Hydroxytyrosol Ameliorates Endothelial Function under Inflammatory Conditions by Preventing Mitochondrial Dysfunction

Figure 5

HT improves mitochondrial function in PMA-stimulated endothelial cells. HUVEC were pretreated with HT (1–30 μmol/L) for 1 h and then stimulated with PMA (10 nmol/L) for further 16 h. After treatments, MMP was assayed by using JC-1 staining and evaluated by using a fluorescence plate reader (a). Mitochondrial oligomycin-sensitive ATP synthesis was measured in endothelial cells incubated in the absence (−) or in the presence of HT (1–30 μmol/L) (b). The expression of β subunit of ATP synthase was evaluated at protein (c) and mRNA levels (d) by Western blotting or quantitative RT-PCR, respectively. ATP5β protein expression was normalized to β-actin, and ATP5β mRNA amount was normalized to Gapdh mRNA. Data are representative of four independent experiments (mean ± SD), each consisting of four replicates for each condition, and expressed as percentage of PMA-stimulated endothelial cells. versus CTR; versus PMA alone.

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