| Objective:Based on the clinical efficacy and experimental study of the treatment of infertility and IVF-ET failure in the early stage of Kidney-supplementing and blood-moving Formula,in order to further explore its pregnancy-promoting effect,this experiment was conducted to observe the effect of Kidney-supplementing and blood-moving Formula on kidney deficiency and blood stasis.Endometrial histomorphology,uterine organ index,rat progesterone(P),gestational progesterone receptor(PR),lipoxin A4(lipoxin A4),LXA4),LXA4 receptor ALX,homeobox gene A10(HomeboxA10,HOXA10),insulin-like growth factor binding protein-1(IGFBP-1),preliminary study on the effect of Kidney-supplementing and Blood-moving Formula by Rectal Administration can improve the mechanism of endometrial receptivity in the implantation window of rats with renal dysplasia and blood stasis by regulating LXA4,and provide a theoretical basis for clinical drug use.Methods:Through the smear and judgment of vaginal secretions,healthy adult female nonpregnant SD rats in the early stage of estrus were randomly divided into blank group,model group,Kidney-supplementing and Blood-moving Formula low,high dose group,Chinese medicine control group and western medicine control group.Except the blank group,the rats in the other groups were given a compound model of kidney deficiency and blood stasis by intraperitoneal injection of hydroxyurea and adrenalin,and the general condition of the rats after modeling was visually observed.At the same time of modeling,except for the blank group and model group,the other groups of rats were given corresponding drugs.After the successful modeling,the rats in each group were screened out of the estrus female by vaginal secretion smear,and the cage was performed in a ratio of 2:1 for male and female rats.The second day was observed on the second day.The worms were recorded as the first day of pregnancy,and the drug was stopped after the pregnancy.The mifepristone was injected subcutaneously on the 4th day of pregnancy to form a implantation disorder model.On the fifth day of pregnancy,the rats were anesthetized with sodium pentobarbital and fixed.The abdominal aorta blood was taken by laparotomy,the bilateral uterus was quickly removed,the adipose tissue around the uterus was weighed,the uterine organ index was calculated,the uterus specimens were collected,and the morphological changes of endometrium in each group were observed under light microscope.IGFBP-1 was localized on endometrial tissue by immunohistochemical technique.The expression of LXA4 and ICFBP-1 in uterine tissue was detected by enzyme-linked immunosorbent assay(ELISA),and the expression of P in peripheral blood was quantified by real-time fluorescence.The expression of PR,ALX and HOXA10 mRNA in uterine tissue was detected by acid chain reaction.Results: 1.Effect of Kidney-supplementing and Blood-moving Formula on Endometrial Morphology and Uterine Organ Index in Rats with Kidney Deficiency and Blood Stasis Implantation.The low-and high-dose groups of Kidney-supplementing and Blood-moving Formula have a growing trend of endometrial epithelial cells.The epithelial cells are arranged neatly and evenly in the shape of long columns.The glands are rich,the glandular cavity is full of round or elliptical shape,the blood vessels are rich,and the interstitial arrangement is even and loose.The intimal morphology increased well;the uterine wet weight and uterine organ index in the low-and high-dose groups of Kidneysupplementing and Blood-moving Formula were higher than those in the model group,such as uterine wet weight and uterine organ index.2.Effect of Kidney-supplementing and Kidney-supplementing and Blood-moving Formula on the expression of P and PR in the implantation window of rats with kidneydeficiency and blood stasis.Compared with the blank group,the serum P level in the model group was significantly decreased(P<0.05),and the uterus tissue PR level was also significantly decreased(P<0.001).Compared with the model group,the serum P of the high-dose group of Kidney-supplementing and Blood-moving Formula The uterine tissue PR level was significantly increased(P<0.05).The PR level of uterus in the low-dose group of Kidney-supplementing and Blood-moving Formula was significantly increased(P<0.05),and the serum P increased(P>0.05).3.Effect of Kidney-supplementing and Blood-moving Formula on Expression of LXA4 and ALX in Uterine Tissue of Rats with Kidney Deficiency and Blood Stasis Implantation Disorder Model.Compared with the blank group,the levels of LXA4 and ALX in the uterus tissue of the model group were significantly increased(P<0.001).Compared with the model group,the expression of LXA4 in the uterus tissue of the high-dose group of Kidneysupplementing and Blood-moving Formula decreased.Significantly(P < 0.001),the expression of ALX in uterine tissue decreased(P>0.05).The LXA4 and ALX levels in the uterine tissue of the low-dose group of Kidney-supplementing and Blood-moving Formula were significantly lower(P<0.001).4.Effect of Kidney-supplementing and Blood-moving Formula on Expression of HOXA10 and IGFBP-1 in Uterine Tissue of Rats with Kidney Deficiency and Blood Stasis Implantation Disorder Model.Compared with the blank group,the HOXA10 level in the endometrial tissue of the model group was significantly decreased(P<0.001),and the expression of IGFBP-1 was also significantly decreased(P<0.05).Compared with the model group,the kidney and blood circulation were compared.The level of HOXA10 in the endometrial tissue of the high-dose group was significantly increased(P<0.001),and the expression of IGFBP-1 was also significantly increased(P<0.05).Conclusion: 1.Kidney-supplementing and Blood-moving Formula can promote the development of uterus in rats with kidney deficiency and blood stasis,increase the uterine organ index,improve the endometrial tissue morphology in the planting window,and improve the endometrial receptivity of rats.2.The mechanism of assisting pregnancy by Kidney-supplementing and Blood-moving Formula may be: by up-regulating the level of serum P and uterine tissue PR in the planting window,down-regulating the expression of LXA4 and ALX in uterine tissue,and promoting the increase of HOXA10 and IGFBP-1 in endometrial tissue,thereby improving the large Rat endometrial receptivity,ready for embryo implantation. |
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