| Objective:Pre-clinical and experimental studies have demonstrated its efficacy.In order to explore its effect on early pregnancy progesterone to maintain pregnancy and further reveal its mechanism of action,the research aims at observing the influence on the uterine viscera index,the pregnancy rate,the number of implanted blastovysts,the decidua tissue morphology,the endometrial tissue morphology,the uterus tissue protease metal matrix MMP-9,the content of its inhibitor of TIMP-1 and LIF of pregnant model rats with kidney deficience and blood stasis syndrome.Preliminary this paper discusses on the mechanism of improving maternal-fetal interface immune micro-environment,which provides a theoretical basis for clinical use,provides new ideas and lays a foundation for the further development of the experiment.Methods:The adult unpregnant female SD mouse that in proestrus were randomly divided into five groups,namely blank group,model group,assisted reproduction 1 prescription by high and low dose group,wuzi group,danshen group,by vaginal smear and determination.The wuzi group and danshen group as a positive drug control group.In addition to the model group,the remaining groups lavage by hydroxyurea orally plus adrenaline hypodermic cause kidney deficiency and blood stasis composite model.At the same time modeling of kidney deficiency and blood stasis,the mouse of assisted reproduction 1 prescription by high and low dose group concentrated extract of Chinese medicine treatment of rectal administration;the mouse of wuzi group were given Wu Zi Yan Zong Wan suspension gavage;the mouse of danshen group were in the treatment of oral Danshen tablets suspension.When the kidney deficiency and blood stasis model is established,the female rats in estrus in each group were screened by the vaginal secretion,male and female rats by 2:1 ratio to be mated.Referred to as first day of pregnancy if it was found vaginal plug or vaginal smears observed sperm the second day,and then stop treating pregnant rats administered.In the 8th day of pregnant,put the rat to death after cutting the femoral artery blood,rapidly weighing the uterus tissue,fixed it in the neutral formalin fixative,observed the corresponding metrics,then calculating the index of uterine viscera,observing the pathological morphology change of the uterus endometrial tissue of the rats in each group under the microscope,and detecting the expression level of MMP-9,TIMP-1 and LIF those of values were determined by immunohistochemistry from endometrium.Results:1.The effect of assisted reproduction 1 prescription imported by rectum on the uterine viscera index of model rats with kidney deficiency and blood stasis syndrome.In model group,wet weight of uterus and organ index were significantly lower than control group(P<0.05),showed that the model of kidney deficiency and blood stasis may affect the uterine and embryonic rat model of growth and development;compared with the model group,the uterine viscera index of assisted reproduction 1 prescription high dose group,low dose group and Wuzi group were higher than those of the model group(P<0.05),suggesting that assisted reproduction 1 prescription and Wuzi Yanzong pills can promote uterus and embryo growing and development of the pregnant rat with kidney deficiency and blood stasis.There was no significant difference in the uterine wet weight and the uterine organ index of Danshen group in the model group(P>0.05).2.Effect on the pregnancy rate and blastocyst implantation number of kidney deficiency and blood pregnancy stasis rats threated by assisted reproduction 1 prescription imported by rectum.The pregnancy rate and the embryo implantation number of model group were lower than that of the blank group(P<0.05),suggest that modeling kidney deficiency and blood stasis can effect rats’ pregnancy ending,reduce the pregnancy rate and inhibit the pregnancy rats’ blastocyst growth and development.The pregnancy rate and the embryo implantation number of assisted reproduction 1 prescription high dose group were higher than that of the model group(P<0.05).The pregnancy rate and the embryo implantation number of assisted reproduction 1 prescription low dose group were higher than those in the model group,but the difference has no statistically significant(P>0.05),suggest that the assisted reproduction 1 prescription may increase the pregnancy rate and promote the development of the embryo in rats with kidney deficiency and blood stasis.3.The influence of assisted reproduction 1 prescription imprted by rectum of kidney deficiency and blood stasis model rats’ MMP-9,TIMP-1 on uterine tissueThe total MMP-9 expression area of model group rat uterus tissue was significantly lower than that of the model group(P<0.05),the sum MMP-9 expression of integral optical density of model group rat uterine tissue has higher trend than that of model group(P>0.05);compared with the model group,medicine group rat uterus tissue MMP-9 expression of the sum of the area were significantly increased(P<0.05);in addition to danshen group,other medication in each group rat uterine tissue MMP-9 expression of integral optical density of the sum compared with the model group compared to were significantly increased(P<0.05).Danshen group rats’ uterine tissue MMP-9 expression of integral optical density of the sum was higher than the model group(P>0.05).The expression of TIMP-1 in uterine tissues of the model group was significantly lower than that in the blank group(P<0.05).Except the Danshen group,in each other medication group theTIMP-1 expression of the total area compared with the model group were significantly higher than that(P<0.05).Compared with the rats in the model group and assisted reproduction 1 prescription high dose group significantly increased the expression of TIMP-1 integral optical density sum in uterine tissue(P<0.05),and in each other medication group,the rats’ uterine tissue TIMP-1 expression of integral optical density of the sum have the higher trend to model group.Blank group,assisted reproduction 1 prescription low dose group,assisted reproduction 1 prescription high dose group,Wuzi group,the area of MMP-9/TIMP-1 sum and integral optical density sum were close to 1:1.Assisted reproduction 1 prescription high dose group was the most closer one.4.The influence of assisted reproduction 1 prescription imprted by rectum of kidney deficiency and blood stasis model rats’ LIF in pregnancy uterine tissueThe expression of LIF in model group was significantly lower than that in the blank group(P<0.05).and the expression of LIF in the uterus of the blank group was higher than that in the model group(P>0.05).Compared to the model group,except assisted reproduction 1 prescription low dose group,in each other medication group,the LIF expression in the sum of the area and integral optical density sum were significantly increased(P<0.05);assisted reproduction 1 prescription low dose group rats’ uterine LIF expression has higher trend to model group(P>0.05).5.The regulation to the expression of LIF,MMP-9.TIMP-1 on kidney deficiency and blood stasis model rats’ uterine tissue by assisted reproduction 1 prescription treating.The LIF expression corresponding MMP-9.TIMP-1 expression of the total area,significant correlation at 0.01 level(bilateral);The LIF expression integral optical density of the sum and each corresponding expression of MMP-9 in the integral optical density of the sum,at the 0.05 level(bilateral)significant correlation.Conclusion:1.Treated by assisted reproduction 1 prescription through rectum before pregnancy can promote the growth of kidney deficiency and blood stasis model rats’ uterus and embryo development,promote uterine endometrial decidualization,improve the endometrial morphology,provide sufficient material basis for the embryonic development,thereby increasing the pregnancy rate and embryo implantation.2.Treated by assisted reproduction 1 prescription through rectum before pregnancy can improve the micro environment of a maternal fetal interface,increases tolerance to embryo of the maternal immune system,thus contributing to the maintenance of pregnancy,the mechanism might be:(1)Treated by assisted reproduction 1 prescription through rectum before pregnancy can improve the expression of MMP-9,TIMP-1 in the kidney deficiency and blood stasis model rats uterine tissueand,and can maintain the ratio of MMP-9/TIMP-1 in a dynamic equilibrium state,both expression level in equilibrium relative steady state,ensure the trophoblast cells of the uterine decidua moderate infiltration and moderate corresponding regulation of extracellular matrix degradation and improve the immune tolerance of the maternal fetal interface,so as to maintain the normal pregnancy.(2)Treated by assisted reproduction 1 prescription through rectum before pregnancy can improve the kidney deficiency and blood stasis model rats uterine tissue of LIF expression,on the one hand,the LIF can directly regulate cell permeability,improve the micro environment of a maternal fetal interface,and promote the growth and development of the embryo;through another increasing the content of LIF may positively modulate the expression levels of MMP-9,also influence TIMP-1 secretion and expression improve maternal fetal interface micro environment,enhance the immunotolerance at the fetal maternal interface,to facilitate the maintenance of pregnancy. |
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