Lipopolysaccharide and double stranded viral RNA mediate insulin resistance and increase system a amino acid transport in human trophoblast cells in vitro.

INTRODUCTION: Inflammation and underlying low-grade maternal infection can impair insulin signalling and upregulate nutrient transport in the placenta which contribute to fetal overgrowth associated with GDM and/or obese pregnancies. There are, however, no studies on the role of infection on placental nutrient transport in pregnancies complicated by GDM and/or obesity. Thus, the aims of this study were to determine the effect of the bacterial product lipopolysaccharide (LPS) or the viral dsRNA analogue polyinosinic:polycytidylic acid (poly(I:C)) on the insulin signalling pathway and amino acid transport in primary human trophoblast cells.

METHODS: Human primary villous trophoblast cells were treated with LPS or poly(I:C). Protein expression of insulin signalling pathway proteins, insulin receptor (IR)-β, insulin receptor substrate (IRS)-1 and protein kinase B (also known as Akt), and phosphatidylinositol-4,5-bisphosphate 3-kinase p85α subunit (PI3K-p85α) protein were assessed by Western blotting. Glucose and amino acid uptake were assessed by radiolabelled assay. Western blotting and qRT-PCR were used to determine amino acid transporter protein and mRNA expression, respectively.

RESULTS: LPS and poly(I:C) significantly decreased phosphorylation of IR-β, IRS-1, Akt, total PI3K-p85α protein expression and glucose uptake. LPS and poly(I:C) also significantly increased expression of System A amino acid transporters SNAT1 and SNAT2, and System A-mediated uptake of amino acids.

DISCUSSION: LPS and poly(I:C) induces insulin resistance and increases amino acid uptake in human primary trophoblast cells. This suggests that the presence of low-grade maternal infection can contribute to excess placental nutrient availability and promote fetal overgrowth in pregnancies complicated by GDM and/or obesity.