The Expression And Significance Of LPAR3and HB-EGF In Endometrial Implantation Window Period Of RIF Patients

BackgroundWhen the embryo is on the state of implantation, the endometrium is also switched to the state of accepting an embryo, this short period which allow embryo implantation named implantation window. Over the years, the number and quality of embryos have been optimized gradually by improving the technology of controlled ovarian hyperstimulation and laboratory, Especially the application of intracytoplasmic sperm injection (ICSI) technique which solves a lot of problems in embryo quality caused by sperm and egg union in1992.But the implantation rate of IVF-ET is still around30%, an important reason is endometrial receptivity defects in some infertile women when implantation window, affecting the successful implantation of the embryo. It is believed that the transplant number≥3or transplant good quality embryos≥10are unable to pregnancy could be called repeated implantation failure by most scholars at present. Usually, no clear reason of infertility could be found in such patients, it is a hot and difficult issue that how to predict and improve RIF patients’ endometrial receptivity in order to enhance the pregnancy rate of next assisted reproduction. For nearly thirty years, domestic and foreign scholars have done a lot of research in endometrial receptivity, and found a series of hormones, cell factors and growth factors which can influence endometrial receptivity. These factors constitute a large, complicated and orderly molecular network, two-way regulating and interact with each other. We choose LPAR3and HB-EGF as starting points to do research in endometrial receptivity this time.ObjectiveDiscussing the possible mechanism of descend endometrial receptivity in RIF patients, and searching for suitable evaluation index to provide the theory basis on therapeutic measures which can improve the endometrial receptivity.Materials and methods23RIF patients are chosen as experimental group, and31patients who pregnancy after the first IVF-ET treatment as control group. They are all come from reproductive medicine center of Zhengzhou university third affiliated hospital from October2010to August2011.Collection of Endometrium:Monitoring follicular by transvaginal color doppler sonography in the menstrual cycle before ovulation induction, then doing pre-implantation seven days after ovulation. Endometrium of implantation window is collected in the meanwhile. The endometrium specimens divide into two, the first one is fixed in10%neutral buffered formalin, dehydrated by gradient ethanol, embedded by paraffin, then make into slices for pathological examination (confirm histology installment and eliminate endometrial lesions) and determining LAR3and HB-EGF by immunohistochemical SP method. Freeze the fresh second one in liquid nitrogen for determining LAR3and HB-EGF by semiquantitative RT-PCR.Collection of peripheral blood:Cubitsvenous blood is extracted on an empty stomach in the morning when endometrium is collected, then examine the E2and P value by chemical luminescence method. It is recorded detailedly in patient age, the number of transplant cycle, infertility cause and type, basis status of reproductive endocrine, E2and P value in implantation window, endometrial thickness, controlled ovarian hyperstimulation situation in IVF cycle, the number of retrieved oocytes and embryo quality.SPSS17.0statistical analysis software was used for date analysis.The quantitative data which meet normal distributionbetween two groups was analysised by t test sample, others by rank sum test. The qualitative data was analysised by Chi-square test. Correlation analysis was used Pearson. Bilateral a=0.05for inspection standards.Result1. There are on significant differences in BMI, infertility cause and type, basis levels (the second day to the forth day in menstrual cycle) of FSH, LH and E2, the level of E2in implantation window, pituitary down-regulation scheme, embryo quality in laboratory. But the patients’ age, days and doses of Gn are higher than control group significantly(31.652±3.284vs.29.839±2.423,P=0.023)(11.261±1.137vs.10.484±1.262, P=0.024)(2311.967±565.290vs.1930.694±511.993,P=0.012). And the level of P in serum is lower than control group significantly (11.365±3.612vs.15.039±4.127,P=0.001)2. Both LPAR3and HB-EGF in patients’ endometrial of two groups are positive expression; the expression of them is consistency both in parts and characteristics. They are typical top plasma secretion and distributed mainly in endometrial glands epithelial cells and stromal cell. The average optical density values of LPAR3and HB-EGF in RIF group was are lower than control group significantly (LPAR3:0.255±0.105vs.0.359±0.102,P=0.001; HB-EGF:0.298±0.126vs.0.55410.197,P<0.001).3. The loss expression of LPAR3and HB-EGF mRNA in RIF group are significantly than control group (LPAR3:0.481±0.167vs.0.647±0.177,P=0.001; HB-EGF:0.961±0.312vs.1.514±0.346.P<0.001). 4. The expression of LPAR3and HB-EGF mRNA in these two groups are negative correlation to patients’age (LPAR3:r=-3.575,P<0.001; HB-EGF:r=-6.025,P<0.001). According to current data, we still do not think there are significantly correlation in expression of LPAR3and HB-EGF mRNA to Gn days, Gn doses, retrieved oocytes number, E2and P value in implantation window.Conclusions1. The expression of LPAR3, both in parts and characteristics, is conformity with HB-EGF, so it could be a good reference index to predict endometrium receptivity.2. Lower P value, the loss expressions of LPAR3, HB-EGF in endometrium may be the reason which caused repeated implantation failure in infertility.3. The expressions of LPAR3and HB-EGF decreased with the increase of age.