Fig. 2

The effect of HDAC3 inhibition in T2DM–induced endothelial dysfunction in vitro. a Fluorescent images of superoxide levels in HUVECs cultured either in Con (5.5 mM of glucose), MAN (33 mM: 5.5 mM of glucose + 27.5 mM of D-mannitol) or HG (33 mM)-PA (100 μM) medium with or without si-HDAC3 for 72 h, MAN was served as the osmotic control for the HG-PA. Superoxide was determined with the fluorescent indicator DHE, and the fluorescent intensity of DHE was observed with a computer-assisted microscope. Scale bars: 100 μm. b Representative images of aortic rings from db/m mice were cultured in medium. Scale bars: 200 μm. c TUNEL assay of HUVECs was determined. The apoptotic cells were labeled with green, and nuclei were stained with DAPI (blue). Scale bars: 100 μm. d Capillary-like tube formation was assessed by Matrigel angiogenesis assay in HUVECs. Scale bars: 300 μm. e–h Quantification of the Fluorescence intensity in (a), number of sprouts in (b), TUNEL + cells in (c), the tube length in (d). i Cell lysates of HUVECs were used to detect Bax, Bcl-2 as well as c-Caspase 3 protein levels by immunoblotting. j, k The quantitative analysis of each immunoblotting. The data are represented as the means ± SEM (n = 5). Significance: *P < 0.05 vs. Con or MAN in scrambled HUVECs. #P < 0.05 vs. HG-PA in scrambled HUVECs. All above results in graphs from western blot were normalized to the first group