The unconventional role of acid sphingomyelinase in regulation of retinal microangiopathy in diabetic human and animal models

Diabetes
Madalina OpreanuJulia V Busik

Abstract

Acid sphingomyelinase (ASM) is an important early responder in inflammatory cytokine signaling. The role of ASM in retinal vascular inflammation and vessel loss associated with diabetic retinopathy is not known and represents the goal of this study. Protein and gene expression profiles were determined by quantitative RT-PCR and Western blot. ASM activity was determined using Amplex Red sphingomyelinase assay. Caveolar lipid composition was analyzed by nano-electrospray ionization tandem mass spectrometry. Streptozotocin-induced diabetes and retinal ischemia-reperfusion models were used in in vivo studies. We identify endothelial caveolae-associated ASM as an essential component in mediating inflammation and vascular pathology in in vivo and in vitro models of diabetic retinopathy. Human retinal endothelial cells (HREC), in contrast with glial and epithelial cells, express the plasma membrane form of ASM that overlaps with caveolin-1. Treatment of HREC with docosahexaenoic acid (DHA) specifically reduces expression of the caveolae-associated ASM, prevents a tumor necrosis factor-α-induced increase in the ceramide-to-sphingomyelin ratio in the caveolae, and inhibits cytokine-induced inflammatory signaling. ASM is expressed in bot...Continue Reading

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Methods Mentioned

BETA
Assay
confocal microscopy
PCR

Related Concepts

Acid sphingomyelinase-1
Blood Capillaries
Vascular Permeability
Streptozotocin Diabetes
Diabetic Retinopathy
Docosahexaenoic Acid, Sodium Salt
Pecten Oculi
Sphingomyelinase
Rats, Holtzman
MRNA Differential Display

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