中药助孕方对促排卵和胚泡着床障碍小鼠子宫内膜容受性的影响

Effects of Zhuyun Recipe on the Endometrial Receptivity in Mice with Blastocyst Implantation Dysfunction and Ovulation Stimulation

目的研究中药助孕方对促排卵(OS)和胚泡着床障碍(BID)小鼠子宫内膜容受性的影响及其潜在机制。方法 200只正常雌性昆明小鼠,随机分为A组(对照组)、B组(OS)、C组(OS加中药助孕方)、D组(BID)、E组(BID加中药助孕方)、F组(中药助孕方)。OS组腹腔内注射孕马血清促性腺素和人绒毛膜促性腺素(HCG),BID组于妊娠第4天9:00皮下注射米非司酮。3个中药组于妊娠第1-4天8:00灌胃中药1.5mL/100g。妊娠第4天21:00每组取出8只子宫标本固定,运用免疫组化的方法测定标本中白血病抑制因子(LIF)和整合素β3的表达水平。妊娠第8天21:30处死各组妊娠小鼠,取出子宫,计数胚泡数。结果 B组和D组的妊娠率(6.67%,1/15、18.75%,3/16),LIF蛋白OD值(0.18±0.02、0.23±0.02),整合素β3蛋白OD值(0.20±0.05、0.19±0.02),与A组比较差异有统计学意义(P<0.01)。C组和E组的妊娠率(54.55%,12/22、65.22%,15/23),LIF蛋白OD值(0.37±0.09、0.39±0.02),整合素β3蛋白OD值(0.34±0.04、0.38±0.08)分别与B组和D组比较差异有统计学意义(P<0.05)。结论 OS和BID对小鼠子宫内膜容受性有负面影响,阻碍胚泡植入。中药助孕方可以通过上调LIF和整合素β3的表达来改善子宫内膜容受性,提高妊娠率。

Objective To study the effects and underlying mechanisms of Zhuyun Recipe （ZR） on the endometrial receptivity in ovarian stimulation （OS） and blastocyst implantation dysfunction （BID） mice. MethodsTotally 200 normal female Kunming mice were randomly divided into 6 groups, i.e., the control group （Group A）, the OS group （Group B）, the OS＋ZR group （Group C）, the BID group （Group D）, the BID＋ZR group （Group E）, and the ZR group （Group F）. The pregnant mare′s serum gonadotrophin （PMSG） and human chorionic gonadotrophin （HCG） were intraperitoneally injected to mice in Group B. Mifepristone was subcutaneously injected to mice in Group D at 9：00 am on the 4th gestation day. Corresponding medications were given to mice in Group C, E, and F at 1.5 mL/100 g by gastrogavage at 8：00 am from the first to the 4th gestation day. Eight uterus samples were collected at 9：00 pm on the 4th gestation day and fixed. The expression levels of leukemia inhibitory factor （LIF） and integrin β3 were detected using immunohistochemical assay. The pregnant mice were sacrificed at 9：30 pm on the 8th gestation day, and their uterus were taken out. The number of blastocysts was counted. ResultsCompared with Group A, the pregnant rate was 6.67% （1/15 cases） in Group B and 18.75% （3/16 cases） in Group D, the mean OD value of LIF was 0.18±0.02 in Group B and 0.23±0.02 in Group D, and the mean OD value of integrin β3 was 0.20±0.05 in Group B and 0.19±0.02 in Group D, showing statistical difference （P0.01）. The pregnant rate was 54.55% （12/22 cases） in Group C and 65.22% （15/23 cases） in Group E, the mean OD value of LIF was 0.37±0.09 in Group C and 0.39±0.02 in Group E, and the mean OD value of integrin β3 was 0.34±0.04 in Group C and 0.38±0.08 in Group E, showing statistical difference when compared with those of Group B and Group D respectively （P0.05）. Conclusions OS and BID had negative effects on the endometrial receptivity and hindered the blastocyst implantation. ZR could improve the uterine receptivity and elevate the pregnant rate by up-regulating the expressions of endometrial LIF and integrin β3.