孕康口服液调节复发性流产小鼠内分泌系统及VEGF信号通路降低流产率的作用和机制

Effect and mechanism of Yunkang oral liquid in regulating endocrine system and VEGF signaling pathway and reducing abortion rate in recurrent abortion mice

研究孕康口服液对Clark经典复发性流产模型小鼠流产率、内分泌系统及VEGF信号通路的影响。采用Clark经典复发性流产模型,成功复制RSA（recurrent sponiaeous abortion）模型小鼠并随机分为模型组,孕康口服液低、中、高3个剂量组,黄体酮组,正常妊娠的小鼠为正常组;从妊娠第1天起,分别给予蒸馏水,孕康口服液9,18,36 m L·kg-1·d-1,黄体酮0.039 g·kg-1·d-1。于妊娠第15天处死小鼠,观察各组小鼠胚胎丢失率,血清雌激素（E2）、孕激素（P）、催乳素（PRL）、黄体生成素（LH）、促卵泡生成素（FSH）水平,子宫蜕膜雌、孕激素受体（ER,PR）、催乳素受体（PRLR）蛋白以及子宫蜕膜RAS,MAPK,VEGF,VEGFR-2基因及蛋白表达。结果显示,孕康中、高剂量、黄体酮能显著降低RSA小鼠的胚胎丢失率;孕康高剂量能显著升高RSA小鼠血清中FSH激素水平;孕康低、中、高剂量均能显著升高RSA小鼠血清中LH,PRL激素水平;孕康中、高剂量升高RSA小鼠血清中P,E2水平;黄体酮仅能显著升高RSA小鼠血清中P,PRL激素水平。Western blot结果显示,孕康低、中、高剂量、黄体酮能显著升高RSA小鼠子宫蜕膜中PRLR,PR,ER表达,且孕康高剂量改善ER的表达强于黄体酮组;Western blot及PCR结果显示孕康中、高剂量、黄体酮均能显著升高RSA小鼠子宫蜕膜组织中RAS,MAPK,VEGF,VEGFR-2基因及蛋白表达。结果说明孕康口服液能有效降低RSA模型小鼠的胚胎丢失率,该作用与升高血清中FSH,LH,PRL,P,E2激素水平,促进小鼠子宫蜕膜中PRLR,PR,ER蛋白表达以及子宫蜕膜中RAS,MAPK,VEGF,VEGFR-2基因及蛋白表达,促进母胎界面血管重铸,改善小鼠子宫内膜容受性,促使蜕膜发育,促进胚泡着床有关。

This experiment focuses on the effect of Yunkang oral liquid on abortion rate,endocrine system and VEGF signal pathway in Clark classical recurrent abortion model mice. RSA mice were randomly divded into model group,low,middle and high-dose groups and progesterone group. The normal pregnancy mice were included into normal group. Since the first day of pregnancy,the normal group and the RSA model group were given the same dose of distilled water,while low,middle and high-dose groups were given Yunkang oral liquid at the dose of 9,18,36 m L·kg1·d-1; progesterone group were given progesterone by 0. 039 g·kg1·d-1. The mice were put to deathat the 15 th day of pregnancy,and the embryo loss rate of each group was observed. Serum estradiol（ E2）,progesterone（ P）,prolactin（ PRL）,luteinizing hormone（ LH）,follicle stimulating hormone（ FSH） level were tested; the protein expressions of estrogen receptor（ ER）,progesterone receptor（ PR）,prolactin receptor（ PRLR） in decidua and RAS,MAPK,VEGF,VEGFR-2 gene and protein expressions in deciduas were studied. The results showed that middle,high dose Yunkang and progesterone could significantly decrease the embryo loss rate of RSA mice. The levels of FSH,LH,PRL,P and E2 in serum in Yunkang and progesterone groups were increased,and the serum levels of FSH,LH,and E2 in Yunkang group were higher than those in progesterone group.Western blot analysis showed that Yunkang oral liquid and progesterone can significantly increase the expressions of PRLR,PR in the uterine decidua of RSA mice,and the expression of ER in Yunkang group was higher than that in progesterone group. Western blot and PCR showed that the Yunkang oral liquid and progesterone can significantly increase RAS,MAPK,VEGF,VEGFR-2 gene and protein expressions in the uterine decidua of RSA mice. The results showed that Yunkang oral liquid can effectively reduce the embryo loss rate of RSA model mice,increase the levels of FSH,LH,PRL,P and E2 in serum,promote the expressions of PRLR,PR,ER protein in decidua and the RAS,MAPK,VEGF,VEGFR-2 gene and protein expressions in the decidua,improve the vascular remodeling of fetal interface,the endometrial receptivty,the development of decidua and the blastocyst implantation.