Abstract
Aims/hypothesis
Fetuin-A (alpha2-Heremans-Schmid glycoprotein), a liver-derived circulating glycoprotein, contributes to lipid disorders, diabetes and cardiovascular diseases. In a previous study we found that perivascular fat cells (PVFCs) have a higher angiogenic potential than other fat cell types. The aim was to examine whether fetuin-A influences PVFC and vascular cell growth and the expression and secretion of proinflammatory and angiogenic proteins, and whether TLR4-independent pathways are involved.
Methods
Mono- and co-cultures of human PVFCs and endothelial cells were treated with fetuin-A and/or palmitate for 6–72 h. Proteins were quantified by ELISA and Luminex, mRNA expression by real-time PCR, and cell growth by BrDU-ELISA. Some PVFCs were preincubated with a nuclear factor κB NFκBp65 inhibitor, or the toll-like receptor 4 (TLR4) inhibitor CLI-095, or phosphoinositide 3-kinase (PI3K)/Akt inhibitors and/or stimulated with insulin. Intracellular forkhead box protein O1 (FoxO1), NFκBp65 and inhibitor of κB kinase β (IKKβ) localisation was visualised by immunostaining.
Results
PVFCs expressed and secreted IL-6, IL-8, plasminogen activator inhibitor 1 (PAI-1), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF)-BB, monocyte chemotactic protein-1 (MCP-1), vascular endothelial growth factor (VEGF), placental growth factor (PLGF) and hepatocyte growth factor (HGF). Fetuin-A upregulated IL-6 and IL-8, and this was potentiated by palmitate and blocked by CLI-095. Immunostaining and electrophoretic mobility shift assay (EMSA) showed partial NFκBp65 activation. MCP-1 was upregulated and blocked by CLI-095, but not by palmitate. However, HGF was downregulated, which was slightly potentiated by palmitate. This effect persisted after TLR4 pathway blockade. Stimulation of insulin–PI3K–Akt signalling by insulin resulted in nuclear FoxO1 extrusion and HGF upregulation. Fetuin-A counteracted these insulin effects.
Conclusions/interpretation
Fetuin-A and/or palmitate influence the expression of proinflammatory and angiogenic proteins only partially via TLR4 signalling. HGF downregulation seems to be mediated by interference with the insulin-dependent receptor tyrosine kinase pathway. Fetuin-A may also influence angiogenic and proinflammatory proteins involved in atherosclerosis.
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Abbreviations
- bFGF:
-
Basic FGF
- EC:
-
Endothelial cell
- EMSA:
-
Electrophoretic mobility shift assays
- FGF:
-
Fibroblast growth factor
- FoxO1:
-
Forkhead box protein O1
- HGF:
-
Hepatocyte growth factor
- IKKβ:
-
Inhibitor of κB kinase β
- LPS:
-
Lipopolysaccharide
- MCP-1:
-
Monocyte chemotactic protein-1
- NFκB:
-
Nuclear factor κB
- PAI-1:
-
Plasminogen activator inhibitor-1
- PDGF:
-
Platelet-derived growth factor
- PI3K:
-
Phosphoinositide 3-kinase
- PVAT:
-
Perivascular adipose tissue
- PVFC:
-
Perivascular fat cell
- PLGF:
-
Placental growth factor
- SMC:
-
Smooth muscle cell
- TLR4:
-
Toll-like receptor 4
- TSP-1:
-
Thrombospondin-1
- VEGF:
-
Vascular endothelial growth factor
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Funding
The present study was funded by the Federal Ministry for Education and Research (FkZ 01GI 0925), a grant from the German Federal Ministry of Education and Research to the German Center for Diabetes Research and a grant of from the German Federal Ministry for Education and Research (FkZ 01KQ0902F), joint research project ‘Gesundheitsregion REGINA’.
Duality of interest
The authors declare that there is no duality of interest associated with this manuscript.
Contribution statement
DIS-A substantially contributed to conception and design, acquisition of data, and analysis and interpretation of data, drafted and wrote the article, finally approved the version to be published and is responsible for the integrity of the work as a whole. FG, CW and SU contributed substantially to conception and design, data analysis and interpretation of data and revised the article critically for important intellectual content. NS, KR, US, BS and ER contributed to data acquisition, data analysis, interpretation of data and critically revised the manuscript. CK contributed to data acquisition and critically revised the manuscript. H-ES, UAS and AK contributed to data acquisition and critically revised the manuscript. H-UH designed the study and critically revised the manuscript. All authors approved the final version of the manuscript to be published.
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Siegel-Axel, D.I., Ullrich, S., Stefan, N. et al. Fetuin-A influences vascular cell growth and production of proinflammatory and angiogenic proteins by human perivascular fat cells. Diabetologia 57, 1057–1066 (2014). https://doi.org/10.1007/s00125-014-3177-0
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DOI: https://doi.org/10.1007/s00125-014-3177-0