| [Objective]Based on the pre clinical research and experimental research of assisted pregnancy I prescription,the experimental research on assisted pregnancy I prescription to improve pregnancy of kidney deficiency and blood stasis rats before pregnancy by rectal administration.Observe assisted pregnancy I prescription pre pregnancy by rectal administration in improving early pregnant rats' corpus luteum,reproductive hormone and its receptor,protein molecules and endometrial decidua-lization,to explore its possible mechanism of action for it rectal administration of kidney deficiency and blood stasis infertility patients,around IVF-ET patients with clinical application to provide data and theoretical support.[experimental method]Sixty female SD rats with proestrus mature unmated were selected by vaginal smear,randomly divided into blank group,model group,assisted pregnancy I prescription low dose group,assisted pregnancy I prescription high dose group,five sub group,Danshen group.model group and the treatment groups were established deficiency of kidney and blood stasis model by hydroxyurea and epinephrine.In modeling at the same time,the treatment group received intervention,after successful modeling,female rats in estrus caged found vaginal suppository and vaginal smear found sperm recorded on day 1 of pregnancy,stop drug and cage rearing,and were recorded as the first day of pregnancy when found vaginal or sperm by vaginal smear,stop drug and cage rearing,femoral artery blood sampling on pregnancy eighth days,detecting serum E2 and P levels by radioimmunoassay.the rats were.killed by cervical dislocation after then,obtained from rat uterus,egg nest,counting the rats pregnancy number and implantation points,observated of morphological characteristics of each rat corpus luteum and endornetrial decidual tissue under the microscope and detected the expression of endometrial decidua in ER,PR,LN and integrin(33 by immunohistochemical method after staining.[result]1.the effect of assisted pregnancy I prescription on pregnancy rate,implantation points of kidney deficiency and bloodCompared with the blank group,pregnancy rate and the number of blastocyst implantation of the model group ratssignificantly reduced(P<0.05);compared with the model group,pregnancy rate and the number of blastocyst implantation of assisted pregnancy I prescription(14.6g/kg)group rats increased significantly(P<0.05).2.Effect of assisted pregnancy I prescription on the corpus phase on rats with kidney deficiency and blood stasis at the eighth days of gestationCompared with the blank group,corpus luteum total area and r/R of model group rats significantly reduced(P<0.01-0.05);Compared with the model group,corpus luteum total area of assisted pregnancy I prescription(7.3g/kg)group rats,corpus luteum total area and r/R of assisted pregnancy I prescription(14.6g/kg)group rats increased significantly(P<0.01-0.05);Compared with five sub groups,corpus luteum total area of assisted pregnancy I prescription(14.6g/kg)group rats increased significantly(P<0.05);compared with Salvia miltiorrhiza group,corpus luteum total area of assisted pregnancy I prescription(7.3 g/kg)group and assisted pregnancy I prescription(14.6g/kg)group rats increased significantly(P<0.05).3.Effect of assisted pregnancy I prescription on the endometrium in eighth days of pregnancy in rats with kidney deficiency and blood stasisThe blank group rats' decidual tissue by endometrial stromal cells were polygonal or round like epithelial cells,cells arranged tightly embedded,transparent,the nucleus was oval,located in the central,blue staining,nucleolus is big and clear,non nuclear apoptosis,abundant cytoplasm staining,abundant blood vessels.Compared with the blank group,uterine decidua most of the decidual tissue of rats in model group irregular cell morphology,cytoplasmic edema,cells arranged loosely,part of the core part of the nucleus disappeared,apoptosis,the number of blood vessels was significantly reduced(P<0.01);compared with the model group,decidua cells of assisted pregnancy I prescription(7.3g/kg)group rand assisted pregnancy I prescription(14.6g/kg)group rats were polygonal or round like epithelial cells arranged tightly,transparent,the nucleus was oval,located in the central,blue staining,nucleolus is big and clear,non nuclear apoptosis,cytoplasm rich,light staining,decidua vessels increased(P<0.01);Compared with the model group,five sub group and Danshen group rats uterine decidua cell morphology is regular,cytoplasmic swelling,arranged in a dense and a small number of nuclei disappear,abundant cytoplasm,pale staining,blood vessels than the rich(P>0.05).the increase of the blood vessel of the uterine vessels in the eighth day of assisted pregnancy I prescription(14.6g/kg)group rats was better than that of the five sub group(P<0.05).4.the effect of assisted pregnancy I prescription on rats' serum E2,P levels with kidney deficiency and blood stasis syndrome on the eighth days of pregnancyCompared with the blank group,erum E2,P levels of model group rats were significantly lower(P<0.05-0.01);compared with the model group,serum E2,P levels of assisted pregnancy I prescription(14.6g/kg)group rats and serum P levels of five sub group rats were significantly increased(P<0.05-0.01).5.the effect of assisted pregnancy I prescription on ER and PR of rats with kidney deficiency and blood stasis on the eighth day of pregnancyCompared with the blank group,the model group rats' uterine decidua ER,PR total area and integral optical density o were decreased significantly(P<0.01),Compared with model group,uterine decidua ER,PR' area sum and integral optical density of assisted pregnancy I prescription(7.3g/kg)group and assisted pregnancy I prescription(14.6g/kg)group rats increased significantly(P<0.05-0.01),uterine decidua PR' integral optical density of danshen group rats increased significantly(P<0.05);Compared with five sub groups,uterine decidua PR integral optical density of assisted pregnancy I prescription(7.3g/kg)group rats increased significantly(P<0.01);compared with danshen group,the expression of uterine decidua ER.PR of assisted pregnancy I prescription group rats were significantly increased(P<0.05-0.01).6.the effect of assisted pregnancy I prescriptionon LN on integrin ? 3 on the uterus of the rate of kidney deficiency and blood stasis on 8th day pregnancycompared with the blank group,uterine decidua LN area sum and integral optical density of the model group rats were decreased significantly(P<0.01);compared with the model group,uterine decidua LN area sum and integral optical density of assisted pregnancy I prescriptionon(7.3g/kg)group and assisted pregnancy I prescriptionon(14.6g/kg)group rats were increased significantly,and uterine decidua ntegrin?3 area sum of assisted pregnancy I prescriptionon(14.6g/kg)group rats were increased significantly(P<0.05-0.01),uterine decidua LN total area of five sub group rats were increased significantly(P<0.05).[Conclusion]Assisted pregnancy I prescriptionon(tonifying kidney and blood activating therapy)pre pregnancy by rectal administration can improve endometrial decidualization and angiogenesis of kidney deficiency and blood stasis model ratsand increase the number of blastocyst implantation and improve the pregnancy rate,its possible mechanism were as follows:1.Assisted pregnancy I prescriptionon can promote corpus luteum of kidney deficiency and blood stasis model rats development,increase the area of the corpus luteum,curative effect is better than that of single tonifying kidney medicine with the function of promoting blood circulation method of traditional Chinese medicine,improve luteal shape,so as to promote the secretion of E2,P.2.Assisted pregnancy I prescriptionon can improve the expression of uterine decidua ER,PR,LN,integrin?3 of deficiency of kidney and blood stasis model rats,ER,PR were mediated by E2 and P of physiological effects.LN were combined with integrin?3,integrin and nonintegrin family proteins,which adhesive cell into the matrix and maintain the decidua of basement membrane structure and function,the trophoblast continuously to the decidua in invasion,to play the normal adhesion function,inhibiting blastocyst excessive infiltration,so as to make normal embryo implantation... |
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