Metabolomic analysis of placenta tissue in obese pregnancies / C. Fattuoni, C. Mandò, F. Palmas, G.M. Anelli, C. Novielli, E. Parejo Laudicina, V.M. Savasi, L. Barberini, A. Dessì, R. Pintus, V. Fanos, A. Noto, I. Cetin. - In: JOURNAL OF PEDIATRIC AND NEONATAL INDIVIDUALIZED MEDICINE. - ISSN 2281-0692. - 6:2(2017 Oct 24), pp. 81-82. ((Intervento presentato al 13. convegno International Workshop on Neonatology tenutosi a Cagliari nel 2017.

Metabolomic analysis of placenta tissue in obese pregnancies

C. Mandò;G.M. Anelli;C. Novielli;V.M. Savasi;I. Cetin

INTRODUCTION: The placenta plays a pivotal role as it represents the connection between mother and foetus. It acts as a carrier (nutrients and oxygen, waste products and carbonic anhydride) as well as a barrier against infection. Pregnancy related metabolomics has examined different biofluids in order to reach deeper insights into pregnancy pathologies [1]. Amniotic fluid, urine and plasma were the most studied, while placenta tissue has been the subject of few studies [2]. In this report, a method to analyse the placental metabolome through the GC-MS (Gas Chromatography-Mass Spectrometry) technique is described. MATERIALS AND METHODS: Placenta tissues were collected after elective caesarean section from obese and normal weight mothers, snap frozen and stored at -80°C until analysis. Each sample was homogenized with a mixture of solvents (chloroform, methanol, water), centrifuged, and the obtained phases were separated and processed following two different protocols. The hydrophilic phase was vacuum dried and treated with two derivatizing reagents, methoxyamine hydrochloride and MSTFA (N-Methyl-N-[trimethylsilyl] trifluoroacetamide), to give a mixture of trimethylsilylated metabolites. The lipophilic phase was dried and treated with boron trifluoride in methanol to obtain a mixture of FAME (Fatty Acid Methyl Esters). The two phases were separately analysed on a GC-MS platform, applying different instrumental conditions. The chromatograms obtained were analysed with the free software AMDIS (Automated Mass Spectral Deconvolution and Identification System) using a lab-made library comprising 200 metabolites. RESULTS: The analysis of the placenta tissue allowed to identify essential metabolites. In the hydrophilic fraction, 78 metabolites were detected: they mainly belong to the classes of amino acids (18), carboxylic acids (13), sugars and sugar-related compounds (11), phosphorylated derivatives (9). The lipophilic fraction analysis allowed to define the fatty acid profile of the placenta extract: the saturated fatty acids palmitic and stearic, the monounsaturated oleic, and the polyunsaturated (PUFA) linoleic, arachidonic, 8,11,14-eicosatrienoic and 4,7,10,13,16,19-docosahexaenoic acid (DHA) were the most representative. Statistical analysis of the data matrices derived from the hydrophilic and lipophilic fraction analysis were performed in MetaboAnalyst 3.0 [3], to investigate the possibility of identifying characteristic metabolic differences between the different phenotypes. Placentae from obese mothers were compared with those from normal weight mothers to get new insights into the correlation between the metabolite content and metabolic status. CONCLUSIONS: The reported analysis method allowed to examine different metabolites in placenta samples, divided into two phases, hydrophilic and lipophilic. All metabolites were analysed through the same GC-MS platform to provide interesting informations about the placenta tissue composition. REFERENCES [1] Fanos V, Atzori L, Makarenko K, Melis GB, Ferrazzi E. Metabolomics application in maternal-fetal medicine. Biomed Res Int. 2013;2013: 720514. [2] Dunn WB, Brown M, Worton SA, Davies K, Jones RL, Kell DB, Heazell AEP. The metabolome of human placental tissue: Investigation of first trimester [3] Xia J, Sinelnikov IV, Han B, Wishart DS. MetaboAnalyst 3.0 – making metabolomics more meaningful. Nucleic Acids Res. 2015;43(W1): W251-7.
Settore MED/40 - Ginecologia e Ostetricia
Settore BIO/11 - Biologia Molecolare
Settore BIO/12 - Biochimica Clinica e Biologia Molecolare Clinica
Official Journal UENPS (Union of European Neonatal & Perinatal Societies)
Official Journal of the Portuguese Neonatal Society (affiliated to the Portuguese Paediatric Society)
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