Fig. 4.
Effect of hydroxytyrosol (HT) and polyphenol extract from extra virgin olive oil (PEEVOO) on the changes in nitric oxide (NO) and intracellular calcium concentration ([Ca2+]i) induced by acetylcholine (Ach) in the presence of high glucose (HG) and/or oleic and linoleic acids in ECV304 cells. Cells were cultured in 199 medium and 10% FBS for 48 h in normal glucose (NG, 5 mM), HG (30 mM) or HG together with linoleic or oleic acid (60 µM) in the presence or absence of HT (10 µM) or PEEVOO (10 µM gallic acid equivalents). Ach (0.1 µM) was added and NO concentration (A) and [Ca2+]i (B) were determined. Results are means±SEM of at least three experiments performed in triplicate. *p<0.05 vs control cells; ‡p<0.05 vs HG condition; †p<0.05 vs HG/linoleic acid; #p<0.05 vs HG/oleic acid.