Comparative Study Of Intervention About Bushenantai Recipe And Its Two Components On Embryonic Implantation Of Controlled Ovarian Hyperstimulation

Objective:To investigate the roles of Bushenantai recipe and its two components during implantation in mice of COH (controlled ovarian hyperstimulation), and definite selective or cooperative effects between Bushen and Huoxue components; we observe the effects of the recipe and its two components on pregnancy rate and the expression of HB-EGF (heparin-binding epidermal growth factor-like growth factor) and EGFR (epidermal growth factor receptor) in mice, furthermore, provide experimental bases of kidney-reinforcement and blood-activation for the treatment of infertility.Method:All mice were randomly divided into normal group, model group, Bushenantai group, Bushen group and Huoxue group. Model and TCM groups were intraperitoneal injected with PMSG 10IU at 5pm, then mated with fertile males (day1=vaginal plug) after intraperitoneal injected with HCG 10IU 48 hours later. Normal group would be mated without COH. And observed vaginal plug rate at pregnancy day 1(Pd1), pregnancy rate and the number of implantation sites at Pd6. Uterus were collected on Pd5 and 6. Detected the expression of HB-EGF mRNA by RT-PCR, expression of EGFR were analyzed by immunohistochemistry. Result:1. Comparison of vaginal plug,pregnancy rates and number of implantation sites The vaginal plug and pregnancy rates in model group were significantly lower than normal, but the number of implantation sites showed the conflicting results. The vaginal plug rate and pregnancy rates in every TCM groups were higher significantly than model group respectively; Bushenantai group were higher than Bushen and Huoxue group, but the number of implantation sites conflicted. In Huoxue group, there were some differences. The vaginal plug rate was higher than Bushen group, and the number of implantation sites showed the conflicting results, but there is no statistical difference in pregnancy rates between the two groups.2. Comparison of morphologyBlastocysts of normal group were well-distributed and line up well in mouse uterus, evans blue dye was homogeneous, but the number of implantation sites were fewer than model group. Blastocysts of model group were uneven distribution and partly inosculated, evans blue uneven dyed. Blastocysts of TCM group showed growing better than model group, which in Bushenantai group are catenuliform, their shape and distribution both better than component groups. Blastocysts in Huoxue group developed better than Bushen group.3. Comparison of RT-PCR resultsThe expression of HB-EGF mRNA in the mice endometrium of COH was significantly lower than normal group. Bushenantai recipe and its Bushen, Huoxue components could improve the expression of HB-EGF in model mice. Respectively, the expression of HB-EGF mRNA in Huoxue group higher than Bushen group, but lower than Bushenantai group; in general terms, the expression of HB-EGF mRNA on Pd6 was lower than Pd5.4. Comparison of Immunohistochemistry EGFR were expressed on the endometrial glands and stromal cells during implantation. Immunohistochemistry observation suggests that there was no remarkably difference on EGFR spatial distribution in the five groups. Normal groups had compact glands and EGFR expression was rich in nuclear of glandular epithelium cell, meanwhile in cell cytoplasm in mice endometrium; the endometrial glands were sparse in model group and EGFR expression was less in nuclear of glandular epithelium cell, meanwhile EGFR expression was deficient in endometrial stromal cell. EGFR in TCM groups expressed stronger than model groups; the expression of EGFR in Bushen group, Huoxue group, Bushenantai group were gradually increased.Conclusion:Bushenantai recipe and its Bushen,Huoxue components could improve the vaginal plug rates, pregnancy rates and normalize the Blastocysts growth, and up-regulated the expression of HB-EGF and EGFR; Bushen and Huoxue components had some cooperated effects in embryo implantation.