223 Investigating the role of aerobic glycolysis in arterial calcification

Objective The process of arterial calcification shares many similarities to skeletal mineralisation, and involves the deposition of hydroxyapatite in the arteries. However, the cellular mechanisms responsible have yet to be fully elucidated. Accumulating evidence suggests that aerobic glycolysis (the Warburg effect), plays a critical role in meeting the demand for energy and biosynthetic precursors during proliferation and differentiation in numerous cell types. Therefore we addressed the hypothesis that vascular smooth muscle cell (VSMC) calcification requires aerobic glycolysis to produce energy and the necessary biosynthetic precursors. Methods Calcification of murine aortic VSMCs was induced by 3 mM Pi for 7 days. Calcium deposition was determined using alizarin red staining and a modified o-cresolphthalein method. VSMCs were cultured with the fluorescent glucose analogue 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)−2-Deoxyglucose (2-NBDG) to determine changes in glucose uptake. Gene expression was analysed by qRT-PCR. Results Calcium deposition was significantly increased in VSMCs cultured in 3 mM Pi versus control conditions (124%, p Conclusion Together these data suggest that arterial calcification requires glucose metabolism through a mechanism involving Wnt signalling. Interruption of the glycolysis pathway may therefore represent a novel therapeutic target for clinical intervention.