Poster Presentation Annual Meetings of the Endocrine Society of Australia and Society for Reproductive Biology and Australia and New Zealand Bone and Mineral Society 2016

Mass spectrometric identification of eicosanoid and endocannabinoid production by human amnion tissue following stimulation with IL-1β and anandamide (#456)

Hassendrini N Peiris 1 , Kanchan Vaswani 1 , Yong Koh 1 , Fatema Almughlliq 1 , Leon Oh 1 , Murray D Mitchell 1
  1. The University of Queensland Centre for Clinical Research, Herston, QLD, Australia

Introduction: An increase in intrauterine prostaglandin production is critical for the onset and progression of labour in women and indeed all mammalian species studied. Endocannabinoids can act as substrates for enzymes of the prostaglandin biosynthetic pathways and can be utilized to generate other related compounds such as prostamides and prostanoids. The related end products are indistinguishable by radioimmunoassay.

Aim: To use mass spectrometry to identify products of endocannabinoid and eicosanoid biosynthetic pathways produced by amnion upon exposure to inflammatory stimuli (interleukin 1beta; IL-1β) with and without the addition of substrate (Anandamide; AEA.)

Methods: Human amnion explants from term placentae (delivery by elective caesarean section due to cephalopelvic disproportion) were treated with IL-1β (0.2 ng/mL), AEA (10µM) or a combination of IL-1β and AEA. The concentrations of eicosanoids (PGE2-EA, and PGE2) generated by amnion explants (in explant culture media) were measured by LC-MS/MS in positive and negative modes. 

Results: Amnion explants produced significantly more prostaglandins (PGE2) when treated with an inflammatory agent (IL-1β). The addition of anandamide substrate significantly increased the production of PGE2-EA (p<0.05) compared to the inflammatory agent alone. In the absence of anandamide substrate prostaglandins (PGE2) represent >90% of PGHS-derivatives. However, in the presence of anandamide substrate prostamides PGE2-EA represent >90% of PGHS-derivatives.

Conclusion: We provide evidence that there is differential regulation of prostaglandin and prostamide biosynthesis in human in amnion in response to inflammatory stimuli and substrate (AEA). Our data demonstrate that amnion responds by a differential drive through the prostaglandin biosynthetic pathways. Moreover, and importantly, this has been shown using the “gold standard” of measurement by mass spectrometric means. The possibility is raised that separation of these products might reduce variability in results and lead to potential uses for their measurement in the diagnosis of preterm labour.