Journal
FREE RADICAL RESEARCH
Volume 42, Issue 9, Pages 789-797Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/10715760802400766
Keywords
Vascular calcification; uremia; osteoblastic differentiation; NAD(P)H oxidase; -glycerophosphate; oxidative stress
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Funding
- Amgen Inc
- Societe Francophone de Dialyse
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Recent studies showed that hydrogen peroxide (H2O2) enhanced bone markers expression in vascular smooth muscle cells (VSMCs) implicated in osteoblastic differentiation. This study aimed at investigating the role of NAD(P)H oxidase in vascular calcification processes. A7r5 rat VSMCs were incubated with -glycerophosphate (10 mm) or uremic serum to induce a diffuse mineralization. H2O2 production by VSMCs was determinated by chemiluminescence. NAD(P)H oxidase sub-unit (p22phox), Cbfa-1, ERK phosphorylation and bone alkaline phosphatase (ALP) expressions were measured by Western blotting. VSMCs exhibited higher production of H2O2 and early expression of p22phox with -glycerophosphate or uremic serum within 24 h of treatment. -glycerophosphate-induced oxidative stress was associated with Cbfa-1 expression followed by ALP expression and activity, meanwhile the VSMCs expressing ALP diffusely calcified their extracellular matrix. Interestingly, diphenyleneiodonium partly prevented the osteoblastic differentiation. Results from this model strongly suggest a major implication of vascular NAD(P)H oxidase in vascular calcification supported by VSMCs osteoblastic differentiation.
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