电针对控制性超排卵小鼠子宫内膜磷酸化胰岛素样生长因子-1受体表达及其信号转导通路的调控

Regulation effect of electro-acupuncture on expression of endometrium phosphorylation insulin-like growth factor-1 receptor and its signal transduction pathways in mice with controlled ovarian hyperstimulation

目的观察电针对控制性超排卵(COH)小鼠子宫内膜胰岛素样生长因子-1受体(IGF-1R)表达及丝裂原激活蛋白激酶(MAPK)、磷酯酰肌醇-3-羟激酶(PI3K)信号转导通路的影响,探讨电针改善胚胎着床的作用机制。方法将昆明纯种性成熟雌性小鼠随机分成自然周期组、COH组、电针组、电针+BMS-536924组、电针+LY-294002组、电针+PD-98059组(其中BMS-536924、LY-294002、PD-98059为IGF-1R、PI3K、MAPK阻断剂的名称),每组20只,每组又分为供体鼠和受体鼠。采用控制性超促排卵长方案制备动物模型,并于注射绒毛膜促性腺素(HCG)日取关元、中极、三阴交行电针治疗,1次/d,至取材停止针刺。取供体鼠受精卵体外培养后移植至同组同日见阴栓的受体鼠子宫内,观察临床妊娠率和胚胎着床位点数,用Western blot法检测子宫内膜IGF-1R、磷酸化胰岛素样生长因子-1受体(p-IGF-1R)、磷酸化蛋白激酶B(p-Akt)、磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)的表达。结果 COH组妊娠率、平均着床位点数和子宫内膜IGF-1R、p-IGF-1R、p-Akt、p-ERK1/2的表达均较自然周期组低(P<0.05或0.01),电针组、电针+LY-294002组、电针+PD-98059组各项指标均较COH组显著性增加(P<0.05或0.01),但电针+BMS-536924组妊娠率、平均着床位点数未见明显改善,且p-IGF-1R表达显著性低于自然周期组(P<0.01)。电针+BMS-536924组、电针+LY-294002组、电针+PD-98059组相比较,电针+PD-98059组p-Akt蛋白和电针+LY-294002组p-ERK1/2蛋白表达较高,其差异具有统计学意义(P<0.01)。结论电针改善COH小鼠胚胎着床可能与上调p-IGF-1R表达及PI3K/Akt、MAPK/ERK1/2信号转导通路有关。

Objective To observe the influence of electro-acupuncture（ EA） on expression of endometrium phosphorylation insulin-like growth factor-1 receptor（ p-IGF-1R）,and signal transduction pathways of MAPK and PI3 K in mice with controlled ovarian hyperstimulation（ COH）,and discuss the mechanism of EA improving embryo implantation. Methods Female Kunming mice were randomly divided into natural cycle group,COH group,EA group,EA ＋ BMS-536924 group,EA ＋ LY-294002 group and EA ＋ PD-98059 group（ each n = 20）,and mice were divide again into donor mice and recipient mice. The model was established by applying controlled hyperstimulation program,and given EA in Guanyuan（ RN3）,Zhongji（ RN4） and Sanyinjiao（ SP6） at the day of injecting human chorionic gonadotropin（ HCG） once a day until stopping collecting samples. The fertilized ovums were collected from donor mice and cultured in vitro,and then transplanted into the uterus of recipient mice. The clinical pregnancy rate and site number of embryo implantation were observed. The expressions of endometrial IGF-1R,p-IGF-1R,phosphorylation protein kinase（ p-Akt） and phosphorylation extracellular regulated protein kinases1/2（ p-ERK1/2） were detected by using Western Blot. Results The clinical pregnancy rate,site number of embryo implantation and expressions of endometrial IGF-1R,p-IGF-1R,p-Akt and p-ERK1/2 were all lower in COH group than those in natural cycle group（ P 〈 0. 05 or P 〈 0. 01）. All indexes were higher in EA group,EA ＋ LY-294002 group and EA ＋ PD-98059 group than those in COH group（ P 〈 0. 05 or P 〈 0. 01）. But the clinical pregnancy rate,site number of embryo implantation had no significant improvement in EA ＋ BMS-536924 group,and expression of p-IGF-1R was significantly lower than that in natural cycle group（ P 〈 0. 01）. The comparison among EA ＋ BMS-536924 group,EA ＋ LY-294002 group and EA ＋ PD-98059 group showed that the expression of p-Akt was higher in EA ＋ PD-98059 group, and expression of p-ERK1/2 was higher in EA ＋ LY-294002 group（ P 〈 0. 01）.Conclusion Embryo implantation can be improved by EA in COH mice,which may be related to EA can up-regulate the expression of p-IGF-1R and signal transduction pathways of PI3K/Akt and MAPK/ERK1/2.