寿胎丸提高控制性超排卵小鼠子宫内膜容受性的机制研究

Study on Mechanism of Shoutai Pill(寿胎丸)Improving Endometrial Receptivity in COH Mice

目的通过研究寿胎丸对控制性超排卵(Controlled ovarian hyperstimulation,COH)小鼠子宫内膜整合素αv(Integrinαv,ITGαv)、整合素α5(Integrinα5,ITGα5)、整合素β3(Integrinβ3,ITGβ3)及白血病抑制因子(Leukemia inhibitory factor,LIF)的影响,探讨寿胎丸提高COH子宫内膜容受性的作用机制。方法采用促性腺激素释放激素激动剂(Gonadotropin-releasing hormone agonist,GnRHa)+孕马血清促性腺激素(Pregnant mare serum gonadotropin,PMSG)+绒毛膜促性腺激素(Chorionic gonadotropin,HCG)建立COH小鼠模型,将其分为COH模型组、寿胎丸高、中、低剂量组、阿司匹林对照组以及正常对照组,在造模的同时,寿胎丸高、中、低剂量组分别按6、3、1 g/kg寿胎丸灌胃;阿司匹林对照组按0.2 g/kg阿司匹林灌胃;COH模型组、正常对照组以等体积生理盐水灌胃,每天1次,连用11 d。采用免疫组化技术检测子宫内膜ITGαv、ITGα5、ITGβ3及LIF表达情况,采用逆转录-聚合酶链反应(Reverse transcription-polymerase chain reaction,RT-PCR)技术检测子宫组织ITGαv、ITGα5、ITGβ3及LIFmRNA表达水平。结果与正常对照组相比较,COH模型组子宫内膜ITGαv、ITGα5、ITGβ3、LIF蛋白阳性染色的积分光密度值(Integral optical density,IOD)及区域(Area)明显减少(P<0.05),子宫ITGαv、ITGα5、ITGβ3、LIF mRNA水平明显降低(P<0.05)。与COH模型组比较,寿胎丸高、中、低剂量组子宫内膜ITGαv、ITGα5、ITGβ3及LIF蛋白阳性染色的IOD及Area明显增加,子宫ITGαv、ITGα5、ITGβ3及LIFmRNA水平明显升高(P<0.05)。结论寿胎丸提高COH模型小鼠子宫内膜容受性可能与调节子宫内膜ITGαv、ITGα5、ITGβ3、LIF蛋白含量及mRNA水平有关。

Objective By investigating the effect of Shoutai Pill(寿胎丸)on endometrial integrins[Integrinαv(ITGαv),Integrinα5(ITGα5),Integrinβ3(ITGβ3)]and leukemia inhibitory factor(LIF)in mice with controlled superovulation(COH)to explore the mechanism of Shoutai Pill in improving endometrial receptivity of COH.Methods COH mice model was established by using gonadotropin-releasing hormone agonist(GnRHa)+pregnant mare serum gonadotropin(PMSG)+chorionic gonadotropin(HCG)and the mice were divided into 6 groups:the model group,the high,medium and low dose groups of Shoutai Pill,the aspirin control group,as well as the normal control group.While mice modeling,the high,medium and low dose groups of Shoutai Pill were given Shoutai Pill by gavage with the dose of 6 g/kg,3 g/kg and 1 g/kg,respectively.The aspirin control group was given 0.2 g/kg aspirin by gavage.The COH model group and the normal control group were gavaged with an equal volume of 0.9%normal saline,once a day for 11 days.The expressions of ITGαv,ITGβ3,ITGα5 and LIF protein in endometrium were detected by immunohistochemistry method,and the mRNA expression levels of ITGαv,ITGβ3,ITGα5 and LIF in endometrial tissues were detected by RT-PCR.Results Compared with thsoe of the normal control group,the integral optical density(IOD)and area of endometrial ITGαv,ITGα5,ITGβ3 and LIF protein positive staining were significantly reduced(P<0.05)and endometrial ITGαv,ITGα5,ITGβ3 and LIF mRNA levels were significantly decreased(P<0.05)in the COH model group.Compared with those of the COH model group,the IOD and area of endometrial ITGαv,ITGβ3,ITGα5 and LIF protein positive staining as well as the endometrial ITGαv,ITGβ3,ITGα5 and LIF mRNA levels were significantly increased(P<0.05)in the high,medium and low dose groups of Shoutai Pill.Conclusion The improvement of endometrial receptivity by Shoutai Pill in COH mice may be related to the regulation of endometrial LIF,ITGαv,ITGβ3 and ITGα5 protein contents and mRNA levels.