3激酶途径/Akt(PI3K/Akt)途径活性降低;卵巢颗粒细胞中P38/MAPK途径与PI3K/Akt途径存在相互联系,P38/MAPK通路阻滞剂SB203580可能通过阻断P38MAPK途径的表达,从而提高卵巢颗粒细胞PI3K/Akt通路的活性,提高GLUT-4表达。
Objective: To investigate the effect of P38 protein kinase inhibitor SB203580 on PI3K pathway in ovarian granulosa cells in patients with polycystic ovary syndrome (PCOS) and to explore the mechanism of PCOS after insulin resistance. Methods: The luteinizing granulosa cells (IVF / ICSI-ET) and follicular luteinizing granulosa cells (PCNA) infertile women with PCOS infertility were collected for in vitro culture (n = 10) The cells were subcultured in two flasks, one was treated with P38 mitogen-activated protein kinase signal transduction channel (MAPK) inhibitor SB203580 (PCOS + IN group) and one without inhibitor (PCOS group) . In addition, follicular luteinized granulosa cells from non-PCOS patients who underwent IVF / ICSI-ET were collected as control group (n = 10 in N-PCOS group). Western blot was used to detect the expression of p-P38MAPK, p-Akt / Akt and glucose transporter-4 (GLUT-4) in each group. Results: The protein expression of p-P38MAPK in PCOS group was significantly higher than that in N-PCOS group (P <0.05). The expression of p-Akt / Akt and GLUT-4 protein in PCOS group was significantly lower than that in N-PCOS group The expression of p-Akt / Akt, GLUT-4 protein was still lower than that of N-PCOS group, but higher than that of SB203580 group (P <0.05). Conclusions: The activity of P38 / MAPK pathway in ovarian granulosa cells in PCOS patients is increased, and the activity of phosphatidylinositol 3 - kinase pathway / Akt (PI3K / Akt) pathway is decreased. P38 / MAPK pathway and PI3K / Akt pathway exist in ovarian granulosa cells P38 / MAPK pathway blocker SB203580 may increase the activity of PI3K / Akt pathway and increase the expression of GLUT-4 in ovarian granulosa cells by blocking the expression of P38MAPK pathway.