EGF通过PI3K/AKT通路介导小鼠卵巢颗粒细胞增殖

目的探讨表皮生长因子（EGF）介导,经PI3K/AKT信号通路调控小鼠卵巢颗粒细胞增殖的作用与机制。方法取分离纯化的小鼠卵巢颗粒细胞,添加10ng·mL^-1的EGF对其培养24h后,然后再添加20μM浓度的P13K抑制剂LY294002处理细胞72h,利用CCK-8检测颗粒细胞增殖情况;应用Real-time PCR法检测AKT mRNA的表达;应用Western blot法检测体外培养小鼠颗粒细胞中总AKT（t-AKT）及磷酸化AKT Thr308位点（p-AKTThr308）蛋白的表达情况。结果 （1）10ng·mL^-1的EGF对小鼠颗粒细胞增殖效应明显,能够显著提高AKT基因在颗粒细胞中的表达（P〈0.05）,能显著性促进t-AKT及p-AKTThr308蛋白的表达（P〈0.05）。（2）PI3K特异性抑制剂LY294002明显抑制EGF促进颗粒细胞增殖的效应（P〈0.05）。结论EGF能活化小鼠颗粒细胞的PI3K/AKT信号通路,促进颗粒细胞生长、增殖。

EGF Promotes AKT Expression and Phosphorylation in Mouse Ovarian Granulosa Cells Through the Activation of PI3K/AKT

Objective To explore the effects of epidermal growth factor （EGF）on AKT expression and phos- phorylation. Methods Mouse ovarian granulosa cells were cultured in TCM199 medium supplemented with 10 ng · mL^-1 EGF, and to investigate the effects of PI3 K inhibitor, LY294002,on suppression of AKT and regulation of granulosa cells in vitro. CCK -8 was used to study the proliferation of ovarian granulosa cells. The AKT expression and Thr308 - phosphorylation level in granulosa cells were detected by Real -time PCR and Western blot for 24 hours after EGF treatment. Results It was found that EGF significantly improved the proliferation of ovarian granulosa cells. The AKT mRNA and AKT protein expression of granulosa cells showed increase significantly after treatment of 10ng · mL^-1 EGF（ P 〈 0.05 ） , the Thr308 - phosphorylation level of AKT was increased in EGF treated cells relative to the control cells （ P 〈 0.05 ）. Conclusion The AKT pro- tein and Thr308 -phosphorylation level were increased significantly after EGF treatment in Mouse ovarian granulosa cells in vitro culture. The protective effect of EGF against apoptosis is known to occur through the activation of PI3K/AKT, LY294002 may represent a novel and effective target of intervention strategies to sup- press PI3K/AKT expression.