The Effect Of High Progesterone On Endometrial Receptivity By Regulating The Expression Of TGβ3,EMX2 And PCAF Through HOXA10

As mankind enters the 21 st century,infertility has become the third largest disease worldwide.In the treatment of infertility,assisted reproductive technology(ART)is an important method of infertility treatment,and in vitro fertilization embryo transfer technology(IVF-ET)is an important part of ART.Embryo implantation is a prerequisite for a successful pregnancy,and a well-developed embryo and a matching endometrium in a receptive state are essential for embryo implantation and continued development.However,in recent years,studies have found that in the application of IVF-ET,even when the blastocyst is in good condition,nearly one-third of the patients still fail to implant successfully,suggesting that the endometrial receptivity plays a key role in embryo implantation.The expression of HOXA10 in the endometrium is positively regulated by progesterone,which in turn regulates the functions of related genes ITGβ3,EMX2 and PCAF to maintain endometrial receptivity.In assisted reproductive technology,due to the application of controlled ovulation stimulation drugs,multiple follicles mature at the same time,and produce estrogen and progesterone that exceeds the normal human physiological range,which may affect the outcome of pregnancy.The previous research of our group found that the high progesterone environment reduced the expression of HOXA10,suggesting that the expression of HOXA10 gene may be inhibited by high progesterone,which in turn affects the endometrial receptivity and pregnancy outcome.Based on this,it is necessary to further explore the specific regulatory mechanism of HCG daily high progesterone level on HOXA10,ITGβ3,EMX2 and PCAF in the IVF-ET cycle,so as to provide certain theoretical basis for the effective treatment strategy of clinical hyperpregnant patients.This subject conducts research from two levels of clinical sample experiment and cell experiment:(1)Clinical sample research,choose to go to the Department of Reproductive Medicine of the First People’s Hospital of Yunnan Province to receive IVF or ICSI due to male factors or female fallopian tube factors and cancel fresh transplants Patients,according to HCG day serum estrogen stimulus level,set the sampled book into three groups: high progesterone group(P≧2.5 ng/ml)(30 cases);high estrogen group(E2≥5210 pg/ml)(30 cases));High estrogen combined with high progesterone group(E2≥5210 pg/ml,P≥2.5 ng/ml)(30 cases).In addition,patients with spontaneous ovulation were collected as a control group(P≦1.05 ng/ml)(30cases).Endometrial tissue was collected within 7-8 days after ovulation by controlled ovulation induction or B-ultrasound detection,and the endometrial tissue was collected by immunohistochemistry(IHC),protein Western blot(WB)and real-time fluorescent quantitative PCR(q RT-PCR)were used to detect the expression of HOXA10,ITGβ3,EMX2 and PCAF genes.The results showed that compared with the control group,the expressions of HOXA10 and ITGβ3 protein in the high progesterone group and the high estrogen combined with high pregnancy group were significantly reduced(P≤0.05),and the EMX2 and PCAF proteins were significantly increased(P≤0.05),while the high estrogen group HOXA10 The expression of,ITGβ3,EMX2 and PCAF did not change significantly.It is suggested that high progesterone inhibits the expression of HOXA10 and ITGβ3,and promotes the expression of EMX2 and PCAF;(2)Cell experiment research,Ishikawa cells are compared with 10-5,10-6,10-7,10-8 and 0 M/L progesterone,respectively After co-cultivation for 24 hours,Western Blot and RT-PCR were performed to detect the relationship between the expression of HOXA10,ITGβ3,EMX2 and PCAF genes and progesterone.At the same time,over expression of HOXA10 was used to detect the expression of PCAF,ITGβ3,PCAF and EMX2.Results: When the progesterone level is less than 10-7,the expression of HOXA10 and ITGβ3 increases significantly with the increase of the progesterone level,and the expression of PCAF and EMX2decreases;when the progesterone level is greater than 10-7,the expression of HOXA10 and ITGβ3 decreases significantly with the increase of the progesterone level,PCAF and EMX2 expression increased.And after over expression of HOXA10 gene for 48 h,the expression of ITGβ3 was significantly increased,and EMX2 and PCAF were significantly decreased.It shows that high progesterone levels can inhibit the expression of HOXA10 and its target gene ITGβ3,and promote the expression of EMX2 and PCAF.Summarizing and analyzing the experimental results,it is concluded that the high progesterone environment may reduce the expression of HOXA10,decrease the expression of ITGβ3,and increase the expression of PCAF and EMX2,which is unfavorable to the endometrial receptivity,thereby affecting the outcome of IVF pregnancy.