Gas chromatography-mass spectrometry-based metabolic profiling of androgens, progestins and glucocorticoids in women with polycystic ovary syndrome

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Abstract


Hypothesis/aims of study. Polycystic ovary syndrome (PCOS) is a common disease. Depending on the diagnostic criteria, the disease is seen in 10-20% of women of reproductive age and accounts for 70-80% of all forms of hyperandrogenic syndrome. PCOS is a heterogeneous condition of multifactorial etiology characterized by various clinical, endocrine and metabolic disorders. Therefore, it is important to clarify the specific features of steroid hormone biosynthesis and metabolism and steroidogenesis enzyme activity, as well as to search for new laboratory criteria for early diagnosis and prompt treatment. The aim of this study was to perform metabolic profiling of androgens, progestins and glucocorticoids using gas chromatography-mass spectrometry (GC-MS) in obese and non-obese women with PCOS.

Study design, materials and methods. We examined 53 women of reproductive age diagnosed with PCOS. The first group included 30 women aged 22 to 29 years with normal body weight. The second group comprised 23 obese patients aged 25 to 33 years with an average body mass index (BMI) of 35.3 ± 0.4 kg/m2. The control group consisted of 25 healthy women aged 26 ± 0.6 years having a normal BMI without clinical and biochemical signs of hyperandrogenism. Immunoassay methods were used to determine the serum levels of luteinizing hormone, follicle-stimulating hormone, free testosterone, 17-hydroxyprogesterone, and sex hormone-binding globulin. A glucose tolerance test was performed to determine glucose and insulin levels before and after load. Urine steroid profiles were studied by GC-MS with the optimization of the sample preparation schedule. Statistical data processing was performed using the STATISTICA for WINDOWS software system (version 10). The main quantitative characteristics of the patients are presented as the median (Me), the 25th percentile and the 75th percentile (Q25Q75). To compare the results obtained in the study groups, the nonparametric Mann-Whitney test was used. The 95% confidence interval was considered statistically significant.

Results. The article presents a metabolomics analysis of androgens, glucocorticoid hormones and progestins in women with PCOS compared to the control group. It was revealed that non-obese patients with PCOS had increased urinary excretion of androstenedione metabolites, dehydroepiandrosterone and its metabolites, 17-hydroxypregnanolone, pregnantriol, and 5-ene-pregnenes, while obese patients with PCOS had increased that of androsterone and dehydroepiandrosterone metabolites (16-oxo-androstenediol and androstenediol-17β) compared to the control group findings. Decreased ratios of cortisol and cortisone tetrahydro metabolite amount to the levels of 11-oxo-pregnanetriol, pregnanetriol and 17-hydroxypregnenolone, when compared to the control group, was obtained in non-obese patients with PCOS, which indicates 21-hydroxylase deficiency. In obese patients with PCOS, four signs of increased 5α-reductase activity were obtained, and in PCOS patients with a normal BMI, three signs were obtained, which indicates varying 5α-reductase activity in PCOS patients depending on the BMI.

Conclusion. Quantitative evaluation of androgen and progestin metabolites, as well as 5α- and 5β-metabolites of androstenedione and glucocorticoids in the study of urine steroid profiles by GC-MS method opens new opportunities for PCOS diagnostics.


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About the authors

Natalya V. Vorokhobina

North-Western State Medical University named after I.I. Mechnikov

Author for correspondence.
Email: natalya.vorokhobina@szgmu.ru
ORCID iD: 0000-0002-9574-105X
SPIN-code: 4062-6409

Russian Federation, Saint Petersburg

MD, PhD, DSci (Medicine), Professor, Head of the Department of Endocrinology named after academician V.G. Baranov

Lyudmila I. Velikanova

North-Western State Medical University named after I.I. Mechnikov

Email: velikanova46@gmail.com
ORCID iD: 0000-0002-9352-4035

Russian Federation, Saint Petersburg

PhD, DSci (Biology), Professor, Head of the Research Laboratory of Chromatography

Olga B. Glavnova

The Research Institute of Obstetrics, Gynecology, and Reproductology named after D.O. Ott

Email: o.glavnova@mail.ru
ORCID iD: 0000-0001-6087-252X

Russian Federation, Saint Petersburg

MD

Ekaterina V. Malevanaya

North-Western State Medical University named after I.I. Mechnikov

Email: obedkovaev@gmail.com
ORCID iD: 0000-0003-0880-0814

Russian Federation, Saint Petersburg

PhD, Senior Researcher

Ravilya K. Galakhova

North-Western State Medical University named after I.I. Mechnikov

Email: rgalakhova@gmail.com
ORCID iD: 0000-0003-3599-3199
SPIN-code: 1865-2310

Russian Federation, Saint Petersburg

MD, PhD, Associate Professor

Irina Yu. Matezius

North-Western State Medical University named after I.I. Mechnikov

Email: imatezius@rambler.ru
ORCID iD: 0000-0002-8694-9756
SPIN-code: 7421-1610

Russian Federation, Saint Petersburg

MD, PhD, Associate Professor. The Department of Endocrinology named after academician V.G. Baranov

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Supplementary files

Supplementary Files Action
1.
Fig. 1. Urinary excretion of androgens in normal weight and obese patients with polycystic ovary syndrome (data obtained using gas chromatography-mass spectrometry). PCOS, polycystic ovary syndrome; BMI, body mass index; An, androsterone; Et, etiocholanolone; DHEA, dehydroepiandrosterone; dA2, androstenediol; dA3, androstenetriol

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2.
Fig. 2. Urinary excretion of glucocorticoid tetrahydro metabolites in normal weight and obese patients with polycystic ovary syndrome (data obtained using gas chromatography-mass spectrometry). PCOS, polycystic ovary syndrome; BMI, body mass index; THS, tetrahydro-11-deoxycortisol; THE, tetrahydrocortisone; ТНВ, tetrahydrocorticosterone

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3.
Fig. 3. Urinary excretion of 17-hydroxyprogesterone metabolites and 5-ene-pregnenes in normal weight and obese patients with polycystic ovary syndrome (data obtained using gas chromatography-mass spectrometry). PCOS, polycystic ovary syndrome; BMI, body mass index; 17P, 17-hydroxypregnenolone; P3, pregnanetriol; P2, pregnanediol; dP2, pregnenediol; dP3, pregnenetriol

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Copyright (c) 2020 Vorokhobina N.V., Velikanova L.I., Glavnova O.B., Malevanaya E.V., Galakhova R.K., Matezius I.Y.

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