Polycystic Ovary Syndrome Granulosa Cells Research Articles

MON-232 Differential Regulation of Genes Relevant to Reproductive Function in Human Granulosa Cells of Small Antral Follicles from Women with or without Polycystic Ovary Syndrome

Introduction: Polycystic Ovary Syndrome (PCOS) is the most common endocrinopathy in young women, affecting 5-10% of women of reproductive age. Despite its high prevalence, the underlying pathogenesis of PCOS remains complex and incompletely understood. Aberrant function of granulosa cells has been implicated in the aetiology of PCOS. Material and methods: Granulosa cells (GCs) were obtained from follicular fluid from (unstimulated) individual small antral follicles (SAF) isolated at the time of removal of ovarian tissue for fertility cryopreservation in young women. Granulosa luteal (GL) cells were collected from pooled follicular fluid collected at the time of oocyte retrieval in women undergoing IVF. RNA was extracted and RT-qPCR carried out to analyse expression of genes involved in granulosa cell function, including steroidogenesis and growth. 30 GC samples from individual small antral follicles from 15 women with regular cycles and 31 samples from 10 women with PCOS were included. Mean follicle size was similar between groups (mean 5.5±1mm). Luteinised GCs (GLCs) from 8 women with and without PCOS were also included. Results: 92% of GC samples from SAF expressed LH receptor (LHCGR) RNA. Expression of LHCGR was not significantly higher in PCOS GCs but there was a difference in distribution of results with a subset of PCOS samples (13%) with 4-6 times higher expression than in that of the controls. LHCGR expression was higher in GLCs compared to GCs (P<0.05). FSH receptor (FSHR) expression was significantly lower in PCOS GCs from SAFs (P<0.05), unlike PCOS GLCs, which expressed higher levels of FSHR (p<0.05). FSHR expression was 14 fold higher in GCs compared to GLCs (p<0.0001). There was no correlation between LHCGR, FSHR or follicle size. Androgen receptor (AR) expression was lower in PCOS GCs from SAF (p<0.01) unlike PCOS GLCs, which expressed more AR than GL controls (P<0.05). AR expression correlated with FSHR (p<0.0001) but not LHCGR. CYP11A1 was lower in both PCOS GCs and GLCs (p<0.05) and expression was 50 fold higher in GLCs than GCs (p<0.0001). Expression of 3BHSD2 was reduced in PCOS GCs (p=0.06). There were no significant differences in expression of STAR, CYP19, INHBA or INHBB between control and PCOS in either GCs or GLCs. Conclusions: Granulosa cells from small antral follicles from women with PCOS have a lower expression of FSHR, AR and CYP11A1 and a trend towards higher LHCGR compared to controls. This expression pattern is suggestive of early luteinisation of granulosa cells in unstimulated small antral follicles in PCOS.

Efficacy of electroacupuncture in regulating the imbalance of AMH and FSH to improve follicle development and hyperandrogenism in PCOS rats

Polycystic ovary syndrome (PCOS) is characterized by hyperandrogenism and follicular arrest. These two characteristics may result from an imbalance between anti-Müllerian hormone and follicle stimulating hormone. Electroacupuncture is effective in improving hyperandrogenism and follicular arrest in PCOS; however, the mechanism is not sufficiently clear. This study aimed to elucidate whether electroacupuncture in PCOS is exerted by regulating an imbalance of anti-Müllerian hormone and follicle stimulating hormone. In this study, a rat model of polycystic ovary syndrome was treated with low-frequency electroacupuncture at acupoints (CV-3 and CV-4). To observe the mechanism of electroacupuncture in PCOS, we first observed the estrous cycle. We then observed ovarian morphology by hematoxylin-eosin staining and evaluated levels of testosterone, estradiol, P450arom, follicle stimulating hormone and its receptor, and anti-Müllerian hormone and its receptor by enzyme-linked immunosorbent assay, western blotting, double immunofluorescence assay and real-time PCR. Our results showed that in 80% of rats in the electroacupuncture acupoints group, their estrous cycle recovered, ovarian morphology significantly improved, testosterone level significantly decreased, and levels of estradiol and P450arom significantly increased in peripheral serum after 14 consecutive days of treatment (P < 0.01). The expression of anti-Müllerian hormone and anti-Müllerian hormone type II receptor decreased (P < 0.05), whereas the expression of follicle stimulating hormone receptor increased (P < 0.05). These results indicated that electroacupuncture improved hyperandrogenism and follicular arrest by decreasing the excessive expression of AMH to regulate FSH and AMH imbalance in granulosa cells in PCOS.

MicroRNA-9 affects isolated ovarian granulosa cells proliferation and apoptosis via targeting vitamin D receptor

MicroRNAs (miRNAs or miRs)-9 expression was reported to be upregulated in the follicular fluid of patients with polycystic ovary syndrome (PCOS). However, whether miR-9 affects ovarian dysfunction of PCOS and the related mechanisms are still unclear. Here we detected miR-9 and vitamin D receptor (VDR) expression in women with PCOS and controls, and investigated whether miR-9 affects ovarian granulosa cells (GCs) proliferation and apoptosis by targeting VDR. We found increased miR-9 and decreased VDR in the blood and isolated ovarian GCs of women with PCOS compared with the controls. MiR-9 promoted GCs proliferation and inhibited GCs apoptosis in vitro, and these effects were attenuated by its target VDR. High concentrations of insulin upregulated miR-9 expression in GCs. In conclusion this study firstly proved miR-9 affects ovarian GCs proliferation and apoptosis through targeting VDR. MiR-9 might be a potential molecular target for improving the dysfunction of GCs in PCOS.

Elevation of antimüllerian hormone in women with polycystic ovary syndrome undergoing assisted reproduction: effect of insulin

To measure blood and follicular antimüllerian hormone (AMH) levels in women with polycystic ovary syndrome (PCOS) undergoing assisted reproductive technologies (ART) and to examine the direct action of insulin on AMH expression in human granulosa cells. Prospective clinical and experimental study. University Hospital-based laboratory. Women with (n = 86) and without (n = 172) PCOS in ART. Blood, follicular fluid, and luteinized granulosa cells were collected from PCOS and non-PCOS women in ART. Hormone levels in blood and fluid, and gene expression in granulosa cells. Serum levels of AMH were elevated and inversely correlated with embryo cleavage rate in PCOS women in ART. Significant higher levels of AMH were also found in small and large follicles collected from PCOS women compared with non-PCOS women. Luteinized granulosa cells from PCOS women showed higher expression of AMH and its receptor AMHR2. Direct effect of insulin in increasing the expression of AMH in the isolated luteinized granulosa cells was observed, with the PCOS granulosa cells responding to a high dose of insulin. Cotreatment with AMH attenuated insulin-induced aromatase expression in the luteinized granulosa cells. These results suggest that insulin may contribute to AMH elevation in PCOS and that AMH counteracts insulin-promoted aromatase expression in granulosa cells.

Erythropoietin-producing hepatocellular A7 triggering ovulation indicates a potential beneficial role for polycystic ovary syndrome.

BackgroundThe ovulatory dysfunction mechanisms underlying polycystic ovary syndrome (PCOS) are not completely understood. And the roles of EPHA7 and EPHA7-regulated pathway factors in the pathogenesis of anovulation remain to be elucidated.MethodsWe used human granulosa cells (hGCs) of PCOS and non-PCOS patients to measure EPHA7 and other target gene expressions. We performed in vitro experiments in KGN cells to verify the molecular mechanisms. Additionally, we conducted in vivo loss- and gain-of-function studies using EPHA7 shRNA lentivirus and recombinant EPHA7-Fc protein injection to identify the ovulation effects of EPHA7.FindingsEPHA7 functions as a critically positive upstream factor for the expression of ERK1/2-mediated C/EBPβ. This protein, in turn, induced the expression of KLF4 and then ADAMTS1. Moreover, decreased abundance of EPHA7 was positively correlated with that of its downstream factors in hGCs of PCOS patients. Additionally, a 1-week functional EPHA7 shRNA lentivirus in rat ovaries contributed to decreased numbers of retrieved oocytes, and a 3-week functional lentivirus led to menstrual disorders and morphological polycystic changes in rat ovaries. More importantly, we found that EPHA7 triggered ovulation in rats, and it improved polycystic ovarian changes induced by DHEA in PCOS rats.InterpretationOur findings demonstrate a new role of EPHA7 in PCOS, suggesting that EPHA7 is an effective target for the development of innovative medicines to induce ovulation.FundNational Key Research and Development Program of China, National Natural Science Foundation, Shanghai Municipal Education Commission--Gaofeng Clinical Medicine, and Shanghai Commission of Science and Technology.

PAI-1 in granulosa cells is suppressed directly by statin and indirectly by suppressing TGF-β and TNF-α in mononuclear cells by insulin-sensitizing drugs.

Plasminogen activator inhibitor-1 (PAI-1) is elevated in women with polycystic ovary syndrome (PCOS), but the regulation in granulosa cells (GCs) is unclear. PAI-1 expression in PCOS ovaries was investigated immunohistologically. PAI-1 expressions in HGrC1, a human GC cell line, were investigated at mRNA and activity levels. The expressions of TGF-β and TNF-α in peritoneal fluid mononuclear cells (PFMCs) were measured with quantitative PCR. Little PAI-1 expression is observed in healthy GCs, whereas GCs of PCOS and atretic follicle exhibit distinct expression in vivo. In vitro study using HGrC1 shows that TGF-β and TNF-α increase PAI-1 mRNA and its activity, and both together exhibit a synergistic effect. The expression of PAI-1 mRNA is suppressed by simvastatin. Moreover, insulin-sensitizing drugs (metformin, pioglitazone, and rosiglitazone) suppress LPS-induced TGF-β and TNF-α mRNA expression in PFMC. Statin and insulin-sensitizing drugs may provide a potential therapy for PCOS via down-regulation of PAI-1 expression in GCs and down-regulation of TGF-β and TNF-α expression in PFMC, respectively.

Expression and regulation of CD36 mRNA in granulosa cells and its relation with clinical characteristics of polycystic ovary syndrome

To study the expression of CD36 mRNA in granulosa cells (GCs) of patients with polycystic ovary syndrome (PCOS) and the impact of testosterone, insulin and PPARγ agonist rosiglitazone on GCs. The expression of CD36 mRNA inGCs of patients with PCOS and normal controls were assayed byreal-time PCR.The level of CD36 mRNA after treatment with testosterone, insulin, and rosiglitazone in GCs ofnormal controls were also tested by real-time PCR. (1) The expression of CD36mRNA in the GCs of PCOS was significantly higher than that of the controls (P<0.05). (2) When testosterone concentration was 1 nmol/L, CD36 mRNA increased in the GCs, but there was no significantdifference compared to the blank control, (P>0.05). When testosterone concentration was 10 nmol/L, the expression of CD36 mRNA in the GCs was higher than that in the blank control with significant difference (P<0.05). When insulin concentration was 10 nmol/L, the expression of CD36 mRNA increased but the difference was not statistically significant (P>0.05). When insulin concentration was 100 nmol/L, the expression of CD36mRNA in the GCswas higher than that in the blank control (P=0.05). When rosiglitazone concentration was 1nmol/L, the expression of PPARγ mRNA in GCs were significantly increased compared with the blank control (P<0.05). The expression of CD36 mRNA atrosiglitazone concentrationof 10 nmol/Lwere significantly increased compared to the concentration of 1 nmol/L (P<0.05). High testosterone and insulin induced the expression of CD36 mRNA.Rosiglitazone increased CD36 mRNA in a dose-related manner in GCs.Increased CD36 mRNA in the GCs of PCOS may be related to the clinical characteristics of PCOS.

Association between BMI and gene expression of anti-Müllerian hormone and androgen receptor in human granulosa cells in women with and without polycystic ovary syndrome.

Anti-Müllerian hormone (AMH) is one of the most reliable markers of ovarian reserve. There is evidence which suggests that BMI may be associated with gene expression of AMH, AMH type II receptor (AMHR-II) and androgen receptor (AR) in human granulosa cells (GC) in women with and without polycystic ovary syndrome (PCOS). To investigate the association between BMI and gene expression of AMH, AMHR-II and AR in human GC in women with and without PCOS. In a cross-sectional study, hormonal profiles were measured among 38 patients with PCOS and 38 subjects without PCOS aged 18-40. AMH, AMHR-II and AR mRNA levels were quantified in cumulus GC. Pearson correlation and multiple linear regressions were used to assess the relationships. Quantitative RT-PCR demonstrated that AMH and AMHR-II expression were negatively correlated with BMI (r=-0·39, P<0·001 for AMH and r=-0·49, P<0·001 for AMHR-II), whereas AR expression was positively correlated with BMI (r=0·46, P<0·001). There is a negative association between AMH, AMHR-II expression and BMI, and a positive association between AR expression and BMI in the GC of PCOS and non-PCOS women.

Functional microarray analysis of differentially expressed genes in granulosa cells from women with polycystic ovary syndrome related to MAPK/ERK signaling.

Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women of reproductive age. Although its aetiology and pathogenesis remain unclear, recent studies suggest that the dysfunction of granulosa cells may partly be responsible. This study aimed to use cDNA microarray technology to compare granulosa cell gene expression profiles in women with and without PCOS to identify genes that may be aetiologically implicated in the pathogenesis of PCOS. The study cohort included 12 women undergoing in vitro fertilization, six with PCOS and six without PCOS. Differential gene expression profiles were classified by post-analyses of microarray data, followed by western blot analyses to confirm the microarray data of selected genes. In total, 243 genes were differentially expressed (125 upregulated and 118 downregulated) between the PCOS and non-PCOS granulosa cells. These genes are involved in reproductive system development, amino acid metabolism and cellular development and proliferation. Comparative analysis revealed genes involved in the mitogen-activated protein kinase/extracellular regulated kinase (MAPK/ERK) signaling pathways. Western blot analyses confirmed that mitogen-activated protein kinase kinase kinase 4 and phospho-ERK1/2 were decreased in PCOS granulosa cells. This study identified candidate genes involved in MAPK/ERK signaling pathways that may influence the function of granulosa cells in PCOS.

Increased Expression of Kindlin 2 in Luteinized Granulosa Cells Correlates With Androgen Receptor Level in Patients With Polycystic Ovary Syndrome Having Hyperandrogenemia

Hyperandrogenemia is the leading defect in patients with polycystic ovary syndrome (PCOS) and considered to be involved in the ovulation dysfunction of PCOS. During the process of ovulation, granulosa cells (GCs) undergo epithelial-mesenchymal transition (EMT), and integrin-interacting protein kindlin 2 is a well-known regulator in EMT. Therefore, our objective here was to compare the expression levels of kindlin 2 in luteinized GCs between patients with PCOS and control women and the relationship between kindlin 2 and PCOS pathogenesis. In this study, kindlin 2 expression was significantly increased in luteinized GCs from patients with PCOS, and kindlin 2 could be induced by testosterone both in vitro and in vivo. Meanwhile, kindlin 2 was positively correlated with androgen receptor (AR) in PCOS GCs. Taken together, kindlin 2 may play a role in luteinized GCs, especially in the case of excess androgen. Further studies are required to assess the specific role of kindlin 2 in follicular development and PCOS pathogenesis.

Parallel Variations of Insulin-Like Peptide 3 (INSL3) and Antimüllerian Hormone (AMH) in Women With the Polycystic Ovary Syndrome According to Menstrual Cycle Pattern

Antimüllerian hormone (AMH) and insulin-like factor 3 (INSL3) represent ovarian functional markers of granulosa and theca cells, respectively. We conducted a prospective study to investigate AMH and INSL3 plasma levels in 3 groups of women with polycystic ovary syndrome (PCOS) classified according to menstrual cyclicity pattern and their relationship with ovarian morphology and hormonal levels. AMH and INSL3 were measured in a cohort of 57 patients with PCOS, divided into 3 groups according to menstrual status: eumenorrheic (PCOS-E, n = 15), oligomenorrheic (PCOS-O, n = 25), and amenorrheic (PCOS-A, n = 17). Clinical and endocrine characteristics and ovarian morphology were compared among the groups. Twenty-seven age- and weight-matched women without hyperandrogenism were included as controls. According to the menstrual pattern, the women with PCOS-A and PCOS-O had higher INSL3 levels with respect to the control women (P = .025 and P = .004, respectively) and higher but not significant INSL3 levels compared with those of the women with PCOS-E. AMH levels were significantly higher in women with PCOS-A and PCOS-O with respect to those in women with PCOS-E (P < .001 and P < .001, respectively) and control women (P < .001 and P < .001, respectively). Interestingly, a significant positive correlation was found between INSL3 and AMH blood levels in all women with PCOS (R = 0.43; P = .002) and across the groups (R = 0.41; P < .001). INSL3 and AMH levels are significantly correlated with each other in women with PCOS, and they are significantly increased, particularly in the presence of amenorrhea and oligomenorrhea. INSL3 and AMH may reflect a dysfunction of PCOS thecal and granulosa cells, which are responsible for the increased androgen production and chronic anovulation of this condition.

Differential expression of inflammation-related genes in the ovarian stroma and granulosa cells of PCOS women

Polycystic ovary syndrome (PCOS) is the most common female endocrine disorder. Ovarian changes in PCOS women are well characterized by ultrasound. However, the ovarian pathophysiology is not fully understood. The aim of this study was to characterize the expression, in both the central ovarian stroma and in granulosa cells (GCs), of a number of genes, including several inflammation-related genes, which have been hypothesized to be involved in the pathophysiology of PCOS. Biopsies of the central ovarian stroma were obtained from PCOS women (Rotterdam criteria) and from normally ovulating women in follicular phase. GCs were retrieved from PCOS-women and non-PCOS women, undergoing in vitro maturation. The expressions of 57 genes were analyzed by quantitative-PCR using a low-density-gene array. The main outcome measures were over-expression or under-expression of the specific genes. The results showed that in the central stroma of PCOS ovaries, five inflammation-related genes (CCL2, IL1R1, IL8, NOS2, TIMP1), the leukocyte marker CD45, the inflammation-related transcription factor RUNX2 and the growth factor AREG were under-expressed. The growth factor DUSP12 and the coagulation factor TFPI2 were over-expressed. In the GC of PCOS, all of the differentially expressed genes were over-expressed; the inflammation-related IL1B, IL8, LIF, NOS2 and PTGS2, the coagulation-related F3 and THBS1, the growth factors BMP6 and DUSP12, the permeability-related AQ3 and the growth-arrest-related GADD45A. In conclusion, the results indicate major alterations in the local ovarian immune system of PCOS ovaries. This may have implications for the PCOS-related defects in the inflammation-like ovulatory process and for the susceptibility to acquire the inflammatory state of ovarian hyperstimulation syndrome.

Uncoupling protein 2 expression affects androgen synthesis in polycystic ovary syndrome

The roles of uncoupling protein-2 (UCP2) on the androgen synthesis of granulosa cells derived from patients with polycystic ovary syndrome (PCOS) and normal subjects were explored. Primary human granulosa cells from 18 patients who received in vitro fertilization (IVF) were examined; nine patients had PCOS with hyperandrogenism. Primary cultures were treated with genipin, a proton leak inhibitor, guanosine diphosphate (GDP), an UCP inhibitor, and triiodothyronine (T3), an inducer of UCP gene expression. Mitochondrial membrane potential was determined using the JC-1 assay. T3 induced P450scc and UCP2 expressions and testosterone synthesis in both normal and PCOS granulosa cells. Their expressions in response to T3 treatments were correlated in the PCOS group. Differences in testosterone synthesis were observed between normal and PCOS cells in response to genipin. Increased mitochondrial membrane potential was observed in response to genipin and GDP; while T3 decreased it. Increased ovarian UCP2 expression in response to T3 treatment in PCOS may alter pregnenolone synthesis by influencing P450scc expression, thus altering testosterone production. Further in vivo studies are necessary to fully elucidate the role of UCP2 in the hyperandrogenism commonly observed in PCOS.

Anti-mullerian hormone is elevated in women with polycystic ovary syndrome but is unaltered following hCG stimulation

Polycystic ovary syndrome (PCOS) is characterized by increased luteinizing hormone (LH) secretion, driving excess androgen production. Recent studies show that anti-mullerian hormone (AMH) production is also elevated in PCOS and serum AMH levels positively correlate with LH and androgen levels. In vitro data suggest a causal relationship: Stimulation with LH significantly increases AMH secretion from granulosa cells of PCOS but not of normal women. We investigate the possibility of a direct effect of LH on AMH production in vivo by assessing AMH and androgen responses after human chorionic gonadotropin (hCG) stimulation (as a surrogate for LH) in PCOS and normal women.

The mechanism of mTOR (mammalian target of rapamycin) in a mouse model of polycystic ovary syndrome (PCOS)

Polycystic ovary syndrome (PCOS) is a common and complex endocrine disorder affecting 5-10% of women in reproductive age that is characterized by hyperandrogenism, oligo- or anovulation and infertility. However the pathophysiology of PCOS still remains unknown. The mammalian target of rapamycin (mTOR) is a central component that regulates various processes including cell growth, proliferation, metabolism, and angiogenesis. mTOR signaling cascade has recently been examined in ovarian follicles where it regulates granulosa cell proliferation and differentiation. mTOR functions as two complexes, mTOR complex 1 and 2. Therefore, we hypothesized that mTORC1 and/or 2 may have important role in proliferation of theca and granulosa cells in PCOS. In the present study, we sought to determine the mTOR signaling pathway in PCOS mouse ovary. We designed 3 groups: Control (C, no treatment), PCOS (P, The injection of DHEA (6 mg/100 g BW in 0.1 ml of sesame oil) (s.c) for 20 consecutive days), Vehicle (V, daily (s.c) sesame oil alone injection). Our results showed that mTORC1 and mTORC2-mediated signaling may play a role in PCOS mouse ovary. These findings provide evidence that mTORC1 and mTORC2 may have responsibility in increased ovarian follicular cell proliferation and growth in PCOS. Consequently, these results suggest that the mTOR signaling pathways (mTORC1 and mTORC 2) may create new clinical strategies to optimize developmental competence of PCOS should target correction of the entire follicle growth, oocyte development process and anovulatory infertility in PCOS.

Expression and regulation of adipocyte fatty acid binding protein in granulosa cells and its relation with clinical characteristics of polycystic ovary syndrome

Increased expression of adipocyte fatty acid binding protein (FABP4) is associated with type 2 diabetic, high triglycerides, increased lipid peroxidation, and inflammation markers. To study the expression of FABP4 mRNA in granulosa cells of patients with polycystic ovary syndrome (PCOS) and the impact of testosterone, insulin, and PPARγ agonist rosiglitazone on granulosa cells (GCs), and to investigate the relationship of serum FABP4 levels with clinical characteristics in patients with PCOS. The expression of FABP4 mRNA in GCs of patients with PCOS and normal controls were assayed by RT-PCR. We assessed the level of FABP4 mRNA after treatment with testosterone, insulin, and rosiglitazone in GCs from normal controls. Serum FABP4 were assayed from 96 patients with PCOS (obese and nonobese 48 cases, respectively) and 80 healthy normal controls (obese and the nonobese 40 cases, respectively). The expression of FABP4 mRNA was higher in the GCs of PCOS than that of the controls (P<0.05). FABP4 mRNA expression was up-regulated by testosterone, insulin, and rosiglitazone at different dosages. Serum FABP4 levels were higher in the nonobese PCOS group than that of the nonobese controls (8.9±5.1 ng/ml vs. 4.8±0.7 ng/ml), and in the obese PCOS group than that of the obese controls (28.2±14.0 ng/ml vs. 15.6±6.6 ng/ml), respectively (P<0.05). Multiple linear regression analyses showed that serum FABP4 level was independently associated with HOMA-IR, BMI, and testosterone (P<0.05). Increased FABP4 was related to the clinical characteristics of PCOS.

Follicle-stimulating hormone suppressed excessive production of antimullerian hormone caused by abnormally enhanced promoter activity in polycystic ovary syndrome granulosa cells

To explore: 1) the cause of excessive production of antimullerian hormone (AMH); 2) the effect of FSH on overproduction of AMH; and 3) expression of AMH receptor II (AMHRII) in polycystic ovary syndrome (PCOS) granulosa cells. Case-control study. University affiliated hospital. MATERIAL(S): The granulosa cells from 8-10-mm follicles were divided into four groups: unstimulated PCOS granulosa cells (n=12), exogenous FSH (5 ng/mL)-stimulated PCOS granulosa cells (n=12), internal FSH (150-300 IU/d)-stimulated PCOS granulosa cells (n=20), and control granulosa cells from non-PCOS infertile patients with regular menstruation (control; n=29). Exogenous FSH was added into the medium of unstimulated PCOS granulosa cells. The secretory levels and mRNA abundance of AMH, the mRNA and protein levels of AMHRII, and the luciferase activity of AMH. Both the secretiory level and mRNA abundance of AMH were significantly enhanced in PCOS granulosa cells. They could be partially suppressed by FSH. The mRNA and protein levels of AMHRII in PCOS granulosa cells were significantly increased. However, they were not affected by FSH. The luciferase activity of AMH in PCOS granulosa cells was significantly amplified but could be suppressed by FSH. Enhanced promoter activity can cause excessive production of AMH in PCOS granulosa cells. FSH may inhibit the excessive secretion of AMH by suppressing the luciferase activity of AMH promoter, but it has no effect on AMHRII expression.

Different phenotypes of polycystic ovary syndrome by Rotterdam criteria are differently steroidogenic but similarly insulin resistant

In traditional Chinese medicine, polycystic ovary syndrome (PCOS) is considered an anovulation disorder related to ovarian insulin resistance. The three phenotypes of PCOS, according to the Rotterdam criteria, are differently steroidogenic but similarly insulin resistant, suggesting a similar involvement of insulin resistance/hyperinsulinemia in different compartments of the PCOS ovary, namely, overactive theca and/or granulosa cells.

Comparison of Follicle-Stimulating-Hormone-Stimulated Dimeric Inhibin and Estradiol Responses as Indicators of Granulosa Cell Function in Polycystic Ovary Syndrome and Normal Women

Follicular phase secretion of inhibin B, like that of estradiol (E(2)), correlates with the quantity and quality of developing follicles. However, it has not been established whether inhibin B responses to gonadotropin stimulation parallel those of E(2) as a reflection of granulosa cell functional capacity. Our objective was to determine whether inhibin B responses to FSH stimulation are similar to those of E(2) in women with polycystic ovary syndrome (PCOS) and normal women. We conducted a prospective study to compare ovarian responses in two groups of women at a general clinical research center in a tertiary academic medical center. Women with PCOS, 18-35 yr (n = 19), and normal ovulatory controls, 18-35 yr (n = 7), were recruited for study. Serum samples were measured over a 24-h period after an iv injection of recombinant human FSH, 150 IU. Serum E(2), inhibin A, and inhibin B responses after FSH administration were assessed. In PCOS women, the 24-h production of inhibin B and E(2) after FSH was significantly greater than that of normal controls. Within the PCOS group, the fold change in inhibin B was significantly greater than that of E(2). Inhibin A responses between groups were similar and of markedly lower magnitude. FSH-stimulated inhibin B responses may be employed to assess the functional capacity of granulosa cells in PCOS and normal women.

Impaired insulin-dependent glucose metabolism in granulosa-lutein cells from anovulatory women with polycystic ovaries

Insulin resistance and hyperinsulinaemia are well-recognized characteristics of anovulatory women with polycystic ovary syndrome (PCOS) but, paradoxically, steroidogenesis by PCOS granulosa cells remains responsive to insulin. The hypothesis to be tested in this study is that insulin resistance in the ovary is confined to the metabolic effects of insulin (i.e. glucose uptake and metabolism), whereas the steroidogenic action of insulin remains intact. Granulosa-lutein cells were obtained during IVF cycles from seven women with normal ovaries, six ovulatory women with PCO (ovPCO) and seven anovulatory women with PCO (anovPCO). Mean body mass index was in the normal range in all three groups. Granulosa-lutein cells were cultured with insulin (1, 10, 100 and 1000 ng/ml) and LH (1, 2.5 and 5 ng/ml). Media were sampled at 24 and 48 h and analysed for glucose uptake, lactate production and (48 h only) progesterone production. Insulin-stimulated glucose uptake by cells from anovPCO was attenuated at higher doses of insulin (100 and 1000 ng/ml) compared with that by cells from either ovPCO (P=0.02) or controls (P=0.02). Insulin and LH stimulated lactate production in a dose-dependent manner, but insulin-dependent lactate production was markedly impaired in granulosa-lutein cells from anovPCO compared with either normal (P=0.002) or ovPCO (P<0.0001). By contrast, there was no difference in insulin-stimulated progesterone production between granulosa-lutein cells from the three ovarian types. Granulosa-lutein cells from women with anovPCOS are relatively resistant to the effects of insulin-stimulated glucose uptake and utilization compared with those from normal and ovPCO, whilst maintaining normal steroidogenic output in response to physiological doses of insulin. These studies support the probability of a post-receptor, signalling pathway-specific impairment of insulin action in PCOS.