Abstract W P388: High Fat and High Glucose Synergistically Impair Brain Microvascular Endothelial Cell Survival and Angiogenic Potential After Hypoxia

Diet-induced metabolic disease is the most rapidly increasing risk factor for ischemic stroke and the leading cause of long-term disability in the United States. We have shown that high fat diet-induced diabetes impedes vascular restoration and functional recovery after ischemic stroke. Accordingly, the objective of this study was to test the hypothesis that high fat and high glucose combination impairs brain microvascular endothelial cell survival and angiogenic potential after hypoxia in a synergistic manner. Primary brain microvascular endothelial cells (BMVECs) were cultured in normal glucose (Control), high glucose (HG, 12.5 mM, levels observed in the high fat diet-fed animals), palmitate (Pal, 200 μM, a saturated free fatty acid that has been shown to most rapidly increase in plasma after a high fat meal), or HG plus Pal for 14 hours before exposing to normoxia (N) or oxygen glucose deprivation (OGD), n=4-6. Cell survival (MTT assay and cell counts), angiogenic properties (migration and tube formation) and inflammatory mediators (TNFα and toll like receptor-TLR4) were measured. HG plus Pal combination significantly reduced cell number, % viability, % migration and tube length and increased TNFα and TLR4 levels especially under OGD conditions (Table). These results suggest that even mild elevations in glucose and free fatty acids promote inflammation and impair endothelial cell recovery mechanisms, and a second hit such as hypoxia exacerbates this effect.