ALDH2 Activity Reduces Mitochondrial Oxygen Reserve Capacity in Endothelial Cells and Induces Senescence Properties

Figure 2

ALDH2 silencing or inhibition impairs endothelial functions. (a) BrdU incorporation in ECs transfected with siRNA for 24 h. Proliferative capacity was assessed after 48 h treatment with 0.1%–2% FBS. Data are reported as mean ± SD. vs. siCTR with 0.1% FBS; and vs. siCTR. (b) Cell survival in 10 μM daidzin-treated ECs or siCTR and siALDH2 ECs exposed to 2% FBS for 2 and (c) 5 days. Data are expressed as means ± SD of the cell number counted/well. Dimethyl sulfoxide (DMSO) is used as a solvent to dissolve daidzin. No significant effect of DMSO was observed in HUVEC survival (DMSO-treated cell numbers/well: 329 ± 30). NS: not statistically significant; vs. Ctr; vs. siCTR. D10: 10 μM daidzin. (d) Scratch assay in 10 μM daidzin-treated ECs or in siCTR and siALDH2 ECs cultured in 0.1% or 2% FBS for 18 h. Means ± SD of % of scratch closure ( vs. Ctr; vs. siCTR). (e) Confocal analysis of VE-cadherin and ZO-1 patterns in control (i–iii) and siALDH2 ECs (ii–iv) after exposure to EBM-2 with 0.1% FBS for 8 h. Representative images of three experiments at 63x magnification are shown. Scale bar: 20 μm. (f) Permeability in siCTR and siALDH2 ECs detected as fluorescence-conjugated FITC-dextran diffusion through the confluent monolayers after exposure to EBM-2 with 0.1% FBS for 8 h. vs. siCTR. Images are representative of results obtained with siALDH2 B.

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