| Polycystic ovary syndrome(PCOS) is a kind of endocrine disorder disease associated with reproductive dysfunction in reproductive women. Yet, its etiology and pathology mechanism has not been fully studied. In recent years, researches have shown that PCOS might originates from oocytes and the abnormal endocrine in PCOS patients may be motivated by ovaries. As two oocyte growth factors, growth differentiation factor 9(GDF-9) and bone morphogenetic protein 15(BMP-15) are thought to be crucial for follicular regulation and development of oocytes, as important for follicle environment stability and reproductive function. There have been a few studies showing that GDF-9, BMP-15 may be valuably involved in the follicular dysplasia of PCOS and that GDF-9, BMP-15 may participate in the pathophysiology of ovarian response. Therefore, this study will explore the expression of oocytes derived growth factors in PCOS patienes in ontrolled ovarian hyperstimulation(COH) cycles and their effects on ovarian response and in vitro fertilization and embryo transfer(IVF-ET) outcomes, to research the possible mechanism of ovarian regulation, finally to provide further theoretical foundation for improving the reproductive function of infertile patients with PCOS.Object:To detect the expression levels of GDF-9 and BMP-15 in granulosa cells and follicular fluid in PCOS patients, analyze the discrepancy of GDF-9, BMP-15 expression levels between PCOS and control group during COH cycles, and to explore the effects of GDF-9, BMP-15 expression levels on follicle development, ovarian response and IVF-ET outcomes in PCOS patient.Methods:75 PCOS patients and 150 infertile cases due to tubal factors who underwent IVF-ET at the same period were recruited as the experimental group and control group. Granulosa cells and follicular fluid were collected after oocyte retrieval followed by ontrolled ovarian hyperstimulation. Immunocytochemistry was used to detect localization and expression of GDF-9, BMP-15 in granulosa cells. Real-time PCR and western blot were used to detect the expression levels of GDF-9, BMP-15 mRNA in granulosa cells and their proteins in follicular fluid, respectively. Pearson analysis and logistic regression analysis were used to analyze the correlation of GDF-9, BMP-15 expression levels with ovarian response and IVF-ET outcomes. Receiver operating characteristic curve was analyzed to evaluate the accuracy of GDF-9, BMP-15 in predicting clinical pregnancyResults:1. The constituent ratio of ovarian response were no statistically different between PCOS group who had 25 higher ovarion responders, 38 normal ovarian responders, 12 lower ovarian responders and control group who had 43, 77 and 30, respectively. Oocyte retrieved rate, oocyte maturation rate and normal cleavage rate in PCOS group were significantly lower than that of control group and the transplanted cancellation rate were significantly higher(P<0.05). Implatation rate, clinical pregnancy rate and early abortion rate between the two group were no statistically different(P>0.05).2. Both GDF-9 and BMP-15 proteins were detected in the cytoplasm of granulosa cells in the two groups by immunocytochemistry analysis. GDF-9, BMP-15 mRNA expression levels in granulosa cells and their proteins levels in follicular fluid of PCOS group were significantly lower than those of control group(P<0.01).3. GDF-9 mRNA levels in granulosa cells and its proteins levels in follicular fluid were significantly different among the three ovarian response subgroups in PCOS patients, which were highest in high ovarian responders and lowest in poor ovarian responders(P<0.05); the expression levels of BMP-15 mRNA in granulosa cells and its proteins in follicular fluid were no significantly different among the three subgroups(P>0.05). In contro group GDF-9 mRNA levels in granulosa cells and its proteins levels in follicular fluid were significantly different among the three ovarian response subgroups, which were highest in high ovarian responders and lowest in poor ovarian responders(P<0.05); the expression levels of BMP-15 mRNA in granulosa cells and its proteins in follicular fluid of poor ovarian responders twere significantly higher han that of other two subgroups(P<0.05).Pearson analysis showed that the no. of retrieved oocytes were positively correlated with antral follicle cout(AFC), basal LH, GDF-9 mRNA level in granulosa cells and GDF-9 protein level in follicular fluid(P<0.05), but negatively correlated with age,dose of Gn, basal FSH and BMP-15 protein level in follicular fluid(P<0.05). Logistic regression showed that AFC, GDF-9 mRNA level in granulosa cells and GDF-9 protein level in follicular fluid were the main influencing factors of high ovarian response and that age, BMP-15 mRNA level in granulosa cells and its protein level in follicular fluid were the main influencing factors of poor ovarian response.4. There were 19 cases of clinical pregnancy and 25 cases of no-pregnancy in PCOS group, while in control group were 50 cases and 59 case, respectively. GDF-9, BMP-15 mRNA levelsin granulosa cells and proteins levels of clinical pregnancy group were significantly higher than that of no-pregnancy group in both PCOS and control groups(P<0.05).Pearson analysis showed that oocyte maturation rate was positively correlated with GDF-9 mRNA level in granulosa cells, GDF-9 protein level and BMP-15 protein level in follicular fluid and that no. of high-quality embryos was negatively correlated with age, positively correlated with GDF-9 protein level in follicular fluid. Receiver operating characteristic curve discovered that both GDF-9, BMP-15 mRNA in granulosa cells and their proteins in follicular fluid can predict clinic pregnency(AUC were 0.667, 0.646, 0.836, 0.807, respectively, P<0.05).Conclusions:1. The oocyte retrieved rate, oocyte maturation rate and normal cleavage rate in PCOS group were significantly lower while expression levels of GDF-9 and BMP-15 mRNA in granulosa cells and their proteins in follicular fluid were were significantly lower. GDF-9, BMP-15 may valuably involved in the follicular dysplasia of PCOS.2. GDF-9, BMP-15 took part in the pathophysiology of ovarian response in PCOS patients. Keeping a moderate amout of GDF-9, BMP-15 expression levels may be a target to control ovarian response in PCOS patients.3. Higher expression levels of GDF-9,BMP-15 predicted better pregnancy outcome. The expression levels of GDF-9,BMP-15 proteins in follicular fluid will valuably predict clinical pregnancy rate. |
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