补肾调经方在种植窗期经STAT3信号通路改善超促排卵小鼠胚胎着床的作用及机制

Effects and mechanisms of Bushen Tiaojing Fang on the embryo implantation via STAT3 signaling pathway during implantation window in superovulated mouse

目的观察补肾调经方对控制性超促排卵(COH)小鼠种植窗期子宫内膜信号转导和转录活化因子3(STAT3)信号通路的影响,探讨其改善胚胎着床的作用及机制。方法昆明小鼠60只,随机分为模型组、补肾调经方组和正常组,每组20只。妊娠第4天每组处死10只小鼠,放射免疫法检测血清雌二醇(E_2)、孕酮(P)水平,Western blot法检测子宫内膜白血病抑制因子(LIF)、磷酸化信号转导和转录活化因子3(P-STAT3)蛋白的含量,免疫组化法检测子宫内膜瘦素及瘦素受体(OB-R)的表达。妊娠第8天处死各组剩余小鼠,观察平均胚胎着床数。结果与正常组比较,模型组小鼠血清E_2降低(P<0.05),P升高(P<0.01);子宫内膜LIF、P-STAT3的含量下降(P<0.01),内膜上皮细胞瘦素及OB-R蛋白的表达降低(P<0.01);平均胚胎着床数减少(P<0.01)。与模型组比较,补肾调经方组小鼠血清P降低(P<0.05);子宫内膜LIF、P-STAT3蛋白的含量升高(P<0.01),内膜上皮细胞瘦素表达增加(P<0.05);平均胚胎着床数增多(P<0.01)。结论补肾调经方可改善COH小鼠胚胎着床,其机制可能与调控种植窗期子宫内膜STAT3信号通路相关蛋白有关。

Objective To observe the effects of Bushen Tiaojing Fang（ BSTJF,kidney-tonifying menstruation-regulating formula） on endometrium of controlled ovarian hyperstimulated（ COH） mice through signal transducer and activator of transcription 3（ STAT3）,and to explore mechanisms of BSTJF for improving embryo implantation. Methods Sixty Kunming mice were randomly divided into the model group,BSTJF group and normal group（ n = 20 in each group）. On the fourth day of pregnancy,10 mice from each group were sacrificed. The levels of serum estradiol（ E_2） and progesterone（ P） were measured by using radioimmunoassay. The expressions of endometrial leukemia inhibitory factor（ LIF）,phosphorylated STAT3（ P-STAT3） protein were evaluated with Western blot assay; Leptin and leptin receptor（ OB-R） were detected with immunohistochemistry. At eighth day when all mice were then sacrificed,the average number of blastocysts was counted. Results Compared with the normal group,serum E_2 level in the model group was decreased（ P 〈 0. 05）,and P was increased（ P 〈 0. 01）; the contents of LIF and P-STAT3 protein in the endometrium decreased（ P 〈 0. 01）; the expressions of Leptin and OB-R in epithelial cells decreased（ P 〈 0. 01）; the average number of implanted embryos was also decreased（ P 〈 0. 01）. Compared with the model group,serum P decreased in the BSTJF group（ P 〈 0. 05）; the contents of LIF and P-STAT3 protein in endometrium increased（ P 〈 0. 01）,the expression of Leptin protein in endometrial epithelial cells increased（ P 〈 0. 05）. The average number of blastocysts was increased（ P 〈 0. 01）. Conclusion BSTJF can improve the embryo implantation of COH mice. Its mechanism might be related to regulation of relative proteins in STAT3 signal pathway in the endometrium during implantation window.