Figure 6.

The effects of inhibitor, overexpression, SiRNA of PDI and NOX inhibitor on ROS production in VSMCs induced by SS and/or AGEs. The quiescent cultured VSMCs were treated with SS and/or AGEs for 10 min after pretreatment with DMSO (NC) (a–d), PDI inhibitor (bacitracin, PDI-I) (e–h), and NADPH oxidase inhibitor (apocynin, NOX-I) (i–l) for 1 h. The quiescent VSMCs with stable PDI overexpression or VSMCs transfected with siRNA-control (SiR-C) (u–x) or siRNA-PDI (SiR-PDI) (y, z, a’, b’) were treated by SS and/or AGEs for 10 min. All treated cells above were stained with the reactive oxygen species (ROS) probe (green) and then counterstained with Hochest 33342 (blue). ROS-positive cells are shown in green and the nuclei of VSMCs in blue. (c’) Statistical results of ratios of ROS-positive cells were obtained from (a–z, a’, b’) of three independent experiments. a, P<0.05 versus negative control (NC); b, P<0.05 versus without inhibitors of the same group; c, P<0.05 versus cells without PDI overexpression (EV); d, P<0.05 versus SiC by ANOVA with LSD test. Data are shown as the means±SEM