| Background and ObjectiveThe birth of modern assisted reproductive technology (ART) has brought new methods for the clinical treatments of infertility, and helped countless families to solve the trouble without children.The first test tube baby came to this world in 1978, by in vitro fertilization and embryo transfer (IVF-ET) in natural ovulation cycle. Then, fertility experts obtained more eggs by super ovulation, thus increasing the clinical pregnancy rate of IVF-ET. With the rapid development of ART, they studied out the controlled ovarian hyperstimulation (COH), which significantly increased the number of eggs. In assisted reproductive technology, they also use hormone replacement therapy (HRT). HRT adjusts the endometrium to the appropriate state before the frozen-thawed embryo transfer (FET), which applies to patients with irregular menstrual cycles.ART has been available for more than thirty years, although the in vitro fertilization rate can be as high as 90%, the clinical pregnancy rate is still low, about 30%-60%. So, many experts questioned the above mentioned treatments can make the patient's sex hormones exceed physiological levels, affecting the body's micro-environment. The endometrial receptivity decreases, resulting in failure of pregnancy. Therefore, in recent years, the study of endometrial receptivity has been paid more and more attention to in the field of assisted reproduction.In a menstrual cycle, the embryo can be implanted into the endometrium in a very short period, which is termed "implantation window". Endometrial receptivity refers to the ability of endometrium to accept embryo. Embryo implantation is accomplished by the interaction of multiple factors, such as leukemia inhibitory factor (LIF), interleukin-1 beta (IL-1?), integrin alpha v beta 3(integrin av?3), etc. A number of studies have indicated that, the period of high expression of LIF, IL-1? and integrin av?3 is the same as implantation window. Therefore, these factors can be considered as markers of endometrial receptivity. Previous animal experiments found that hormone replacement therapy, controlled ovarian hyperstimulation, and super ovulation decreased mouse endometrial receptivity. Their degrees increased gradually.The objective of this study was to observe the expression of the above factors in human endometrium during the implantation window, based on different treatments (super ovulation, controlled ovarian hyperstimulation and hormone replacement therapy). Then we can explore these commonly used clinical treatments'effects on endometrial receptivity, thus providing a more reliable theoretical basis for the optimization of clinical treatments and improving the clinical pregnancy rate.Materials and methods1. Experimental materialsExperimental groups were patients who were under treatments in IVF/ICSI cycles or HRT-FET cycles, but canceled the implantation for some reasons in our center during June 2014 and December 2015. Control groups were women who were in natural ovulation cycles without any drug. We collected their endometriums by disposable sterile endometrial collecting tubes, according to the following grouping methods.Super ovulation group:in super ovulation cycles,5 and 6 days after oocyte retrieval were super ovulation 5d group and super ovulation 6d group respectively (5 cases each).COH group:in controlled ovarian hyperstimulation (COH) cycles,5 and 6 days after oocyte retrieval were COH 5d group and COH 6d group respectively (10 cases each).HRT group:in hormone replacement therapy (HRT) cycles,5 and 6 days after injecting progesterone were HRT 5d group and HRT 6d group respectively (5 cases each).Control group:in natural ovulation cycles,5 and 6 days after ovulation were natural ovulation cycle 5d group and natural ovulation cycle 6d group respectively (10 cases each).Each patient was sampled only one time instead of two consecutive days. In each case, the endometrium was divided into two parts, one for immunohistochemistry, and the other was sent to the pathology department.2. Experimental methodsImmunohistochemical SP method was used to detect the localizations and semi quantitations (IOD values) of LIF, IL-1?, integrin ?v?3 in human endometrium.3. Statistical methodsAnalysis by SPSS 22.0 software, measurement data were expressed as x±s, tested by independent samples T-test or multiple samples One-Way ANOVA (pairwise comparison using the Bonferroni method), P<0.05 was statistically significant.Results1. General information of patients in each group (mean age, duration of infertility, BMI, basal FSH and thickness of the sample) had no significant difference.2. The pathological results of the endometrium in each group showed that, the secretory phase endometrium, there was no special pathological condition.3. LIF, IL-1? and integrin avP3 were mainly expressed in the cytoplasm of luminal and glandular epitheliums. These three proteins' expression in the endometrium were weakly positive in super ovulation group and COH group, positive in HRT group, and strong positive in natural ovulation cycle group.4. In natural ovulation cycle 5d group, HRT 5d group, COH 5d group and super ovulation 5d group, the IOD values of LIF were 149.95±14.51,133.82±6.61, 117.51±10.87,92.43±12.71 respectively. The IOD values of IL-1? were 143.97±16.36,124.56±8.58,104.39±12.24,79.33±18.69 respectively. The IOD values of integrin av?3 were 141.61±9.69,126.97±8.91,112.92±9.92,78.45±18.69 respectively.The expression of these three proteins in the 5d groups was natural ovulation cycle 5d group> HRT 5d group> COH 5d group> super ovulation 5d group (P<0.01).5. In natural ovulation cycle 6d group, HRT 6d group, COH 6d group and super ovulation 6d group, the IOD values of LIF were 157.01±10.35,139.17±5.79, 111.71±17.02,81.35±9.39 respectively. The IOD values of IL-1? were 148.83±10.72, 130.74±7.01,94.85±20.53,66.79±10.79 respectively. The IOD values of integrin av?3 were 149.36±11.48,131.91±9.99,105.82±18.41,70.12±6.44 respectively.The expression of these three proteins in the 6d groups was natural ovulation cycle 6d group> HRT 6d group> COH 6d group> super ovulation 6d group (P<0.01).6. The expression of these three proteins had no significant differences between super ovulation 5d and 6d groups, as well as between COH 5d and 6d groups, between HRT 5d and 6d groups, and between natural ovulation cycle 5d and 6d groups(P> 0.05).Conclusions1. LIF, IL-1? and integrin avP3 can be markers of human endometrial receptivity during the implantation window.2. All the super ovulation, COH and HRT treatments can do harm to the endometrial receptivity, but the weakening degree is different. HRT is better than COH and super ovulation, and COH is better than super ovulation.3. In the cycles of ovarian stimulation, we should fully assess the endometrial receptivity. If it decreases, we should freeze all embryos. Then we choose FET in natural ovulation cycle or HRT cycle. |
PDF Full Text Request