不同方案超排卵下小鼠子宫内膜种植窗期整合素β3的表达

The expression of integrin beta3 in mice's endometrium during the implantation window based on different ovarian stimulation protocols

目的 以超排卵下小鼠子宫内膜种植窗期整合素 β3的表达为指标 ,比较不同超排卵方案对小鼠子宫内膜容受性的影响。方法 模拟人类超排卵长周期 ,对小鼠进行超促排卵。根据超排卵方案的不同 ,按随机表顺序将 4 0只小鼠分为 4组 ,分别为促性腺激素释放激素拮抗剂 (GnRHant)组、促性腺激素释放激素激动剂 (GnRHa)组、孕马血清促性腺激素 (PMSG)组及对照组 ,每组 10只小鼠。用免疫组织化学和逆转录 聚合酶链反应 (RT PCR)方法分别检测小鼠子宫内膜种植窗期整合素β3蛋白和mRNA的表达。结果 免疫组织化学结果显示 ,与对照组相比GnRHa组小鼠整合素 β3蛋白的表达明显下降 (P =0 0 2 3) ,GnRHant组和PMSG组小鼠整合素 β3蛋白的表达明显降低 (P =0 0 0 1) ;与PMSG组小鼠相比 ,GnRHa组小鼠整合素 β3的表达明显增高 (P =0 0 0 1) ,而GnRHant组整合素 β3的表达差异无显著意义 (P =0 76 8) ;RT PCR结果与免疫组化结果相符。 结论与小鼠自然周期相比 ,3种超排卵方案均使小鼠子宫内膜种植窗期整合素 β3的表达明显下降 ,可能降低了子宫内膜容受性 ;与单一PMSG超排卵方案相比 ,GnRHa辅助超排卵方案能明显改善小鼠子宫内膜的容受性 ;而GnRHant辅助超排卵方案则对小鼠子宫内膜容受性的改善作用不明显。

Objective To compare the uterine receptivity with different ovarian stimulation protocols,the endometrial expression of Integrin beta3 in mice′s during the implantation window underwent different ovarian stimulation were studied. Methods Forty mice were randomly allocated into 4 groups of 10 mice. (1)GnRHant group,gonadotropin-releasing hormone antagonist (GnRHant) was given first for desensitizing the pituitary,then the pregnant mare′s serum gonadotrophin (PMSG) was added for ovarian stimulation. (2)gonadotropin-releasing hormone agonist (GnRHa) group,GnRHa was given first for desensitizing the pituitary,then PMSG was added for ovarian stimulation. (3) PMSG group,injected with PMSG only; and (4) control group,given with saline of the same volume. Human chorionic gonadotropin (HCG) was injected to the mice of the GnRHant,GnRHa,and PMSG groups. Forty-eight hours after the mice of the control group the mice were killed. Their uteri were taken. RT-polymerase chain reaction (RT-PCR) was used to detect the expression of integrin beta3 mRNA. Immunohistochemistry (SP method) was used to locate and semiquantitatively measure the integrin beta3 in the endometrium during implantation window. Results (1)Immunohistochemistry: showed that the staining intensity of integrin beta3 was 3.74±0.15 in GnRHa group,3.22±0.19 in the GnRHant group,and 3.24±0.18 in the PMSG group,all significantly lower than that in the control group (3.90±0.11,P =0.023,0.001,and 0.001).with a significant difference between the GnRHa group and the PMSG group ( P =0.001) and without a significant difference between the .GnRHant group and PMSG groups ( P =0.768).(2)RT-PCR showed that the relative quantity of integrin β3 mRNA expression was 1.14±0.16 in the GnRHa group,0.76±0.33 in the GnRHant group,and 0.73±0.26 in the PMSG group,all significantly lower than that in the control group (1.4±0.3,P =0.045,0.001,and 0.001). with a significant difference between the GnRHant and PMSG groups ( P =0.001) and without a significant difference between the GnRHant and PMSG groups ( P = 0.857). Conclusion All of the ovarian stimulation protocols do harm to the mice′s uterine receptivity. Ovarian stimulation combining GnRHa protocol in mice is better than PMSG alone protocol,because it can improve the uterine receptivity. Ovarian stimulation combining GnRHant protocol in mice decrease the uterine receptivity as well as the PMSG alone protocol.