Impact of Metformin Treatment on Human Placental Energy Production and Oxidative Stress

Abstract

<jats:p>Metformin is increasingly prescribed in pregnancy, with beneficial maternal effects. However, it is not known how metformin-treatment impacts metabolism and energy production in the developing feto-placental unit. We assessed the human placental response to metformin using both <jats:italic>in vivo</jats:italic> and <jats:italic>in vitro</jats:italic> treated samples. trophoblasts were derived from placentas collected from non-laboured Caesarean deliveries at term, then treated <jats:italic>in vitro</jats:italic> with metformin (0.01 mM, 0.1 mM or vehicle). Metformin-concentrations were measured using liquid-chromatography mass-spectrometry. Oxygen consumption in cultured-trophoblasts was measured using a Seahorse-XF Mito Stress Test. Markers of oxidative-stress were assayed using qRT-PCR. Metformin-transporter mRNA and protein-levels were determined by quantitative RT-PCR and Western-blotting respectively. Metformin concentrations were also measured in sample trios (maternal plasma/fetal plasma/placental tissue) from pregnancies exposed to metformin on clinical-grounds. Maternal and fetal metformin concentrations <jats:italic>in vivo</jats:italic> were highly correlated over a range of concentrations (R<jats:sup>2</jats:sup> = 0.76, <jats:italic>p</jats:italic> &amp;lt; 0.001; average fetal:maternal ratio 1.5; range 0.8–2.1). Basal respiration in trophoblasts was reduced by metformin treatment (0.01 mM metformin; <jats:italic>p</jats:italic> &amp;lt; 0.05, 0.1 mM metformin; <jats:italic>p</jats:italic> &amp;lt; 0.001). Mitochondrial-dependent ATP production and proton leak were reduced after treatment with metformin (<jats:italic>p</jats:italic> &amp;lt; 0.001). Oxidative stress markers were significantly reduced in primary-trophoblast-cultures following treatment with metformin. There is a close linear relationship between placental, fetal, and maternal metformin concentrations. Primary-trophoblast cultures exposed to clinically-relevant metformin concentrations have reduced mitochondrial-respiration, mitochondrial-dependent ATP-production, and reduced markers of oxidative-stress. Given the crucial role of placental energy-production in supporting fetal growth and well-being during pregnancy, the implications of these findings are concerning for intrauterine fetal growth and longer-term metabolic programming in metformin-exposed pregnancies.</jats:p>