期刊
KIDNEY INTERNATIONAL
卷 86, 期 1, 页码 58-66出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/ki.2013.520
关键词
endothelial cell; endothelial nitric oxide synthase; Fabry disease; globotriaosylceramide
资金
- National Institutes of Health [5R01DK055823-13, DK89503, DK097153]
The endothelial dysfunction of Fabry disease results from alpha-galactosidase A deficiency leading to the accumulation of globotriaosylceramide. Vasculopathy in the alpha-galactosidase A null mouse is manifested as oxidant-induced thrombosis, accelerated atherogenesis, and impaired arterial reactivity. To better understand the pathogenesis of Fabry disease in humans, we generated a human cell model by using RNA interference. Hybrid endothelial cells were transiently transfected with small interfering RNA (siRNA) specifically directed against alpha-galactosidase A. Knockdown of alpha-galactosidase A was confirmed using immunoblotting and globotriaosylceramide accumulation. Endothelial nitric oxide synthase (eNOS) activity was correspondingly decreased by >60%. Levels of 3-nitrotyrosine (3NT), a specific marker for reactive nitrogen species and quantified using mass spectrometry, increased by 40- to 120-fold without corresponding changes in other oxidized amino acids, consistent with eNOS-derived reactive nitrogen species as the source of the reactive oxygen species. eNOS uncoupling was confirmed by the observed increase in free plasma and protein-bound aortic 3NT levels in the alpha-galactosidase A knockout mice. Finally, 3NT levels, assayed in biobanked plasma samples from patients with classical Fabry disease, were over sixfold elevated compared with age- and gender-matched controls. Thus, 3NT may serve as a biomarker for the vascular involvement in Fabry disease.
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