Study On The Effect And Mechanism Of DS-1-47,the Active Ingredients From A Recipe In Promoting Pregnancy

DS-1-47,the effective components of promoting embryo implantation,which extracted from a clinical promoting pregnancy recipe.The recipe is composed of Astragalus,Atractylodes,Radix Scutellariae and Radix dipsaci.DS-1-47 is a compound fraction obtained from ethyl acetate extraction and silica gel column chromatography.In this study,combined injection of PMSG and HCG to establish the embryo implantation dysfunction model to observe the effect.HE staining,Immunohistochemistry and Western blot were used to detect the expression of ANG,NF-?B,PCNA,BAX,CD31 to explore the molecular mechanism of DS-1-47 promoting pregnancy..Part 1.Effects of DS-1-47 on the embryo implantation rate and uterine morphological development of controlled ovarian hyperstimulation miceCombined injection of PMSG and HCG to establish the embryo implantation dysfunction model.After successful mating,animals were divided into model group(M)and traditional Chinese medicine group(DS-1-47)randomly.Normal mice were as normal group after mating.Mice of DS-1-47 group was orally administrated with DS-1-47 every day.Normal and model mice were orally administrated with saline.Mice were sacrificed at fifth,sixth and eighth day after mating.Pregnancy rate and the average number of implantation were calculated.The uterus of mice on fifth and sixth pregnancy day were removed and fixed with 4% paraformaldehyde for HEstaining.The results showed that DS-1-47 significantly promoted embryo implantation and pregnancy rate.It also improved uterine vascular permeability,endometrial thickness,endometrium decidualization and uterus closure.These showed that DS-1-47 had a good effect on promoting embryo implantation and improving the development of uterus.Part 2.Study on the molecular mechanism of DS-1-47 on promoting embryo implantationCombined injection of PMSG and HCG to establish the embryo implantation dysfunction model,groups and administration were done as above.The mice were sacrificed at fifth and sixth pregnancy day after mating,uterus of each mice was removed,one side was stored in a refrigerator at-70?,and the other side was fixed with 4% paraformaldehyde.Then,HE staining,immunohistochemical staining and western blot were used to detect the expression of implantation related molecules.The results showed that the expression of ANG,NF-?B,PCNA,BAX and CD31 in the endometrium of mice in DS-1-47 group during the early implantation window decreased,and the spatio-temporal expression of the above molecules from PD5 to PD6 was promoted.In addition,the expression of NF-?B in ovary on PD5 and PD6 also decreased.It shows that DS-1-47 inhibit the ANG stress pathway and the excessive proliferation and apoptosis of `endometrial cells,thus to provide a stable and suitable endometrial microenvironment for embryo implantation.