c Type Natriuretic Peptide, Hyperandrogenism, Menstrual Irregularity, Polycystic Ovary Syndrome
Conditions
Keywords
c type natriuretic peptide, polycystic ovary syndrome, menstrual cycle
Brief summary
Recent studies have shown that C natriuretic peptide is produced from granulosa cells, increasing cumulative guanosine monophosphate (cGMP) production by affecting cumulus cells through natriuretic peptide receptors. It is suggested that produced cGMP maintains the transport of oocytes via the gap junctions and leads to a continuous increase in cyclic adenosine monophosphate (cAMP) levels in the oocyte. An important role of increased internal cAMP levels in the oocyte is shown to suppress meiotic progression. Deoxyribonucleic acid studies in animals have shown that expression of the natriuretic peptide precursor increases during the periovulatory period and shows that this increase decreases rapidly after luteinizing hormone / human chorionic gonadotropin (hCG) stimulation.Human studies have shown that after ovulation induction, the CNP level in follicular fluid decreases following ovulatory dose of hCG.Polycystic ovary syndrome (PCOS) is the most common endocrine disease in the reproductive period, characterized by hyperandrogenism, oligo-anovulation, and polycystic ovarian morphology on ultrasonography, and in an animal study investigating the relationship between CNP and PCOS, serum CNP levels were increased in polycystic ovary syndrome.CNP serum level is thought to show differences between healthy women and women with polycystic ovary syndrome.
Detailed description
In this prospective study, a total of 60 patients are planed to be included. PCOS group will be consisted of 30 women and control group will include 30 healthy women with regular menstruation aged between 18-40 years old. PCOS diagnosis will be made according to Rotterdam criteria. Age and body mass index of all participants will be recorded. BMI will be calculated by dividing weight by height in square meters. Then morning fasting venous blood samples will be taken from the patients between 2nd-5th day of menstruation for both groups. All blood samples will be centrifuged on the day of collection. Sera will be aliquoted into 1.5 mL Eppendorf (Eppendorf, Milano, Italy) tubes, and will be kept at -80°C until the day of CNP test. For the PCOS patients describing oligo/anovulation, after excluding pregnancy, progesterone withdrawal bleeding will be created and then the patients will be evaluated. Serum levels of LH, FSH, estradiol, thyroid stimulating hormone (TSH), prolactin (PRL), androstenedione, dehydroepiandrosterone sulfate (DHEAS), total testosterone, free testosterone, sex hormone binding globulin (SHBG), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL), glucose and insulin levels will be analyzed. For insulin sensitivity, homeostatic model of insulin resistance (HOMA-IR) will be used and it will be calculated by the formula: HOMA-IR ¼ fasting blood glucose (mg/dL)fasting insulin (mIU/mL)/405. Free androgen index will be calculated by the formula 100x (Total testosteron/SHBG). Serum CNP levels of the patients will be analyzed by an enzyme-linked immunosorbent (ELISA) assay for human C-type natriuretic peptide in accordance with the manufacturer's instructions (SEA721Hu, ELISA Kit for Human C-Type Natriuretic Peptide, Wuhan USCN Business Co., Ltd., Cloud-Clone Corp., CCC, USA). Data will be analyzed using Statistical Packege for Social Sciences software (SPSS v15, SPSS Inc, Chicago, IL, USA). Independent t-test will be used to compare the parameters with normal distribution. Parameters that don't fulfill the parametric test assumptions will be compared using Mann-Whitney U test. Correlation of CNP with other parameters will be analyzed using Spearman's rank correlation test. Receiver operating characteristic (ROC) curve will be used to evaluate diagnostic sensitivity and specificity of CNP for PCOS. P values less than 0.05 will be regarded as statistically significant.
Interventions
DIAGNOSTIC_TESTC type natriuretic peptide
Serum level of CNP in PCOS and healthy women
Sponsors
Near East University, Turkey
Study design
Allocation
NON_RANDOMIZED
Intervention model
PARALLEL
Primary purpose
DIAGNOSTIC
Masking
NONE
Eligibility
Sex/Gender
FEMALE
Age
18 Years to 40 Years
Healthy volunteers
Yes
Inclusion criteria
* PCOS defined according to Rotterdam criteria * Healthy normal menstruating women
Exclusion criteria
* Diabetes mellitus * Cardiavascular disease * Renal disease * Any drug usage * Smoking * Pregnancy
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Serum CNP Levels of PCOS and Healthy Participants | Second or Third Day of Menstruation | The comparison of serum levels of C type natriuretic peptide among PCOS and healthy women |
| Roc Curve of CNP | Second or Third Day of Menstruation | We made a cut off value estimation for the diagnosis of PCOS by serum CNP levels using Receiver operator characteristics. The ROC curve incorporates both sensitivity (true positive rate) and specificity (true negative rate) providing a single assessment incorporating both measures. The higher the total area under the curve, the greater the predictive power of the CNP for diagnosing PCOS |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Serum Tiroid Stimulating Hormone of PCOS and Healthy Women | Second or Third Day of Menstruation | serum tiroid stimulating hormone of PCOS and healthy women were compared |
| Serum Prolactin Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | serum prolactin levels of PCOS and healthy women were compared |
| Serum Androstenedione Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum Androstenedione Levels of PCOS and Healthy Women were compared |
| Serum Dehydroepiandrosterone Sulfate (DHEAS) Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum dehydroepiandrosterone sulfate (DHEAS) Levels of PCOS and Healthy Women were compared |
| Serum Total Testosterone Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum Total Testosterone Levels of PCOS and Healthy Women were compared |
| Serum Free Testosterone Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum Free Testosterone Levels of PCOS and Healthy Women were compared |
| Comparison of FSH and LH of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum FSH and LH of PCOS and healthy women were compared |
| Free Androgen Index of Participants | Second or Third Day of Menstruation | Free androgen index of participants were compared. It was calculated with the formula 100XTotal testosterone/Sex hormone binding globulin |
| Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | Second or third day of menstruation and morning fasting | Serum glucose, total cholesterol and triglycerides levels of participants were compared |
| High Density Lipoprotein and Low Density Lipoprotein Levels of Participants | Second or Third Day of Menstruation-morning fasting | High density lipoprotein and low density lipoprotein levels of participants were compared |
| Serum Insulin Levels of Participants | Second or Third Day of Menstruation-morning fasting | Serum Insulin Levels of Participants were compared |
| Homeostatic Model Assessment of Insulin Resistance of Participants | Second or Third Day of Menstruation-morning fasting | homeostatic model assessment of insulin resistance of participants were compared. The calculation was made with formula Glucose X Insulin/405. |
| Serum Sex Hormone Binding Globulin Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum Sex Hormone Binding Globulin Levels of PCOS and Healthy Women were compared |
| Serum Estradiol Levels of PCOS and Healthy Women | Second or Third Day of Menstruation | Serum Estradiol levels of PCOS and Healthy Women were compared |
Countries
Cyprus
Participant flow
Recruitment details
The patients were recruited from Near East University Hospital, Department of Obstetrics and Gynecology.
Pre-assignment details
For the last 6 months, patients with drug use that may affect menstruation like oral contraceptives, patients with cardiac or renal disease and therefore drug use, patients with hypertension hystory, history of ovarian surgery, current pregnancy, hyperprolactinemia, thyroid disease, congenital adrenal hyperplasia.
Participants by arm
| Arm | Count |
|---|---|
| Women With Polycystic Ovary Syndrome 36 patients with PCOS
C type natriuretic peptide: Serum level of CNP in PCOS and healthy women | 36 |
| Healthy Women of Reproductive Age 30 patients with regular normal menstrual cycle
C type natriuretic peptide: Serum level of CNP in PCOS and healthy women | 30 |
| Total | 66 |
Withdrawals & dropouts
| Period | Reason | FG000 | FG001 |
|---|---|---|---|
| Overall Study | insufficient sample volume | 2 | 3 |
| Overall Study | missing serum samples | 2 | 2 |
| Overall Study | Withdrawal by Subject | 8 | 7 |
Baseline characteristics
| Characteristic | Women With Polycystic Ovary Syndrome | Healthy Women of Reproductive Age | Total |
|---|---|---|---|
| Age, Categorical <=18 years | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical >=65 years | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical Between 18 and 65 years | 36 Participants | 30 Participants | 66 Participants |
| Age, Continuous | 22 years | 24 years | 23 years |
| Body Mass Index | 23.20 kg/m^2 STANDARD_DEVIATION 4.26 | 21.17 kg/m^2 STANDARD_DEVIATION 1.88 | 22.28 kg/m^2 STANDARD_DEVIATION 3.52 |
| Race/Ethnicity, Customized Cypriot | 36 Participants | 30 Participants | 66 Participants |
| Region of Enrollment Cyprus | 36 participants | 30 participants | 66 participants |
| Sex: Female, Male Female | 36 Participants | 30 Participants | 66 Participants |
| Sex: Female, Male Male | 0 Participants | 0 Participants | 0 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk |
|---|---|---|
| deaths Total, all-cause mortality | 0 / 0 | 0 / 0 |
| other Total, other adverse events | 0 / 0 | 0 / 0 |
| serious Total, serious adverse events | 0 / 0 | 0 / 0 |
Outcome results
Primary
Roc Curve of CNP
We made a cut off value estimation for the diagnosis of PCOS by serum CNP levels using Receiver operator characteristics. The ROC curve incorporates both sensitivity (true positive rate) and specificity (true negative rate) providing a single assessment incorporating both measures. The higher the total area under the curve, the greater the predictive power of the CNP for diagnosing PCOS
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (NUMBER) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Roc Curve of CNP | 0.706 probability |
p-value: 0.004Roc curve
Primary
Serum CNP Levels of PCOS and Healthy Participants
The comparison of serum levels of C type natriuretic peptide among PCOS and healthy women
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum CNP Levels of PCOS and Healthy Participants | 110 pg/ml |
| Healthy Women of Reproductive Age | Serum CNP Levels of PCOS and Healthy Participants | 57 pg/ml |
p-value: 0.004Wilcoxon (Mann-Whitney)
Secondary
Comparison of FSH and LH of PCOS and Healthy Women
Serum FSH and LH of PCOS and healthy women were compared
Time frame: Second or Third Day of Menstruation
Population: FSH and LH should be evaluated during menstruation time period ( day 3)
| Arm | Measure | Group | Value (MEDIAN) |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Comparison of FSH and LH of PCOS and Healthy Women | Follicle Stimulating Hormone | 4.7 mIU/mL |
| Women With Polycystic Ovary Syndrome | Comparison of FSH and LH of PCOS and Healthy Women | Luteinizing Hormone | 4.23 mIU/mL |
| Healthy Women of Reproductive Age | Comparison of FSH and LH of PCOS and Healthy Women | Follicle Stimulating Hormone | 4.97 mIU/mL |
| Healthy Women of Reproductive Age | Comparison of FSH and LH of PCOS and Healthy Women | Luteinizing Hormone | 5.36 mIU/mL |
p-value: 0.171Wilcoxon (Mann-Whitney)
p-value: 0.176Wilcoxon (Mann-Whitney)
Secondary
Free Androgen Index of Participants
Free androgen index of participants were compared. It was calculated with the formula 100XTotal testosterone/Sex hormone binding globulin
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Free Androgen Index of Participants | 4.39 % of T testosterone per SHBG | Standard Deviation 2.58 |
| Healthy Women of Reproductive Age | Free Androgen Index of Participants | 1.55 % of T testosterone per SHBG | Standard Deviation 0.89 |
p-value: <0.001t-test, 2 sided
Secondary
High Density Lipoprotein and Low Density Lipoprotein Levels of Participants
High density lipoprotein and low density lipoprotein levels of participants were compared
Time frame: Second or Third Day of Menstruation-morning fasting
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Women With Polycystic Ovary Syndrome | High Density Lipoprotein and Low Density Lipoprotein Levels of Participants | High density lipoprotein | 53.82 mg/dL | Standard Deviation 11.06 |
| Women With Polycystic Ovary Syndrome | High Density Lipoprotein and Low Density Lipoprotein Levels of Participants | Low density lipoprotein | 98.50 mg/dL | Standard Deviation 23.61 |
| Healthy Women of Reproductive Age | High Density Lipoprotein and Low Density Lipoprotein Levels of Participants | High density lipoprotein | 62.15 mg/dL | Standard Deviation 19.48 |
| Healthy Women of Reproductive Age | High Density Lipoprotein and Low Density Lipoprotein Levels of Participants | Low density lipoprotein | 92.92 mg/dL | Standard Deviation 23.56 |
p-value: 0.114t-test, 2 sided
p-value: 0.504Wilcoxon (Mann-Whitney)
Secondary
Homeostatic Model Assessment of Insulin Resistance of Participants
homeostatic model assessment of insulin resistance of participants were compared. The calculation was made with formula Glucose X Insulin/405.
Time frame: Second or Third Day of Menstruation-morning fasting
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Homeostatic Model Assessment of Insulin Resistance of Participants | 1.50 HOMA-IR Index |
| Healthy Women of Reproductive Age | Homeostatic Model Assessment of Insulin Resistance of Participants | 1.45 HOMA-IR Index |
p-value: 0.886Wilcoxon (Mann-Whitney)
Secondary
Serum Androstenedione Levels of PCOS and Healthy Women
Serum Androstenedione Levels of PCOS and Healthy Women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Androstenedione Levels of PCOS and Healthy Women | 148.89 ng/dL | Standard Deviation 52.38 |
| Healthy Women of Reproductive Age | Serum Androstenedione Levels of PCOS and Healthy Women | 84.66 ng/dL | Standard Deviation 11.56 |
p-value: <0.001t-test, 2 sided
Secondary
Serum Dehydroepiandrosterone Sulfate (DHEAS) Levels of PCOS and Healthy Women
Serum dehydroepiandrosterone sulfate (DHEAS) Levels of PCOS and Healthy Women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Dehydroepiandrosterone Sulfate (DHEAS) Levels of PCOS and Healthy Women | 304.55 µg/dL |
| Healthy Women of Reproductive Age | Serum Dehydroepiandrosterone Sulfate (DHEAS) Levels of PCOS and Healthy Women | 299.95 µg/dL |
p-value: 0.277Wilcoxon (Mann-Whitney)
Secondary
Serum Estradiol Levels of PCOS and Healthy Women
Serum Estradiol levels of PCOS and Healthy Women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Estradiol Levels of PCOS and Healthy Women | 31 pg/mL |
| Healthy Women of Reproductive Age | Serum Estradiol Levels of PCOS and Healthy Women | 36.5 pg/mL |
p-value: 0.306Wilcoxon (Mann-Whitney)
Secondary
Serum Free Testosterone Levels of PCOS and Healthy Women
Serum Free Testosterone Levels of PCOS and Healthy Women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Free Testosterone Levels of PCOS and Healthy Women | 1.71 pg/mL | Standard Deviation 0.64 |
| Healthy Women of Reproductive Age | Serum Free Testosterone Levels of PCOS and Healthy Women | 1.12 pg/mL | Standard Deviation 0.3 |
p-value: <0.001t-test, 2 sided
Secondary
Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants
Serum glucose, total cholesterol and triglycerides levels of participants were compared
Time frame: Second or third day of menstruation and morning fasting
| Arm | Measure | Group | Value (MEDIAN) |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | Total cholesterol | 164 mg/dL |
| Women With Polycystic Ovary Syndrome | Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | triglycerides | 78 mg/dL |
| Women With Polycystic Ovary Syndrome | Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | Glucose | 88.5 mg/dL |
| Healthy Women of Reproductive Age | Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | Total cholesterol | 171 mg/dL |
| Healthy Women of Reproductive Age | Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | triglycerides | 77 mg/dL |
| Healthy Women of Reproductive Age | Serum Glucose, Total Cholesterol and Triglycerides Levels of Participants | Glucose | 89 mg/dL |
p-value: 0.947Wilcoxon (Mann-Whitney)
p-value: 0.906Wilcoxon (Mann-Whitney)
p-value: 0.428Wilcoxon (Mann-Whitney)
Secondary
Serum Insulin Levels of Participants
Serum Insulin Levels of Participants were compared
Time frame: Second or Third Day of Menstruation-morning fasting
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Insulin Levels of Participants | 7.10 mU/L |
| Healthy Women of Reproductive Age | Serum Insulin Levels of Participants | 7.10 mU/L |
p-value: 0.713Wilcoxon (Mann-Whitney)
Secondary
Serum Prolactin Levels of PCOS and Healthy Women
serum prolactin levels of PCOS and healthy women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Prolactin Levels of PCOS and Healthy Women | 17.21 ng/mL | Standard Deviation 6.65 |
| Healthy Women of Reproductive Age | Serum Prolactin Levels of PCOS and Healthy Women | 16.80 ng/mL | Standard Deviation 6.26 |
p-value: 0.852t-test, 2 sided
Secondary
Serum Sex Hormone Binding Globulin Levels of PCOS and Healthy Women
Serum Sex Hormone Binding Globulin Levels of PCOS and Healthy Women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Sex Hormone Binding Globulin Levels of PCOS and Healthy Women | 30.4 nmol/L |
| Healthy Women of Reproductive Age | Serum Sex Hormone Binding Globulin Levels of PCOS and Healthy Women | 59.4 nmol/L |
p-value: 0.014Wilcoxon (Mann-Whitney)
Secondary
Serum Tiroid Stimulating Hormone of PCOS and Healthy Women
serum tiroid stimulating hormone of PCOS and healthy women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEDIAN) |
|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Tiroid Stimulating Hormone of PCOS and Healthy Women | 1.61 mIU/L |
| Healthy Women of Reproductive Age | Serum Tiroid Stimulating Hormone of PCOS and Healthy Women | 1.47 mIU/L |
p-value: 0.795Wilcoxon (Mann-Whitney)
Secondary
Serum Total Testosterone Levels of PCOS and Healthy Women
Serum Total Testosterone Levels of PCOS and Healthy Women were compared
Time frame: Second or Third Day of Menstruation
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Women With Polycystic Ovary Syndrome | Serum Total Testosterone Levels of PCOS and Healthy Women | 1.38 nmol/L | Standard Deviation 0.48 |
| Healthy Women of Reproductive Age | Serum Total Testosterone Levels of PCOS and Healthy Women | 0.92 nmol/L | Standard Deviation 0.36 |
p-value: 0.006t-test, 2 sided