Membrane raft-lysosome redox signalling platforms in coronary endothelial dysfunction induced by adipokine visfatin.
OBJECTIVE
The adipokine visfatin, produced during obesity, has been reported to participate in the development of cardiovascular disease associated with metabolic syndrome. The present study was designed to test a hypothesis that visfatin causes coronary endothelial dysfunction through lysosome trafficking and fusion to cell membranes, membrane raft (MR) clustering, and formation of redox signalosomes.


RESULTS
By using confocal microscopy, it was found that visfatin, but not adiponectin, stimulated NADPH oxidase (NOX) subunits, gp91(phox) aggregation in MR clusters and p47(phox) translocation to these MR clusters in bovine coronary arterial endothelial cells (CAECs), leading to activation of NOX with a 2.5-fold increase in O(2)(·-) production. A signalling lipid, ceramide, was found to be enriched in such membrane MR-NOX complexes of CAECs. Lysosomal fluorescent dye (FM1-43) quenching and de-quenching revealed that visfatin induced the fusion of lysosomes to cell membranes and incorporation of acid sphingomyelinase and its product, ceramide, in such MR-NOX signalling platforms. Functionally, visfatin significantly attenuated endothelium-dependent vasodilation in small coronary arteries (by 80%), which was blocked by lysosomal function inhibitor and MR disruptors.


CONCLUSIONS
These results suggest that lysosome-associated molecular trafficking and consequent ceramide accumulation in cell membrane may mediate the assembly of NOX subunits and their activation in response to adipokine visfatin in CAECs, thereby producing endothelial dysfunction in coronary circulation.
Xia
Min
M
Department of Pharmacology and Toxicology, Medical College of Virginia Campus, Virginia Commonwealth University, 410 N 12th Street, Richmond, VA 23298, USA.
Zhang
Chun
C
Boini
Krishna M
KM
Thacker
Audrey M
AM
Li
Pin-Lan
PL
D052242:Adiponectin / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002518:Ceramides / Q000378:metabolism; D003331:Coronary Vessels / Q000187:drug effects / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology*; D008561:Membrane Fusion; D021962:Membrane Microdomains / Q000187:drug effects / Q000201:enzymology*; D018613:Microscopy, Confocal; D019255:NADPH Oxidases / Q000378:metabolism; D054409:Nicotinamide Phosphoribosyltransferase / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D021381:Protein Transport; D011994:Recombinant Proteins; D015398:Signal Transduction / Q000187:drug effects; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors; D014664:Vasodilation; D014665:Vasodilator Agents / Q000494:pharmacology
D052242:Adiponectin; D002518:Ceramides; D011994:Recombinant Proteins; D014665:Vasodilator Agents; D013481:Superoxides; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D054409:Nicotinamide Phosphoribosyltransferase; D013108:Sphingomyelin Phosphodiesterase
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