Visfatin-induced lipid raft redox signaling platforms and dysfunction in glomerular endothelial cells.
Adipokines have been reported to contribute to glomerular injury during obesity or diabetes mellitus. However, the mechanisms mediating the actions of various adipokines on the kidney remained elusive. The present study was performed to determine whether acid sphingomyelinase (ASM)-ceramide associated lipid raft (LR) clustering is involved in local oxidative stress in glomerular endothelial cells (GECs) induced by adipokines such as visfatin and adiponectin. Using confocal microscopy, visfatin but not adiponectin was found to increase LRs clustering in the membrane of GECs in a dose and time dependent manner. Upon visfatin stimulation ASMase activity was increased, and an aggregation of ASMase product, ceramide and NADPH oxidase subunits, gp91(phox) and p47(phox) was observed in the LR clusters, forming a LR redox signaling platform. The formation of this signaling platform was blocked by prior treatment with LR disruptor filipin, ASMase inhibitor amitriptyline, ASMase siRNA, gp91(phox) siRNA and adiponectin. Corresponding to LR clustering and aggregation of NADPH subunits, superoxide (O(2)(-)) production was significantly increased (2.7 folds) upon visfatin stimulation, as measured by electron spin resonance (ESR) spectrometry. Functionally, visfatin significantly increased the permeability of GEC layer in culture and disrupted microtubular networks, which were blocked by inhibition of LR redox signaling platform formation. In conclusion, the injurious effect of visfatin, but not adiponectin on the glomerular endothelium is associated with the formation of LR redox signaling platforms via LR clustering, which produces local oxidative stress resulting in the disruption of microtubular networks in GECs and increases the glomerular permeability.
Boini
Krishna M
KM
Department of Pharmacology and Toxicology, Medical College of Virginia Campus, Virginia Commonwealth University, Richmond, VA 23298, USA.
Zhang
Chun
C
Xia
Min
M
Han
Wei-Qing
WQ
Brimson
Christopher
C
Poklis
Justin L
JL
Li
Pin-Lan
PL
D000818:Animals; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D004578:Electron Spin Resonance Spectroscopy; D005920:Glomerular Mesangium / Q000187:drug effects* / Q000378:metabolism; D050356:Lipid Metabolism; D051379:Mice; D018613:Microscopy, Confocal; D054409:Nicotinamide Phosphoribosyltransferase / Q000494:pharmacology*; D010084:Oxidation-Reduction; D015398:Signal Transduction; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D054409:Nicotinamide Phosphoribosyltransferase