Abstract
Abstract 20097: Shear-Sensitive Lipid Phosphate Phosphatase 3 Maintains Nitric Oxide-Mediated Flow-Induced Dilation In Healthy Human Arterioles
Circulation (New York, N.Y.), Vol.134(Suppl_1 Suppl 1), p.A20097
11/11/2016
DOI: 10.1161/circ.134.suppl_1.20097
Abstract
Shear-stimulated release of nitric oxide (NO) is the most important physiological endothelium-dependent vasodilator response (flow-induced dilation; FID) in the arteriolar circulation. However, in patients with coronary artery disease (CAD), NO bioavailability is reduced and the mediator of FID switches to hydrogen peroxide (H2O2), a pro-inflammatory agent. Interestingly, reduced expression of the shear-sensitive lipid phosphate phosphatase 3 (LPP3) also produces endothelial inflammation, reduction in NO and an increase in H2O2.ObjectiveWe hypothesize that reduced expression of LPP3 leads to disruption of NO-mediated FID in human adipose arterioles and elicits a switch to H2O2 as the mediator of FID. We propose that restoring expression of LPP3 in patients with CAD restores NO-mediated FID.MethodsAdipose arterioles isolated from subjects with and without CAD were prepared for videomicroscopy. Decreased expression of LPP3 in healthy vessels was achieved by LPP3-targeted siRNA. Anti-miR92a, an inhibitor of miR92a, which decreases expression of LPP3, was used to increase expression of LPP3 in vessels from CAD patients. RT-qPCR in cultured endothelial cells was used to test the effects of decreased expression of LPP3 on endothelial nitric oxide synthase (eNOS) expression.ResultsIn healthy vessels, FID was maintained but NO was no longer involved after siRNA-mediated knockdown of LPP3 (% max dilationsiRNA 74.2±5.62, n=10 vs siRNA+L-NAME [NOS inhibitor; 100μM] 84.8±4.46 n=6). Anti-miR92a restored NO-mediated FID in CAD vessels (anti-miR92a 73.0±4.29, n=6 vs anti-miR92a+L-NAME -6.30±20.6 n=3) while phenylboronic acid (PBAH2O2 scavenger; 5μM) had no effect (anti-miR92a 73.0±4.29, n=6 vs anti-miR92a+PBA 70.6±8.91, n=3). Knock-down of LPP3 in cultured endothelial cells led to 34% reduction in eNOS mRNA expression (normalized to GAPDH; n=2) in comparison to vehicle control.ConclusionReduced expression of LPP3 leads to disruption of NO-mediated FID in healthy human arterioles, recapitulating the situation in CAD. Inhibition of miR92a, which decreases LPP3 protein expression in vessels from patients with CAD restores NO-mediated dilation. Normal function/expression of LPP3 is necessary to maintain NO-mediated FID.
Details
- Title: Subtitle
- Abstract 20097: Shear-Sensitive Lipid Phosphate Phosphatase 3 Maintains Nitric Oxide-Mediated Flow-Induced Dilation In Healthy Human Arterioles
- Creators
- Dawid Chabowski - Medical College of WisconsinKarima Ait-Aissa - Medical College of WisconsinAndrew Kadlec - Medical College of WisconsinJoseph Hockenberry - Medical College of WisconsinAndreas Beyer - Medical College of WisconsinDavid Gutterman - Medical College of Wisconsin
- Resource Type
- Abstract
- Publication Details
- Circulation (New York, N.Y.), Vol.134(Suppl_1 Suppl 1), p.A20097
- Publisher
- by the American College of Cardiology Foundation and the American Heart Association, Inc
- DOI
- 10.1161/circ.134.suppl_1.20097
- ISSN
- 0009-7322
- eISSN
- 1524-4539
- Language
- English
- Date published
- 11/11/2016
- Academic Unit
- Cardiovascular Medicine; Internal Medicine
- Record Identifier
- 9984361567702771
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