Delphinidin-3-Glucoside Protects against Oxidized Low-Density Lipoprotein-Induced Mitochondrial Dysfunction in Vascular Endothelial Cells via the Sodium-Dependent Glucose Transporter SGLT1
Figure 5
Determination of Dp uptake by VECs.
(A) The autofluorescent emission spectra of Dp and ethanol (baseline). The peak emission wavelength was recorded at approximately 370 nm. (B) HUVECs were treated with 10 μM Dp for 2 h and the subcellular Dp distribution was measured using a CLSM (b, ×400 and d, ×1000 magnification) and with a contrast phase microscope (a, ×400 and c,×1000 magnification). (C) Evaluation of intracellular Dp content in HUVECs via HPLC. After incubation, cells were lysed and the supernatant was collected for HPLC analysis. The blank control cells was treated with 0.1% DMSO. The excitation spectra of the fluorescence for λem = 370 nm is presented in arbitrary units (a.u.). (D-E) Time, dos-, and temperature-dependent uptake of Dp in HUVECs was measured via HPLC. (D) HUVECs were treated with 10 μM Dp at 37 (▪) and 4°C (⧫) for the indicated times (10 min, 30 min, 1 h, 2 h, 4 h, and 24 h, respectively). (E) HUVECs were treated with different Dp concentrations (1, 10, 20, 50, and 100 μM, respectively) for 2 h at 37°C (▪) or 4°C (⧫), respectively. The difference between incorporated Dp by HUVECs at 37°C and that at 4°C represents the actively transported amount (▴). BABL/c mice were intravenously administrated different Dp doses or an equivalent volume of hydro-alcoholic solution. (F) Representative image showed the isolated thoracic aorta (red arrow). The Dp content in the blood (G) and isolated thoracic aorta (H) were measured by HPLC. The isolated thoracic aortas were separated, frozen sectioned, and then stained with PI to visualize the nuclei. (I) Representative fluorescent signals (white arrows) of Dp in blood vessels were observed by by CLSM (TCS SP2, Leica Microsystems GmbH). Values are presented as means ± SEM, n = 3.