The HUVEC cell line was purchased from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in low-glucose Dulbecco's modified Eagle's medium (DMEM; Hyclone, GE Healthcare Life Sciences, Logan, UT, USA), supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA) and maintained in a humidified 5% CO2 atmosphere. Cells between passages 4 and 6, in the logarithmic phase of growth, were used in the subsequent experiments. The morphology of the cells was observed using a phase-contrast microscope (cellSens standard software version 1.6, DP72; Olympus Corporation, Tokyo, Japan).
Hydrogen peroxide (H2O2) is commonly used in models of oxidative stress-induced apoptosis (18 (link)). In the present study, HUVECs were treated with various concentrations of H2O2 (200, 500, 700 and 1,000 µmol/l;Table I ) using 0.3% H2O2 stock solution, or 0 µl H2O2 (low-gluocse DMEM alone containing 10% FBS) for 16 h to induce oxidative stress, as previously described (19 (link)–21 (link)).
Hydrogen peroxide (H2O2) is commonly used in models of oxidative stress-induced apoptosis (18 (link)). In the present study, HUVECs were treated with various concentrations of H2O2 (200, 500, 700 and 1,000 µmol/l;