Introduction NAFLD is characterised by steatosis, chronic inflammation and fibrosis. The underlying mechanisms include insulin resistance, increased free fatty acid flux from de-novo lipogenesis and decreased lipid oxidation. Vascular Adhesion Protein—1 (VAP -1), is an adhesion molecule with semicarbazide- sensitive amine oxidase activity (SSAO), which is also expressed as a soluble protein in serum (sVAP-1) and elevated in inflammatory liver diseases such as NAFLD. VAP-1 has been shown to modulate glucose and lipid uptake in muscle and adipose tissue and thus we investigated whether it may contribute to glucose and lipid homeostasis in human liver tissue.
Methods We have used precision cut liver slices (PCLS) from normal and diseased human liver specimens and cultured human sinusoidal endothelium (HSEC) and hepatocyte cells in combination with VAP-1 substrates (200 μM methylamine or benzylamine) and inhibitors (400 μM bromoethylamine) to perform standard ex vivo radiolabelled glucose uptake and fatty acid uptake assays using oil red O quantification following exposure of cells to Oleic and Palmitic acid (PA). Immunohistochemical staining and qPCR were performed using standard techniques and for confirmatory experiments HSEC were transfected with enzymatically active/inactive VAP1.
Results QPCR confirmed upregulation of VAP-1 mRNA (DDCT=1.144 p=0.03) in NASH vs normal liver and also changes in FABP1, -4, -5, FATP3, -4 (p≤0.05 for all) and GLUT-1, 2, 3, 5, 8, 9 and 12 in NAFLD compared to normal individuals. Results were confirmed using immunohistochemical staining. Exposure of human PCLS to sVAP-1 and methylamine typically resulted in a 38%–54% increase in PA uptake (p≤0.01 for all) and a 20% increase in hepatocyte glucose uptake in vitro which could be inhibited using bromoethylamine. Transduction of HSEC with enzymatically active VAP-1 also increased glucose uptake which was prevented in the absence of enzyme activity. Interestingly methylamine treatment of human liver resulted in decreased expression of mRNA for glucose transporters and an increase in some lipid transporters including FABP6, FATP and LRP8, and H2O2 produced by SSAO activity increase lipid uptake by hepatic cells.
Conclusion In conclusion, we demonstrate for the first time global alterations in cellular expression of glucose and lipid transporter proteins in human NAFLD. We confirm that VAP-1 is elevated in disease and that SSAO activity of VAP-1 results in enhanced hepatic lipid and glucose uptake and changes in transporter expression. Thus we propose that bioactive metabolites of SSAO activity contribute to the metabolic derangement evident in fatty liver disease.
Competing interests None declared.
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.