Influence Of Increasing Endometrium Zhu Yun Recipe On Endometrial Receptivity Of The COH Mice In The Implantation Window

Objective:To observe the influence of Increasing Endometrium Zhu Yun Recipe on tissue morphology、sex hormone and its receptor and local factors of the COH mice endometrium in the implantation window and the number of implantation, analyze its effects, and to explore its mechanism in molecular and gene level, and to provide experimental basis for Increasing Endometrium Zhu Yun Recipe assisting IVF-ET to increase pregnancy rate.Methods:SPF clean healthy female Kunming non-pregnant mices were the research objects in the study, through the establishment of controlled ovarian hyperstimulation mouse model, Increasing Endometrium Zhu Yun Recipe was used in experimental study.Healthy Kunming female non-pregnant100mices, a week after breeding, were randomly divided into5groups, with20rats in each group, namely blank group (physiological saline group)、 model group (GnRHa+HMG+HCG group、GnRHa long period inducing ovulation scheme)、positive control group (GnRHa+HMG+HCG+aspirin group)、 low dose group (GnRHa+HMG+HCC+Increasing Endometrium Zhu Yun Recipe low dose group) and high dose group (GnRHa+HMG+HCG+Increasing Endometrium Zhu Yun Recipe high dose group). Model group、positive control group、low dose group and high dose group of mices were established the COH mice model:in the third day of estrous cycle, the timing intraperitoneal injection of Alarelin40μg/100g weight was at nine every morning, continuous nine days, on the ninth day injecting HMG0.01IU/g weight was used to induce ovulation at the same time, and after48h injecting HCG0.1IU/g body weight was used to the further maturation of oocytes.1When mices of model group, positive control group, low dose group and high dose group in Chinese medicine were injected of HCG and the blank group rats in estrus,10mices in each group were randomly taken out, according to female:male=1:1proportion, were respectively closed cage. On the next morning9:00vaginal suppository was checked, vaginal plug for the first day of pregnancy, and so on. Mices were killed at9a.m. on the eighth day after closing cage. The uterus was opened out and the both sides of the uterus were opened and the number of implantation was counted. 2The rest10mices of model group, positive control group, low dose group and high dose group after injection of HCG, in48h, blank group after ovulation (vaginal smear showed signs of ovulation)48h, were taken off the neck to death in the implantation window and materials were collected and indexes were determined. Morphological changes of endometrium were observed by HE staining, and the content of endometrial E2and P were detected by ELISA, and the expressions of endometrial ER、PR、integrinβ3、vascular endothelial growth factor (VEGF) and homeobox factor (HOXA10) were detected by immunohist-ochemistry. Leukemia expression level was detected by reverse transcrip-tion polymerase chain reaction.Results:1Compared with the blank group, in the model group, endometrial glandular dysplasia was small and straight, and the number of glands decreased, and glandular cavity was mild expanding and bendind. Glands and stroma growth was not synchronous. The expressions of endometrial E2、P、E2/P、ER、PR、integrin β3、 VEGF. HOXA10and LIFmRNA significantly reduced (P<0.01), but the number of implantation had no statistical significance (P>0.05).2Compared with the model group, in the positive control group, the endometrial morphological changes slightly improved, and the expr- essions of endometrial E2、P、ER、PR、integrinβ3、 VEGF and HOXA10were significantly different (P<0.01), and the number of implantation significantly increased (P<0.01), but the imbalance of E2/P was not corrected (P>0.05) and LIFmRNA expression was of no significant difference (P>0.05). Changes of endometrial morphology had been significantly improved in the low dose group, and the expressions of E2、P、E2/P、ER、PR、leukemia inhibitory factor LIFmRNA、integrinβ3、 VEGF and HOXA10in the endometrium were significantly different (P<0.01), and the number of implantation significantly increased (P<0.01). Changes of endometrial morphology in high dose group had been significantly improved, the expressions of endometrial E2、P、E2/P、ER、 PR、integrinβ3、VEGF、HOXA10and LIFmRNA had significant difference (P<0.01), and the number of implantation significantly incre-ased (P<0.01).3Compared with the positive control group, in the low dose group changes of endometrial morphology had been significantly improved, similar to the blank group, and the number of endometrial gland morphogenesis of rules increased, and glandular cavity expansion bend significantly. Glands and stroma synchronously developed. There were significant difference in the expressions of endometrial E2/P、ER、PR、 LIFmRNA、integrinβ3、VEGF and HOXA10(P<0.01), and there was difference in the expression of E2(P<0.05), and the number of implantation significantly increased (P<0.01); the expression of end-othelial P was of no difference (P>0.05). Morphological changes of endometrial tissue in high dose group were significantly improved, and the expressions of endothelial E2、P、E2/P、ER、PR、integrinβ3、VEGF、 HOXA10and LIFmRNA had significant difference (P<0.01), and the number of implantation significantly increased (P<0.01).4Compared with the low dose group, in high dose group there was no significant difference in morphological changes of endometrial tissue, similar to the blank group; and the expressions of endometrial E2、P、PR、 integrinβ3and HOXA10had significant difference (P<0.01), and the expressions of VEGF had difference (P<0.05), and the number of implantation significantly increased (P<0.05), but in the expressions of E2/P and ER and LIFmRNA there were no significant differences (P>0.05).Conclusions:1Increasing Endometrium Zhu Yun Recipe can effectively prevent or correct adverse effects on the mice endometrial morphology from controlled superovulation.2Increasing Endometrium Zhu Yun Recipe can significantly improve the low levels of E2and P caused by inadequate luteal function from GnRHa down regulation, improve the balance of E2/P, and increase the expressions of the COH mice endometria ER and PR.3Increasing Endometrium Zhu Yun Recipe can significantly increase the expression level of the COH mice endometrial LIFmRNA, integrinβ3, VEGF and HOXA10.4Increasing Endometrium Zhu Yun Recipe can significantly impr-ove the endometrial receptivity of COH mice and increase the number of implantation, the effect is better than that of aspirin, safe and reliable.