Figure 5.

Effects of antioxidant treatment on PDGF-induced CSE expression, ROS production and proliferation. Quiescent mesangial cells were pre-treated with the NADPH oxidase inhibitor DPI (10 µM) or the ROS scavenger NAC (10 mM) or vehicle for 30 min. Thereafter, PDGF-BB (25 ng·mL⁻¹) was added for a further 8 h, and CSE mRNA and protein levels, as well as ROS formation, were analysed by RT-PCR and Western blotting or DCF fluorescence respectively. CSE mRNA levels are shown in panel A, protein levels in panel B. Data are means + SD (n= 3 for mRNA and protein), *P < 0.05, **P < 0.01, ***P < 0.001 significantly different from vehicle-treated controls; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 significantly different from PDGF-BB alone. The effects of PDGF-BB (8 h) as well as NAC and DPI on ROS formation measured as DCF fluorescence is shown in panel C, ***P < 0.001significantly different from vehicle treated controls; ^###P < 0.001 significantly different from PDGF-BB. (D) Quiescent rat mesangial cells were treated with or without PDG-BB (25 ng·mL⁻¹) in the presence or absence of GYY4137 (500 µM) and NAC (10 mM) for 14 h. Thereafter, 1 µCi·mL⁻¹[³H]methyl-thymidine was added, and the cells were incubated for a further 24 h. Incorporation of [³H]methyl-thymidine was assessed as described. Data are means + SD (n= 3), ***P < 0.001 significantly different from vehicle treated controls; ^##P < 0.01, ^###P < 0.001 significantly different from PDGF-BB.