The Expression And Study On Pinopode And Intergrin β3 And Osteopontin In Endometrial Implantation Window Of Women With Polycystic Ovary Syndrome

Polycystic ovary syndrome (PCOS) is one of the common endocrine disorders of women during their reproductive age and it is the major cause of anovulatory infertility. The endometrium of patients with PCOS is insensitive to estrogen or progestin, which may affect blastocyst implantation. Although few patients of PCOS have their own ovulation, the rate of pregnancy is much lower than common women. High ovulation rate but low pregnancy rate will be found in PCOS during controlled ovarian hyperstimulation, of which that reduced response of endometrium to blastocyst may be in charge. Some of them have to turn to IVF or IVM when failing to conceive a child after all kinds of therapies for infertility. Cryopreservation of embryos for future transfer in an artificial cycle has been an effective alternative for patients with PCOS when they have excess embryos during IVF/IVM or they are in high risk of OHSS during IVF. How to apply appropriate estrogen and progestin to prepare endometrium for better receptivity remains elusive. The endometrial receptivity is one of the most factors of conceiving a baby. Recent researches have demonstrated that the pinopodes can be the best morphological marker to evaluate the endometrial receptivity, and the presence of some endometrial proteins can better show the functional status. The patterns of intergrin (especiallyαvβ3) and osteopontin expression in human endometrium exhibit cycle-dependent expression with temporal characteristics. They appears in endometrium postovulatory day 6-9, corresponding to the time of embryo attachment, and has been generally accepted as a marker of endometrial receptivity. Reducedαvβ3 and osteopontin expression during the implantation window have been reported in connection with luteal phase deficiency (LPD), minimal or mild endometriosis, communicating hydrosalpinx and unexplained infertility. Therefore, choosing the endometrium of patients with PCOS as objects, and from two parts of ways (morphology and immunological related factors) to understand the status of endometrial receptivity in order to elucidate the cause of the low pregnancy rate and seek the effective resolvent.There are two parts in the study of the endometrial receptivity in the women with PCOS during the implanted window.Part 1 The Expression and Study on Pinopode in Endometrial Implantation Window of Women with Polycystic Ovary Syndrome ObjectiveTo observe the ultramicrostructure of endometrial pinopodes of PCOS patients during the implantation window and the effects of the development.MethodsThe study was carried out between 15 typical women with PCOS and 15 normal controls using a case-control study. Continuous observation of the development of follicles from the M10, and endometrial biopsies and vein blood were achieved on the day of LH7. Human endometrium was achieved by endometrial taker, and routine HE staining was accomplished in all women. Morphology was detected by light microscope and all samples were examined under SEM. Serum E2 and P were measured by chemiluminoimmunoassay (CLIA). Endometrial thickness and pattern was assessed in trans-vaginal ultersonography on the ovulating day. Therefore, the objective is to elucidate the association among the development of pinopodes and E2,P and E2/P and endometrial thickness and pattern. Compare with normal fertility women, seeking the best indicators of morphology and ultersonography to evaluate the endometrial receptivity of PCOS.Results1 The clinical characteristics of cases15 cases of each group, and there was no significant difference between the two groups with respect to age and body mass index(BMI) (P>0.05). There were significant differences of the LH/FSH and T between the experiment and control group (P<0.05).2 Routine HE staining of endometriumThe glands were cured and abundant of control group; the glandular cavity was larger than the experiment group. There was much acidophilia secretion from the glandular epithelium. Sometimes we could see the helicine arteries and veins in the endometrium. Interstitial was edemaed. There were little glandular cells and less secretion. Some inflammatory cells infiltrated into the glandular cavity, and for some glandular epithelium, there was the tendency of autolysis and apoptosis. All of 15 samples of control group were in-phase. 10(66.67%) samples of experiment group were in-phase, and there were 4 in early secretory phase and 1 in proliferative phase (33.33%). Compared with the control group, there was significant difference (P<0.05).3 The pinopodes of scanning electron microscopyOf the control group, the cells of endometrium developed prosperously and ranked orderly. The pinopodes were largely fully developed, which was well-stacked and smooth with well defined borders resembling a mushroom distributed in the endometrial surface. Under low power lens, the cells ranked disorderly, the ciliated cells and microvillus had lost the normal proportion. Some cilia were absent, or lodging, or fine, or short and small, or gathered together disorderly, and lost the normal functions. Under the high power lens, the microvillus was very fade and small, or flat. Most cases of experiment group had the pinopodes without 1 sample. All samples of control group had been detected the pinopodes, for the individual reason, 10 cases were fully developed pinopodes, and 3 were developing piopodes, and the remaining 2 were regressing pinopodes. The abundance of pinopodes of the control group was significantly higher than that of the experiment group (P<0.01).4 Thickness of endometrium and pattern of the ovulating dayThere was no significant difference between the two groups with respect to the thickness of endometrium (10.53±2.01vs10.57±2.19) mm. The endometrial patterns of experiment group were: 4 of A; 7 of B and 4 of C; for the control group, they were: 10of A; 5 of B and 0 of C. There was significant difference between the two groups with respect to the pattern of endometrium (P<0.05).5 Serum E2 level(pg/ml), P level (pg/ml), and E2/P of the implantion windowSerum E2 level of experiment group was significantly higher than that of control group (P<0.01); while serum P level of experiment group was significantly lower than that of control group (P<0.01); Serum E2 /P ratio of experiment group was significantly higher than that of control group (P<0.05).Conclusions1 Pinopodes of PCOS patients during the implantation window do not fully develop, and the abundance of pinopodes decreased, and the endometrial receptivity was decresed.2 The thickness of endometrium of PCOS patients was as the normal healthy women but the endometrial pattern, which might be one of the reasons inducing the low pregnancy.3 The disorder of E2/P valve played an important role in the decrecion of pinopodes in endometium of PCOSpatients, which in harmful to implantation of embro.Part 2 The Expression and Study on Intergrinβ3 and Osteopontin in Endometrial Implantation Window of Women with Polycystic Ovary SyndromeObjectiveTo investigate the expression ofανβ3 and OPN of endometrium of PCOS and access the correlation betweenανβ3 and OPN.MethodsThe samples were achieved like above.ανβ3 and OPN were measured by SP immmunohistochemical techniques. Immunostaining outcome was analyzed by the Motic Med 6.0 medical image analysis system.Results1 Expression ofανβ3 and OPN in endometriumImmumostaining of Intergrinανβ3 was located in cytoplasm of the endometrial epithelial cells and glandular epithelial cells. In the control group, Intergrinανβ3 was strongly expressed, and weak immunostaining was observed in experiment group. There was significantly difference between the two groups (P<0.01). Immumostaining of osteopontin was located in cytoplasm of the endometrial epithelial cells, glandular epithelial cells, and some interstitial cells, it was positively expressed. The osteopontin of the control group was strongly expressed, while weakly expressed in experiment group. There was significant difference between the two groups (P<0.01).2 The correlation analysis of Intergrinανβ3 and OPN Immumostaining of Intergrinανβ3 and OPN had strong positive correlation in control group(P=0.0007, r=0.7746);and the same positive correlation in experiment group(P=0.0333, r=0.5509).Conclusions 1 Immumostaining of Intergrinανβ3 is located in membrane andcytoplasm of the endometrial epithelial cells and glandular epithelial cells, while osteopontin is located in cytoplasm of the endometrial epithelial cells, glandular epithelial cells, and some interstitial cells, and it was positively expressed.2 The expressions of endometrial Intergrinανβ3 and osteopontin are significantly lower than normal healthy women, which demonstrate that the low endometrial receptivity of PCOS patients.3 Positive correlations between Intergrinανβ3 and osteopontin makes possible that osteopontin be a good biomarker of endometrial receptivity.