Detection of total reactive oxygen species (ROS) in human aortic endothelial cells (HAECs).

(A) L5 (50 µg/mL) from patients increased ROS production (green) in HAECs after 20 minutes (top and middle row) and after 24 hours (bottom row). Compared to the phosphate-buffered saline (PBS) control, L1 treatment had no effect. Staining with 4′,6-diamidino-2-phenylindole (DAPI) (blue) indicated HAEC apoptosis 24 hours after L5 (50 µg/mL) exposure. The pretreatment of cells with TS92 attenuated the L5-induced ROS increase after 20 minutes (middle row). (B) Blocking the production of ROS by adding ROS inhibitor N-acetyl cysteine (NAC) attenuated the L5-induced increase in CRP levels. For all experiments, n = 3, and bars shown in graphs represent standard deviation. *P<0.05, **P<0.01 vs. untreated control.