泰山磐石散对PCOS促排卵大鼠早期妊娠丢失的干预作用
Intervention Effect of Taishan Panshi San on Early Pregnancy Loss in PCOS Rats with Ovulation Induction
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摘要: 目的 探讨多囊卵巢综合征(PCOS)促排卵大鼠发生早期妊娠丢失的可能机制及泰山磐石散对妊娠丢失的干预作用。方法 将正常大鼠与雄鼠合笼,选取妊娠大鼠作为A组。将脱氢表雄酮(DHEA)诱导PCOS模型大鼠经克罗米芬(CC)促排卵后与雄鼠合笼,建立PCOS促排卵妊娠大鼠模型,并随机分为:PCOS促排妊娠+生理盐水组(B组),PCOS促排妊娠+泰山磐石散组(C组)。B组按照妊娠d3、d4、d5,分别设为B3组、B4组、B5组。以RT-PCR、Western blot分别检测各组妊娠大鼠子宫内膜同源框蛋白(HOXA10)、整合素ανβ3、同源结构域基因(EMX2)、白血病抑制因子(LIF)的mRNA及蛋白的表达。结果 在PCOS促排妊娠B组中:B4组HOXA10 mRNA表达明显高于B5组、B3组,差异有显著性(P<0.05~0.01),整合素ανβ3 mRNA表达明显高于B5组、B3组,有显著性差异(P<0.01),而EMX2-mRNA表达明显低于B5组,差异有显著性(P<0.05);B3组LIF mRNA的表达最高,与B4组组间差异显著(P<0.05)。在种植窗期:B5组HOXA10、整合素ανβ3、LIF的mRNA表达均明显低于A组、C组,差异有显著性(P<0.05~0.01);A组EMX2的mRNA表达明显低于B5组、C组,差异有显著性(P<0.01);B5组HOXA10、整合素ανβ3、LIF的蛋白表达均明显低于A组、C组,而EMX2明显高于A组、C组,有显著差异(P<0.01)。结论 PCOS促排大鼠发生早期妊娠丢失可能与种植窗期子宫内膜HOXA10、整合素ανβ3、LIF的低表达、EMX2的高表达有关;也可能与种植窗前期子宫内膜HOXA10、整合素ανβ3、LIF、EMX2表达量异常有关。泰山磐石散可能通过上调种植窗期子宫内膜HOXA10、整合素ανβ3、LIF,下调EMX2的表达,提高子宫内膜容受性,减少早期妊娠丢失。Abstract: OBJECTIVE To investigate the possible mechanism of early pregnancy loss in PCOS rats with ovulation induction and the intervention effect of Taishan Panshi San on pregnancy loss. METHODS To select normal pregnant rats mated with male rats as group A. The PCOS rats model by dehydroepiandrosterone(DHEA) was stimulated ovulation by clomiphene(CC),then mated with male rats to establish the rat model of pregnancy.They were randomly divided into two groups:treated by normal saline (group B) and by Taishan Panshi San (group C). According to the pregnancy third days, fourth days, fifth days, group B was divided into group B3, group B4, group B5. To detect the mRNA and protein expression of endometrium HOXA10, integrinαvβ3, EMX2, LIF by RT-PCR and Western blot in each group. RESULTS In group B of PCOS Pregnant rats with ovulation induction:The expression of mRNA HOXA10 in Group B4 was higher than group B3 and group B5(P<0.05~0.01),the expression of mRNA integrinαvβ3 was significantly higher than group B3 and group B5(P<0.01),but the expression of mRNA EMX2 was lower than group B5(P<0.05);the expression of mRNA LIF was highest in group B3, higher than group B4(P<0.05)。In the planting window period:the mRNA expression of endometrium HOXA10, integrinαvβ3, LIF in Group B5 were significantly lower than Group A and Group C (P<0.05~0.01);the expression of endometrium EMX2 in Group A was significantly lower than Group B5 and Group C (P<0.01);the protein expression of endometrium HOXA10, integrinαvβ3, LIF in Group B5 were significantly lower than Group A and Group C ,but the protein expression of EMX2 higher than Group A and Group C (P<0.01). CONCLUSION PCOS rats with ovulation induction occurred early pregnancy loss may be related to the low expression of endometrial HOXA10,integrinαvβ3, LIF and the high expression of EMX2 in the implantation window phase. This may also be related to the abnomal expression of HOXA10, integrinαvβ3, LIF, EMX2 before the planting window.Taishan Panshi San improved the endometrial receptivity and reduced the early pregnancy loss,probably by raising the expressions of endometrium HOXA10, integrinαvβ3, LIF and decreasing the expressions of EMX2 in the implantation window phase.