| Endometriosis(endometriosis,EM,)refers to a kind of estrogen dependent disease,which the growth function of endometrial tissue outside the uterine cavity.Endometriosis is a common disease and frequently encountered disease in women of child-bearing period,causing pain and infertility.The incidence rate of EM is increasing and tending to be younger.Endometriosis associated infertility is as high as 40-60%,and 40%-50%in patients with infertility also is endometriosis.At present,the surgical treatment is the first basic measures for EM associated infertility.With the development of assisted reproductive technology(ART)and clinical application,ART has already become the most ideal assisted reproduction methods and treatment for EM associated infertility.But in IVF-ET,the embryo transfer rate was 85%and the pregnancy rate is only 20%.Therefore,how to improve the pregnancy outcomes of EM associated infertility in IVF-ET is a hot issues.This study will observe the quality of the blastocyst,endometrial receptivity and pregnancy outcome of the model mice through IVF-ET under the intervention of Huo Xuexiaoyi Fang.Discuss possible mechanism of traditional Chinese medicine to promote EM associated infertility pregnancy rate,lay a foundation for traditional Chinese medicine into the field of assisted reproduction,for the patients with EM associated infertility provide better treatments.Objectives:1)To establish the endometriosis mice model and to observe the endometriosis effects of reproductive ability.on model mice2)To observe and select the best time site of implantation period on pinopodes scanning electron microscopy on the endometriosis mice model with pregnancy low function.3)To evaluate Huo Xuexiaoyi Fang the effect of in vitro fertilization and embryo transplantation pregnancy pregnancy outcome assessment on endometriosis model in mice.Methods:1)Through the method of abdominal cavity and subcutaneous injections to establish the endometriosis mice model he endometriosis effects of reproductive ability.on model mice compared with Sham-operated group and the blank group.2)Make the endometriosis mice model with pregnancy low function with infertility ligation mice in cages by2:1,Oh was defined as the day of vaginal plug at 8:00.After examining vaginal plug,the expression of endometrial pinopodes was observed with scanning electron microscopy at 80h,84h,85h,85.5h,86.5h,87h,88h,91h,95h and no false pregnancy.3)By the method of IVF-ET,to observe the quantity of the ovarian stimulation and the rate of fertilization,two-cell stage,four cell stage and blastulation among three groups.Observe the influence of the endometriosis mice with pregnancy low function on the quality of embryos.To explore the influence of Huo Xuexiaoyi Fang on mice embryonic development of in vitro fertilization and embery transplantation.4)According to the different conditions,including Traditional Chinese Medition group(TCM),model group and sham operation group,the embryonics and pseudo pregnancy mouse are divided into nine groups in vitro fertilization and embryo transplantation.A week after transplantation,we observe the pregnancy rate and live embryo rate in nine groups to explore the impact of endometriosis on pregnancy outcome in vitro fertilization and embryo transplantation and Huo Xuexiaoyi Fang improving the pregnancy rate and live embryo rate.Results:1)Two weeks after establish the endometriosis mice model,there are lesions in mice subcutaneous and abdominal cavity.Campared the pregnancy rate in model group with the sham operation group and the blank group,the differences were statistically significant.(P<0.05).The live embryo rate in the three groups,the differences were statistically significant(P<0.05).2)The developed pinopodes were expressed in endometrial surface at 86.5h.The developing pinopodes were expressed at 80h,84h,85h,85.5h.The degenerating pinopodes were expressed at 87h,88h,91h,95h.Pinopodes are the important morphologic marker of endometrial receptivity,their appearance on the surface of endometrium takes place during the implantation window.3)The average stimulate ovulation number among TCM group,the model group and the sham operation group,there were significantly difference(P<0.05).The fertility rate in vitro among TCM group,the model group and the sham operation group,there were significantly difference(P<0.05).The rate of two-cell cleavage among TCM group,the model group and the sham operation group,there were significantly difference(P<0.05),there were significantly difference(P<0.05).The rate of four-cell cleavage in TCM group compare with the model group and the sham operation group,there were significantly difference(P<0.05).There were no significant differences in the blastulation rate among three groups(P>0.05).There were significant differences in the blastulation rate finally among three groups(P<0.05).4)There is one mice to fail to transplant in group 2 and group 5 respectively due to severe adhesion.The pregnancy rate of nine groups were significant differences(P<0.05).Camparing TCM-sham operation group,model-sham operation group and sham operation-sham operation group,the pregnancy rate were significantly differences(P<0.05).Camparing sham operation-TCM group,sham operation-model group and sham operation-sham operation group,the pregnancy rate were significantly differences(P<0.05).As the live embryo rate of nine groups from high to low,followed by sham operation-sham operation group,sham operation-TCM group,TCM-TCM group,TCM-sham operation group,TCM-model group,sham operation-model group,model-sham operation group,model-TCM group and model-model group.Conclusions:1)Established the endometriosis mice model with pregnancy low function successfully.2)The developed pinopodes were expressed in endometrial surface at 86.5h.3)Endometriosis can influence the average stimulate ovulation number,in vitro fertilization and embryonic development of model mice.4)Huo Xuexiaoyi Fang can improve the average stimulate ovulation number,the fertility rate in vitro and embryonic development in model mice.5)Huo Xuexiaoyi Fang can improve the pregnancy rate and live embryo rate in vitro fertilization and embryo transplantation.6)Huo Xuexiaoyi Fang can improve the receptivity of endometrium and embryo quality in the model mice. |
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