Study On Expression Of Nucleolar Channel System, GPR30, ERα, ERβ And PR In Endometrium During Window Of Implantation Of Women With Polycystic Ovary Syndrome

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorder of women during their reproductive age, which is a major cause of anovulatory infertility. Many PCOS patients are combined with infertility, mostly because of ovulation failure. But some other PCOS patients with normal ovulation who still got low pregnancy rate, which maybe due to their lower Endometrial Receptivity. At last, most infertile patients with PCOS have to turn to IVF ( In vitro fertilization-embryo transfer)technology in order to have their healthy baby. At this time, Endometrial Receptivity directly determines whether a satisfactory embryo is able to successfully implant in human uterus, which have already drown many researchers'attention. Influence of Estrogen Receptora(ER) and Progesterone Receptor(PR) to endometrium in implantation period is always the hotspot of these researches. Recently, Nucleolar Channel System(NCS) has caused more and more interest as a morphology marker to evaluate Endometrial Receptivity. G protein-coupled receptor 30(GPR 30) is a new type of ER, researches on which are newly started. And relationship between GPR30 and Endometrial Receptivity is still unknown now. In our study, we studied endometria changes of patients with PCOS in implantation period, discussed the influence factors of Endometrial Receptivity through four aspects as follows, search for the reasons of lower pregnancy rate of PCOS patients, and offer theory proof to treatment of infertility patients with PCOS.There are two parts in the study of the Endometrial Receptivity in the women with PCOS during the implantation period. Part I: Study on Expression of Nucleolar Channel System in Endometrium during Window of Implantation of Women with Polycystic Ovary Syndrome; part II: Study on Expression of GPR30, ERα, ERβand PR in Endometrium during Window of Implantation of Women with Polycystic Ovary Syndrome.Part I Study on Expression of Nucleolar Channel System in Endometrium during Window of Implantation of Women with Polycystic Ovary SyndromeObjectiveTo observe the ultramicrostructure, expression and distribution of endometrial NCS of PCOS patients during the implantation window, and discuss the influencial factors about it.Methods The case-control study was carried out between 15 PCOS patients and 15 control samples. Continuous observed the development of follicles from the 8~10th days of menstruation, performed endometrial biopsies, and measured concentration of sex hormone blood on the day of LH7. All the samples wer performed routine HE staining, and then observed morphous of them by light microscope and SEM. Serum P were measured by chemiluminoimmunoassay (CLIA). Endometrial thickness and pattern was assessed in trans-vaginal ultersonography on ovulation day. At last, discussed the rationships between NCS expression and Progesterone concentration, analyzed the morphology and ultrasonics changes of PCOS patients, which influenced Endometrial Receptivity.Results1 The clinical characteristics of the cases There was no significant difference between this two groups, both in age and body mass index(BMI) (P >0.05). There was significant LH/FSH and T difference between this two groups (P <0.05).2 Morphous of endometriumIn control group,the endometrial glands are twist and abundant, the monolayer glandular epithelium is stylolitic arranged orderly, the glandular cavity is wide and large; The cytoplasm is abundant, and the nucleus is orbicular-ovate and situated in cytoplasmic matrix; There was much acidophilia secretion from endometrial glands; There was abundant vessel dstriuted in the rarefaction stroma. In control group, 11(73.33%) samples were during the midsecretory phase of the menstrual cycle,3 samples during the prosecretory phase and 1 sample during the proliferative phase(26.67%).In PCOS group, the endometrial glands were puny, and little acidophilia secreted from the glands. We observed inflammatory cells scattered in part of the glandular cavity. 5(33.33%) samples were during the midsecretory phase,7 samples during the prosecretory phase and 3 samples during the proliferative phase(66.67%).3 Result of NCS expression in endometrium by TEM We observed endometrium glandular epithelium by TEM. In control group, there were much more longer microvillus on the surface, which always had cytoplasmic process on top of the microvillus. The orbicular-ovate nuclei were located at the bottom, and many NCS were seen in it. The typical appearance of the NCS was always accompanied with Pinopode. NCS may beside or in the nucleolus or nuclear membrane. In difference surface NCS appeared differently, like globosity, bostrychoid, ring or well-arranged net. In suitable surface we observed lumina of NCS was connected with perinuclear cisterna. In cytoplasm, rough endoplasmic reticulum was well development, free ribosomes were abundant and uniform distribution, many megamitochondrion were seen, and multipul glycogen deposited at the top or bottom of the cell. In PCOS group, 7 samples expressed no NCS at all, and NCS were seen by chance in other samples; In control group, typical NCS could be seen frequently. We also observed that NCSs were always accompanied with abundant Pinopode. Because of limited samples and restricted experiment condition, statistical treatment was unavailable, further research was necessary to identify meaning of NCS expression.4 Thickness and pattern of endometrium on ovulation dayThere was no significant thickness difference between the two groups(9.30±1.47 vs 7.89±1.12) mm(P >0.05). The pattern of endometrium in control group: 10 of A; 4 of B and 1 of C; in PCOS group: 5 of A; 7 of B and 3 of C.5 Serum progestin level (ng/ml) in window of implantionSerum progestin of PCOS group was significantly lower than that of the control group(10.81±5.09 vs 16.73±2.59) pg/ml (P<0.05).Summary1 NCS expression of PCOS patients during the window of implantation is lower, accordingly results in descent of Endometrial Receptivity.2 Endometrial pattern of PCOS group is worse than the other group, which may cause lower receptivity and pregnancy rate.3 Decrease of serum progestin probably lead to lower expression of NCS in PCOS patients, thus result in lower pregnancy rate.Part 2 Study on Expression of GPR30, ERα, ERβand PR in Endometrium during Window of Implantation of Women with Polycystic Ovary SyndromeObjectiveTo discuss expression of GPR30, ERα, ERβand PR in Endometrium during Window of Implantation, analyze the rationship among them.MethodsThe samples were achieved like above. GPR30, ERα, ERβand PR were measured by SP immmunohistochemical techniques. Immunostaining outcome was analyzed by the Motic Med 6.0 medical image analysis system.Results1 Expression of GPR30 in endometriumGPR30 was well-distributed in nucleus and cytoplasm of the endometrial glandular epithelial cells, and expressed much more in cytoplasm than in nucleus. The endometrium expressed much less GPR30 in PCOS group than in control group (P <0.05).2 Expression of ERαand ERβin endometriumERαand ERβwas mostly distributed in the nucleus of the endometrial glandular epithelial cells, and sprinkle expressed in the cytoplasm. The endometrium expressed significantly less ERαand ERβin PCOS group than that in control group (P <0.05).3 Expression of PR in endometriumPR was mostly distributed in nucleus of the endometrial glandular epithelial cells, and sprinkle expressed in cytoplasm. The endometrium expressed much more PR in PCOS group than in control group (P <0.05).Summary1 GPR30 was well-distributed in nucleus and cytoplasm of the endometrial glandular epithelial cells, and expressed more in cytoplasm than in nucleus. During window of implantation, GPR30 may participate in quickly transduction of information between the endometrium and the embryo, thus affected implantation of the embryos.2 ERαand ERβwas mostly distributed in nucleus of the endometrial glandular epithelial cells, and lower expressed in PCOS samples, which probably result in descent of the endometrium reaction ability to estrogen and decrease of Endometrial Receptivity.3 PR was mostly distributed in nucleus of the endometrial glandular epithelial cells, and higher expressed in PCOS samples.Conclusions1 During the window of implantation, expression of NCS,GPR30,ERαand ERβin PCOS patients is lower than common people, and expression of PR were higher, accordingly results in descent of Endometrial Receptivity and pregnancy rate.2 It is practicalitical to comprehensively assess Endometrial Receptivity based on morphology,ultrasonic, molecule- bilology and endocrinology.