Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB223

Poster Communications

Role of glycolysis in regulation of endothelial barrier integrity in inflammation

M. Aslam1, C. Hamm1, D. Guenduez1

1. Kerckhoff Herzforschungsinstitut, Justus Liebig University Giessen, Bad Nauheim, Hessen, Germany.

Background: The integrity of the endothelial cell (EC) barrier of the microvasculature is compromised in inflammation and increased vascular permeability is a surrogate marker for the development of atherosclerotic lesions. In recent years, considerable advances have been made in the understanding of signalling mechanisms regulating the endothelial barrier integrity. The role of endothelial metabolism as a modulator of endothelial barrier integrity is not yet well-studied particularly in the setting of lung vascular injury. The aim of the present study was to investigate the role of glycolysis regulatory enzymes PFK1 and PFK-FB3 in the maintenance of endothelial barrier integrity. Methods: The study was carried out on cultured human umbilical vein endothelial cells (ECs) and rat coronary microvascular ECs. Endothelial barrier function was analysed by measuring the flux of albumen through EC monolayers cultured on filter membranes. Gene expression was analysed by qPCR-based assays. Results: Under inflammatory conditions (TNFa; 1 ng/mL for 24h), an upregulation of glycolysis regulatory enzymes PFK1 and PFK-FB3 was observed in endothelial cells (n=5; p<0.05) at mRNA level. This was accompanied by an increase in total cellular ATP levels. Pharmacological inhibition or genetic depletion of either PFK1 or PFK-FB3 resulted in reduction of cellular ATP content and worsened TNFa-induced endothelial hyperpermeability. On the other hand, over expression of PFK-FB3 ameliorated TNFa-induced endothelial hyperpermeability. These data suggest that upregulation of glycolysis may serve as an endogenous metabolic adaptation to inflammatory vascular injury and protect endothelial barrier integrity.

Where applicable, experiments conform with Society ethical requirements