Figure 7.

The effects of PDI inhibitor, PDI-SiRNA and NADPH oxidase inhibitor on simultaneous increases in proliferation and apoptosis of VSMCs induced by SS and/or AGEs. The quiescent cultured VSMCs were pretreated with DMSO (w–i) (a–d) or PDI inhibitor bacitracin (e–h), or NADPH oxidase inhibitor (apocynin) (u–x) for 1 h, or transfected with SiRNA-PDI (SiR) (m–p), siRNA-control (SiC) (q–t) and Lipofectamine (Lip) (i–l). All cells above pretreated or not were subjected to SS and/or AGEs for 1 h and continually cultured for additional 24 h. These cells were stained with the primary Ki-67 antibody, Cy3-conjugated secondary antibody, and a TUNEL kit and then counterstained with DAPI. Ki-67-positive cells are shown in red, TUNEL-positive cells in green, and the nuclei of VSMCs in blue. (y, z) Statistical results of the ratio of Ki-67- or TUNEL-positive cells from a to x were obtained from three independent experiments. a, P<0.05 versus negative control (NC); b, P<0.05 versus SS or AGEs; c, P<0.05 versus without inhibitors of the same group; d, P<0.05 versus SiC of the same group by ANOVA with LSD test. Scale bar, 100 μm. Data are shown as the means±SEM