title: Citrate treatment reduces endothelial death and inflammation under hyperglycaemic conditions.
abstract: Hyperglycaemia and glucose degradation products (GDPs) are closely associated with oxidative stress and inflammation in diabetic patients, a condition that leads to endothelial dysfunction and cardiovascular problems. We evaluated the effect of citrate and gluconate on glucose- and GDP-induced endothelial inflammation by measuring changes in viability, inflammation and function in primary human umbilical vein endothelial cells (HUVECs). The extent of apoptosis/necrosis was measured by flow cytometry and visualised with confocal microscopy by staining with annexin V or propidium iodide, respectively. Protein kinase C-βII (PKC-βII) activation was evaluated with Western blotting. Incubation with glucose (30 mM) and GDP (50 µM) significantly increased PKC-βII expression, endothelial cell death and inflammation. The addition of citrate decreased hyperglycaemia-induced apoptosis (p = 0.021), necrosis (p = 0.04) and reduced PKC-βII expression (p = 0.021) down to background levels. Citrate improved endothelial function by reducing the inflammatory markers (p = 0.01) and by decreasing neutrophil diapedesis (p = 0.012). These results suggest that citrate may have therapeutic potential by reducing hyperglycaemia-induced endothelial inflammation and abolishing endothelial dysfunction.
authors: Department of Nephrology, Lund University, Lund, Sweden.
authors: Anna
authors: A
authors: Bryland
authors: Anders
authors: A
authors: Wieslander
authors: Ola
authors: O
authors: Carlsson
authors: Thomas
authors: T
authors: Hellmark
authors: Gabriela
authors: G
authors: Godaly
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019343:Citric Acid / Q000494:pharmacology*; D018920:Coculture Techniques; D004789:Enzyme Activation; D005434:Flow Cytometry; D005942:Gluconates / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006943:Hyperglycemia / Q000378:metabolism* / Q000473:pathology; D007249:Inflammation / Q000378:metabolism* / Q000473:pathology; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D018613:Microscopy, Confocal; D009336:Necrosis; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D064546:Protein Kinase C beta; D011753:Pyrones / Q000378:metabolism; D057705:Transendothelial and Transepithelial Migration / Q000187:drug effects
chemical_list: C504711:3,4-dideoxyglucosone-3-ene; D000893:Anti-Inflammatory Agents; D005942:Gluconates; D011753:Pyrones; D018799:Intercellular Adhesion Molecule-1; D019343:Citric Acid; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D005947:Glucose; C030691:gluconic acid