Chk1/2 inhibitor AZD7762 blocks the growth of preantral follicles by inducing apoptosis, suppressing proliferation, and interfering with the cell cycle in granulosa cells.

BACKGROUND: Checkpoint kinases 1/2 (Chk1/2) have an important role in somatic cell development and oocyte meiotic maturation. However, the role of Chk1/2 in folliculo-genesis has not been fully elucidated. The aim of this study was to assess the effects of Chk1/2 inhibition on ovarian folliculogenesis and granulosa cell development in mice. METHODS: Preantral follicles (100-120 µm) and granulosa cells from pre-ovulatory follicles (pre-GCs) of mice were isolated and cultured with or without Chk1/2 inhibitor AZD7762. Preantral follicles were cultured for 96h. Then, follicle morphology and fol-licular growth were assessed every 48h. Granulosa cells were cultured for 48h with or without AZD7762, after which cell apoptosis, cell proliferation, and cell cycle analysis were assessed; meanwhile, the mRNA expression of PCNA and Bax were measured by real-time RT-PCR, and PCNA and Bax protein were measured by Western blot. RESULTS: Compared with control follicles, AZD7762 inhibited growth of preantral folli-cles (P<0.05). Furthermore, inhibition of Chk1/2 significantly induced apoptosis (P<0.05) and inhibited the proliferation of granulosa cells (P<0.01), arrested cell cycle at S and G2/M phases, and decreased G1 phase fraction (P<0.001). Also, the expres-sion of PCNA mRNA and protein were reduced (P<0.01), while Bax mRNA and pro-tein were increased (P<0.05) post AZD7762 treatment in granulosa cells. CONCLUSIONS: This study revealed that Chk1 and Chk2 have a crucial role during preantral follicular development by regulating the proliferation and apoptosis of granu-losa cells.

Knockdown of Chk1 inhibits proliferation and promotes apoptosis in mouse granulosa cells and its regulation mechanism by miR-15a and miR-16.

Checkpoint kinase 1 (Chk1) is a protein kinase which preserves the genome integrity, and works as an evolutionally conserved DNA damage response and cell cycle checkpoint. However, the functional roles and regulatory mechanism of Chk1 in mouse granulosa cells (GCs) have not been fully elucidated. In this study, by RNA interfering, Chk1 gene was knocked down in GCs. Knockdown of Chk1 inhibited proliferation and increased apoptosis of GCs (p < 0.05), respectively; in addition, cell cycle of GCs was arrested at S and G2/M phases. Further qRT-PCR results showed that cell cycle factors (Cyclin B1 and Cyclin D 1) and a marker gene of proliferation (PCNA) were downregulated (p < 0.001), while apoptotic factors (p53b, p21, caspase-3, and Bax) were upregulated (p < 0.01), which suggested that knockdown of Chk1 may inhibit proliferation, regulate cell cycle, and promote apoptosis at the transcriptional level in GCs. In vitro studies showed a negative correlation between Chk1 mRNA and miR-16 expression during follicular development. To elucidate the relationship between Chk1 and miR-15a/16, luciferase reporter plasmids were constructed and luciferase assays revealed that both miR-15a and miR-16 could bind to the 3' UTR of Chk1 mRNA, and significantly downregulate the protein level of Chk1 (p < 0.01), while miR-16, not miR-15a, could significantly decrease the mRNA level of Chk1 (p < 0.05). This result indicated that miR-16 directly induced Chk1 mRNA destabilization, while miR-15a regulated Chk1 expression through translational repression. Taken together, this study uncovered the roles of Chk1 in mouse granulosa cells and its regulation by miR-15a and miR-16 through different mechanisms.

Comparison of Pregnancy Outcomes Between Single-Morula Embryo Transfer and Single-Blastocyst Transfer in Fresh IVF/ICSI Cycles.

BACKGROUND This study investigated the effectiveness and feasibility of day 4 (D4) morula embryo transfer (ET) in comparison with day 5 (D5) blastocyst ET, with regards to their clinical data, laboratory test results, and pregnancy outcomes. MATERIAL AND METHODS This retrospective cohort study enrolled 1070 patients, including 178 cases in group D4 and 892 cases in group D5. The endpoint was live birth rate after fresh embryo transfer. Furthermore, the clinical outcomes of D4 embryos with different morphology were compared and assigned to 3 groups: in group 1 (n=66) the embryos were compacted but not expanded, in group 2 (n=102) the embryos were compacted and expanded (early blastocyst), and in group 3 (n=10) the embryos were not compacted. RESULTS Groups D4 and D5 had comparable clinical pregnancy rates (53.37% vs. 59.97%) and live birth rates (43.25% vs 50.89%), and there were no significant differences between the 2 groups. In group 3, there was only 1 clinical pregnancy and no live birth. In comparison between group 1 and group 2, the clinical pregnancy rate of group 2 showed an upward trend (48.48% vs 60.78%), but there was no significant difference. There was also no statistically significant difference in the live birth rate between the 2 groups (42.42% vs 49.01%). CONCLUSIONS Transferring of compacted embryos or early blastocysts can result in high clinical pregnancy rates and live birth rates. In addition to the cleavage and blastocyst ET, morula ET may serve as an alternative option for the clinician.

[Comparison of the efficiency between in-vitro maturation and in-vitro fertilization after early follicular phase GnRH agonist down-regulation in infertile women with polycystic ovary syndrome].

OBJECTIVE: To compare the outcomes of in-vitro maturation (IVM) and in-vitro fertilization (IVF) after early follicular phase gonadotropin-releasing hormone agonist (GnRH-a) down-regulation in infertile patients with polycystic ovary syndrome (PCOS). METHODS: From July 2010 to December 2012, 72 infertile patients with PCOS undergoing assisted reproductive technology treatment in the Affiliated First Hospital of Wenzhou Medical University were enrolled in this study. The patients were divided into 2 groups, which were patients with early follicular phase down-regulation IVM (36 cases) at IVM group and early follicular phase down-regulation long protocol IVF (36 cases) at IVF group. The laboratory parameters and clinical outcomes were compared between two groups. RESULTS: (1) Lab parameters: a total of 442 oocytes were retrieved in group IVM, and 560 were in group IVF. The rate of mature oocytes of 83.8% (469/560) and high-quality embryos of 70.9% (212/299) at group IVF were significantly higher than that of group IVM[54.1% (239/442) and 50.7% (73/144), retrospectively, P < 0.01]. In group IVM, the average duration of gonadotropin (Gn) was (2.8 ± 1.5) days and the average dosage of Gn was (285 ± 169) U, which were significantly lower than (11.0 ± 1.0) days and (1499 ± 165) U in group IVF (P < 0.01). The mean number of oocytes retrieved 12.8 ± 2.5, fertilization rate of 64.8% (155/239), and implantation rate of 31% (23/74) in group IVM and 15.6 ± 3.1, 65.5% (307/469), 31% (23/74) in group IVF, which did not reach statistical difference (P > 0.05) . (2) Clinical outcomes: the clinical pregnancy rate (17/31, 55%) of IVF group was not significantly higher than that 44% (14/32) at IVM group (P > 0.05). The abortion rate was 1/17 at Group IVF and 1/14 in group IVM, which did not show statistical difference. Women at IVM group has no ovarian hyper-stimulation syndrome (OHSS) cycle, group IVF has 31% (11/36) cycles presented moderate and severe OHSS. CONCLUSIONS: Infertile patients with PCOS undergoing IVM and IVF treatment after early follicular phase GnRH-a down-regulation can get satisfactory laboratory and clinical outcome. In addition to short treatment cycle, IVM can also avoid the occurrence of OHSS completely, but it has a rising trend in the abortion rate. IVF has a high incidence of OHSS, meanwhile, it increases the dosage of gonadotropins.

[Study on clinical effect on infertility women with polycystic ovary syndrome treated by in vitro maturation and in vitro fertilization-embryo transfer].

OBJECTIVE: To compare clinical and laboratory outcomes of in vitro maturation (IVM) with in vitro fertilization-embryo transfer (IVF-ET) in treatment of infertility associated with polycystic ovary syndrome (PCOS). METHODS: From Jan.2007 to Dec.2010, infertile patients with PCOS underwent 701 cycles in First Affiliated Hospital of Wenzhou Medical College were studied retrospectively. Those were divided into 293 cycles of IVM group and 408 cycles of IVF/intra-cytoplasmic sperm injection (ICSI) group. The average transplantation rate, mean number of retrieval oocytes, maturation rate, fertilization rate, cleavage rate, high quality embryo rate, embryo implantation rate, pregnancy rate per transfer, pregnancy outcomes and incidence of ovarian hyperstimulation syndrome (OHSS) of the two methods of treatment were compared between two groups. RESULTS: There were 275 cycles in IVM group and 342 cycles in IVF/ICSI group established embryo transfer. The transplantation rate was 93.9% (275/293) in IVM group and 83.8% (342/408) in IVF/ICSI, which reached statistical difference (P < 0.01). The maturation rate of 56.64%, cleavage rate of 88.08%, high quality embryo rate of 38.72% and embryo implantation rate of 17.8% in IVM group were significantly lower than 91.09%, 94.91%, 51.50% and 25.4% in IVF/ICSI group (all P < 0.01). The clinical pregnancy rate per transfer were 37.8% (104/275) in IVM group and 44.2% (151/342) in IVF/ICSI group, which did not show statistical difference (P > 0.05). The mean number of oocytes (12.9 ± 6.5 vs. 12.9 ± 7.9) and fertilization rate (76.52% vs. 70.75%) didn't show significant difference between IVM group and IVF/ICSI group (P > 0.05). The 21.3% (87/408) cycles presented mild to moderate OHSS and 2.0% cycles (8/408) presented severe OHSS in IVF/ICSI group. While, no OHSS cycles were observed in IVM group. CONCLUSION: IVM could get similar clinical pregnancy rates compared with IVF/ICSI in patient with PCOS, however, it can avoid occurrence of OHSS.

Effects of oral contraceptives and metformin on the outcome of in vitro maturation in infertile women with polycystic ovary syndrome.

UNLABELLED: Abstract Objective: To evaluate the effects of oral contraceptives and metformin on the outcome of in vitro maturation (IVM) in infertile women with polycystic ovary syndrome (PCOS). METHODS: This is a retrospective study of 108 women with PCOS, subject to 152 cycles of IVM treatment. The study was held at the Reproductive Medicine Center of the First Affiliated Hospital of Wenzhou Medical College, People's Republic of China. Before entering IVM treatment, 54 patients who received oral contraceptive pill (marvelon, 0.15 mg desogestrel, and 0.03 mg ethinylestradiol), one tablet every day, and metformin 500 mg twice or three times per day were defined as the pretreated group, and another 64 patients who were not administered any drugs as the control group. The main outcome measures were the rates of oocyte maturation, fertilization, cleavage, miscarriage, clinical pregnancy, and live birth. RESULTS: There were no significant differences between the two groups in the rates of oocyte maturation, fertilization, cleavage, and clinical pregnancy (p > 0.05). A significantly lower miscarriage rate was obtained in the pretreated group than in the control group (16.13% vs 4.0%, p < 0.01). The live birth rate per embryo transfer seemed to be higher in the pretreated group than in the control group (37.70% vs 30.38%, p = 0.363), but was not statistically significant. CONCLUSIONS: Pretreatment with oral contraceptives and metformin improved the outcome of IVM related to the miscarriage rate and possibly also live birth rate.

[Outcome of pregnancy in women with polycystic ovary syndrome treated by in vitro maturation of immature oocytes].

OBJECTIVE: To investigate pregnancy outcome of infertility women with polycystic ovary syndrome (PCOS) treated by in vitro maturation (IVM) of immature oocytes. METHODS: From Nov. 2003 to Oct. 2007, medical documents of 118 women with PCOS underwent 140 IVM treatment cycles in Reproductive Medical Center of First Affiliated Hospital of Wenzhou Medical College were collected. Follow up of 62 pregnancies were performed by prenatal examination in hospital or telephone query to record perinatal monitoring and pregnancy outcome. RESULTS: There are 62 pregnant women including 5 biochemical pregnancies and 57 clinical pregnancies obtained in 140 transferred cycles, resulting in the pregnancy rate of 40.7% (57/140). The rates of singleton pregnancies, twin pregnancies and triplet pregnancies were 61.4% (35/57), 29.8% (17/57) and 5.3% (3/57), respectively. The rate of ectopic pregnancy was 3.5% (2/57). Seven (7/57, 12.3%) women underwent early abortion during 7 -14 weeks of pregnancy, and 1 case (1/57, 1.75%) with premature rupture of membranes occurred at 22 gestational week. One woman with twin pregnancy spontaneously reduced to singleton at 8 gestational weeks. Totally, the rate of pregnancy complications was 26.3% (15/57) including premature rupture of membranes (1 case), placenta previa (1 case), hypertensive disorder (1 case), preterm delivery (10 cases) and gestational diabetes mellitus (2 cases). Until now 47 women gave birth to 65 infants including 29 singleton infants and 18 twins. One female preterm neonate died after 6 days' delivery due to pneumonia, no malformation was observed on the other neonates. 21.3% (10/47) of deliveries were premature,76.6% (36/47) of deliveries were full-term, 2.1% (1/47) of deliveries were postterm. The mean birth weight was 2972 gram. The rate of infants with low weight was 26.2% (17/65). CONCLUSIONS: A relatively high clinical pregnancy rate has been achieved, the rates of early abortion, ectopic pregnancy, pregnancy complications, perinatal mortality, and neonatal malformation occurring after the treatment of IVM in women with PCOS are not mounting. However, the relative high rates of multiple pregnancies, low birth weight and preterm labor were increased.

In vitro maturation and fertilization of oocytes from unstimulated ovaries in infertile women with polycystic ovary syndrome.

OBJECTIVE: To evaluate the effects of in vitro maturation and fertilization of oocytes from unstimulated ovaries in infertile women with polycystic ovary syndrome (PCOS). DESIGN: Retrospective study. SETTING: Reproductive Medicine Center, First Affiliated Hospital of Wenzhou Medical College, People's Republic of China. PATIENT(S): One hundred eighteen women with PCOS undergoing 152 cycles of in vitro maturation treatment. INTERVENTION(S): Oocyte retrieval was carried out by ultrasound-guided puncture on days 9-14 of the cycle. The oocytes were cultured in vitro using maturation culture medium, which consisted of M-199 + 20% fetal bovine serum (FBS) + 75 mIU/mL recombinant FSH +/- 0.5 IU/mL hCG. After the oocytes had matured in vitro, fertilization and embryo transfer were performed. MAIN OUTCOME MEASURE(S): Rates of clinical pregnancy, multiple pregnancies, and live birth. RESULT(S): Relatively optimal laboratory results were obtained in this study. Embryo transfer was performed in 140 cycles, with a clinical pregnancy rate (PR) of 40.0% per transfer. Fifty-six babies have been born and there are 10 ongoing pregnancies. The overall multiple PR was 33.93%. CONCLUSION(S): Our results show that using in vitro matured oocytes from unstimulated ovaries could be offered as an alternative to conventional IVF in women with PCOS, and future work should address ways to decrease the incidence of multiple pregnancies.

[Clinical study on in vitro maturation of immature oocytes transferred from conventional in vitro fertilization.].

OBJECTIVE: To investigate clinical effect of in vitro maturation (IVM) of immature oocytes transferred from conventional in vitro fertilization embryo transfer (IVF-ET) cycles. METHODS: From January 2008 to June 2009, medical documents of 155 infertile patients underwent IVF-ET in the Reproductive Medical Center of First Affiliated Hospital of Wenzhou Medical College were analyzed retrospectively. If more than 20 oocytes were monitored after 5 - 7 days of ovulation induction or follicular developmental retardation were confirmed after 8 - 13 days of ovulation induction, according to patients' wish, IVM were transferred in 60 cycles (group A). In the mean time, IVF was continued in 95 cycles (group B). The mean dosage of gonadotropin, the cancellation rate of cycles, the mean numbers of oocytes retrieved and maturation, the rate of fertilization and excellent quality embryos, pregnancy outcome and the incidence rate of ovarian hyperstimulation syndrome (OHSS) were compared and analyzed. RESULTS: The rates of embryo transfer were 92% (55/60) in group A and 63% (60/95) in group B, which showed significant differences (P < 0.05). In group A, the mean dosage of the gonadotropin, the mean number of oocytes retrieved, the cleavage rate and OHSS rate were (1030 +/- 468) U, 10 +/- 6, 82.2% (231/281) and 0, and were (1544 +/- 338) U, 14 +/- 4, 94.0% (502/534) and 35% (21/60) in group B, respectively, all data above exhibited statistical difference between two groups (P < 0.05). However, the rates of fertilization and excellent quality embryos had no significant differences between two groups (P > 0.05). In group A, the rate of clinical pregnancy per transfer was 53% (29/55) and multiple pregnancy was 14% (4/29), and were 47% (28/60) and 32% (9/28) in group B, they all had no significant differences (P > 0.05). CONCLUSION: IVM of immature oocytes used in conventional IVF cycles not only obtained a high clinical pregnancy rate, but also reduced gonadotropin using dosage and avoided OHSS completely.

[Effect of desogestrel and ethinyl estradiol pretreatment in superovulation cycles with short protocol].

OBJECTIVE: To study the effect of desogestrel and ethinyl estradiol (DEE) pre-treatment combined with gonadotropin releasing hormone agonist (GnRH-a) stimulation in in vitro fertilization-embryo transplantation (IVF-ET). METHODS: A retrospective analysis was performed in 101 infertile women who received a short protocol of GnRH-a for IVF-ET treatment from June 2004 to June 2007 in the Reproductive Medicine Center of First Affiliated Hospital of Wenzhou Medical College. Patients had been pre-treated with oral contraceptive pill (OCP) for two months before GnRH-a combined with recombinant follicle stimulation hormone (r-FSH) treatment (study group, n = 42) or had not been pretreated with OCP (control group, n = 59). A statistical analysis of two groups was carried out for the assessment of ovulation stimulating effect of OCP and its influence on the IVF. RESULTS: Serum FSH was significantly decreased after OCP in the study group. Twelve pregnancies were obtained including 1 case of spontaneous abortion at 7 weeks in the study group, and 11 pregnancies were obtained including 2 cases of spontaneous abortion during 7 -9 weeks in control group. The clinical pregnancy rates in the study group (23%, 12/53) was higher than that in the control group (17%, 11/63), but the differences were not significant (P > 0.05). The miscarriage rate in the study group (8%, 1/12) was lower than that in the control group (18%, 2/11), however no significant differences were found between them (P > 0.05). The cycle cancellation rate in patients of the study group (5%, 3/56)was significantly lower than that in patients of the control group (17%, 13/76, P < 0.05). The differences between patients of the two groups with respect to age, basal level of FSH and luteinizing hormone (LH), antral follicle counts, the mean number of oocyte retrieval, the days of stimulation, total dose of r-FSH used, fertilization rate and embryo cleavage rate, however were insignificant. CONCLUSION: OCP pretreatment combined with short protocol of GnRH-a stimulation in IVF could significantly decrease the cycle cancellation rate, with a declining miscarriage rate and increasing pregnancy rate.

Exposure to human chorionic gonadotropin during in vitro maturation does not improve the maturation rate and developmental potential of immature oocytes from patients with polycystic ovary syndrome.

OBJECTIVE: To investigate the effects of human chorionic gonadotropin (hCG) in culture medium on the in vitro maturation (IVM) and subsequent developmental potential of human immature oocytes. DESIGN: Prospective, randomized study. SETTING: Reproductive Medicine Center, First Affiliated Hospital of Wenzhou Medical College, People's Republic of China. PATIENT(S): 62 women with polycystic ovary syndrome (PCOS) undergoing IVM treatment. INTERVENTION(S): Immature oocytes were retrieved from unstimulated ovaries of women with PCOS. We tried three different culture systems in this experiment. In group A, oocytes were cultured in the medium containing M199 + 20% fetal bovine serum (FBS) + 75 mIU/mL recombinant follicle-stimulating hormone (FSH) + 0.5 IU/mL recombinant hCG. In group B, oocytes were cultured in hCG-free IVM medium (M199 + 20% FBS + 75 mIU/mL recombinant FSH) for 10 hours, then transferred to the same medium as used for group A. In group C, oocytes were cultured only in hCG-free IVM medium. After the oocytes had matured in vitro, fertilization and embryo transfer were performed. MAIN OUTCOME MEASURE(S): Rates of maturation, fertilization, cleavage, implantation, clinical pregnancy, miscarriage, and live birth. RESULT(S): For groups A, B, and C, the maturation rates at 32 and 48 hours were 46.02% and 69.25%; 43.72% and 64.51%; and 51.87% and 67.51%, respectively. Relatively satisfactory clinical results and implantation rates were obtained in all three groups. No statistically significant differences among groups A, B, and C were found in the rates of maturation, fertilization, cleavage, implantation, clinical pregnancy, miscarriage, or live birth. CONCLUSION(S): The results of our study indicated that the addition of hCG to in vitro culture medium did not improve the maturation rate or development potential of immature oocytes. For the IVM and development of immature oocytes from women with PCOS, hCG appears to be unnecessary.

[In vitro maturation and fertilization of unstimulated immature oocyte for the treatment of infertile women].

OBJECTIVE: To evaluate the efficacy of in vitro maturation (IVM) and fertilization of unstimulated immature oocytes for the treatment of infertile women. METHODS: Fifty-four cycles of IVM were carried out in 40 patients including 26 women with infertility due to polycystic ovary syndrome (PCOS), and 14 patients with history of assisted reproductive technology (ART) failure. Transvaginal ultrasound-guided oocyte collection was performed during menstrual cycle days 9 - 12 without pretreatment of gonadotropins. After 24 - 48 hours of culture, the metaphase II stage oocytes were inseminated by intracytoplasmic sperm injection (ICSI). Embryo transfer was performed 2 or 3 days after ICSI. Laser assisted hatching was done before embryo transfer. RESULTS: Seven cycles were cancelled and the cancel rate was 13% (7/54). A total of 857 immature oocytes were obtained with the mean numbers of 18.2 per cycle, and 632 oocytes developed at MII stage (73.7%, 632/857). A total of 476 oocytes were fertilized by ICSI (75.3%, 476/632), with a cleavage rate of 91.2% (434/476). Embryo transfer was performed in 47 cycles and the mean number of embryos transferred were 4.3 per cycle (range: 2 - 6). The mean endometrial thickness on the day of embryo transfer was 8.9 mm. Nineteen clinical pregnancies were obtained, giving a pregnancy rate of 35% (19/54) per start cycle and 40% (19/47) per transfer cycle. CONCLUSIONS: IVM of unstimulated immature oocytes for the treatment of women with various causes of infertility especially due to PCOS is an effective alternative method. The clinical pregnancy rate of 40% (19/47) is similar to that by conventional in vitro fertilization treatment in our unit.

[Analysis of pregnancy outcomes after in vitro fertilization-embryo transfer and intracytoplasmic sperm injection].

OBJECTIVE: To compare pregnancy and perinatal outcomes between in vitro fertilization-embryo transfer (IVF-ET) and intracytoplasmic sperm injection (ICSI). METHODS: A retrospective study was carried out to measure pre-clinical and clinical abortion, ectopic pregnancies, multiple gestations, birth weight, gestational age, congenital malformation and perinatal mortality in patients receiving either IVF-ET (n = 143, group 1) or ICSI (n = 173, group 2) from January 1999 to June 2001. The outcomes of singleton and twin were compared separately. RESULTS: The maternal age, infertility duration, parity and the number of transferred embryo were comparable between the two groups. There were no significant differences in abortion rate (16.1% vs 13.3%), birth rate (65.7% vs 74.6%) between IVF-ET and ICSI groups (P > 0.05). In singleton, the rates of low birth weight, small for gestational age and pre-term birth were 1.8%, 7.3%, 5.5% respectively in IVF-ET group and 6.8%, 8.1%, 14.9% respectively in ICSI group. In twin, the rates of low birth weight, small for gestational age and pre-term birth were 34.2%, 30.3%, 42.1% respectively in IVF-ET group and 42.6%, 38.0%, 46.3% respectively in ICSI group. There were no significant differences between the two groups (P > 0.05). But the rates of low birth weight, small for gestational age and pre-term birth were higher in twin than in singleton (P < 0.01). The incidence of congenital malformation was 2.2% and 1.6% in IVF-ET and ICSI group respectively (P > 0.05). CONCLUSIONS: The pregnancy and perinatal outcomes are similar between IVF-ET and ICSI groups. Twin is the main cause of low birth weight, small for gestational age and pre-term birth.

[Effects of metformin on gonadotropin-induced ovulation in patients with polycystic ovary syndrome].

OBJECTIVE: To evaluate the effects of metformin on gonadotropin-induced ovulation in patients with polycystic ovary syndrome (PCOS). METHODS: Forty patients with PCOS (study group) and 20 women with normal weight and menstrual cycle (control group) were enrolled. Serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), fasting glucose (FG), fasting insulin (FINS) and fasting leptin were measured before and after treatment. In the study group, 20 cases (group A) were assigned to take 500 mg of metformin three times daily for 12 weeks, if pregnancy did not occur, high purified FSH (FSH-HP) was added for one cycle; another 20 cases (group B) were induced ovulation with FSH-HP alone for one cycle. RESULTS: There were significant high FINS and leptin levels in the study group as compared with the control group [(20 +/- 16) vs (12 +/- 6) nmol/L, P < 0.05; (14 +/- 16) vs (8 +/- 4) mg/L, P < 0.05]. The obese PCOS group had markedly higher serum FINS and leptin than the non-obese PCOS group [(24 +/- 18) vs (14 +/- 8) nmol/L, P < 0.05; (20 +/- 22) vs (8 +/- 4) mg/L, P < 0.05], but serum FINS and FG were not significantly different between the non-obese PCOS and the control group (P > 0.05). After administration of metformin for 12 weeks, serum LH, T, leptin and FINS decreased significantly (P < 0.05 - 0.01), serum FSH levels and body mass index showed a slight decrease, whereas no change was found in FG. In the study group, 3 cases conceived during metformin therapy, the remaining 37 were induced ovulation with FSH-HP or FSH-HP and metformin, 7 cases obtained pregnancy. The rates of ovulation and pregnancy in group A were higher than those in group B (88% vs 70%, 24% vs 15%), but no significant difference was found. CONCLUSIONS: Metformin therapy in PCOS can decrease the FINS and leptin levels, normalize the endocrine abnormalities, resumes ovulation and pregnancy in some patients, and may improve the ovarian response to gonadotropin.

[Endocrine and metabolic effects of metformin in combination with compound cyproterone acetate in women with polycystic ovarian syndrome].

OBJECTIVE: To study the endocrinologic and metabolic effects of metformin in combination with compound cyproterone acetate (CPA) on patients with polycystic ovarian syndrome (PCOS). METHODS: A prospective study involved total 45 PCOS patients as group A and 20 non-PCOS infertility patients as control (group B). Complete baseline work-up including body mass index (BMI), waist/hip ratio (WHR), ferriman-Gallwey score (FGS), gonadotrophin, testosterone (T), sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (Ds), insulin (FI) and glucose tolerance test, were performed in all patients. Patients in group A were treated with CPA alone (group A1), metformin alone (group A2) or combination of CPA with metformin (group A3), respectively by randomization. At the end of 12-week therapy, subjects were re-evaluated and above parameters were measured. RESULTS: Women in group A had significant increases in BMI, WHR, FGS, luteinizing hormone (LH), T, FI, insulin resistance, and significantly decrease in high-density lipoprotein (HDL)-C comparing with the control group (P < 0.01). No significant difference among A1, A2 and A3 was found at baseline. LH, T, free testosterone (FT) were significant decreased from (13.9 +/- 5.9) IU/L, (2.1 +/- 0.8) nmol/L and (2.8 +/- 2.3) nmol/L respectively to (5.8 +/- 2.2) IU/L, (1.2 +/- 0.4) nmol/L and (0.8 +/- 0.5) nmol/L respectively and SHBG was significant increased from (99 +/- 42) nmol/L to (187 +/- 64) nmol/L in group A3, when compared with LH, T and FT from (13.8 +/- 7.6) IU/L, (2.2 +/- 1.1) nmol/L and (2.5 +/- 1.9) nmol/L respectively to (11.8 +/- 6.5) IU/L, (1.8 +/- 0.8) nmol/L and (1.7 +/- 1.0) nmol/L respectively and SHBG from (99 +/- 40) nmol/L to (120 +/- 51) nmol/L in group A2 (P < 0.05 approximately 0.001). HDL-C were significantly increased from (1.5 +/- 0.3) mmol/L to (1.8 +/- 0.3) mmol/L in group A3 comparing with HDL-C from (1.5 +/- 0.4) mmol/L to (1.6 +/- 0.4) mmol/L in group A1 (P < 0.001). CONCLUSIONS: The PCOS patients treated with metformin in combination with compound cyproterone acetate may be more effective in inhibiting hyperandrogen and hypersecretion of LH than metformin alone and more obvious in improving lipid profiles than CPA alone.