Relationship among MAPKs, NF-κB, and Caspase-3 in VSMCs induced by SS and AGEs.

<p>The cultured quiescent VSMCs were pretreated with DMSO, NF-κB inhibitor PDTC (20 μM) and Caspase-3 inhibitor Z-DEVD-FMK (20 μM), ERK inhibitor PD98059 (50 μM), JNK inhibitor SP600125 (20 μM) and p38 inhibitor SB202190 (20 μM) for 1 h and then treated with AGEs or SS as indicated above. <b>(A)</b> Western Blot analysis showed cells pretreated with PDTC and Z-DEVD-FMK had no effect on phosphorylation of MAPKs. <b>(B)</b> Graph bars showed densitometry analysis of MAPKs activation levels normalized with total MAPKs. <b>(C)</b> Western Blot analysis showed cells pretreated with PDTC suppressed the activation of both NF-κB/p65 (Ser536), and Z-DEVD-FMK also inhibited the Caspase-3 activation while had no effect on NF-κB/p65 (Ser536) phosphorylation of VSMCs. <b>(D)</b> Graph bars showed densitometry analysis of NF-κB/p65 (Ser536) activation levels after PDTC and Z-DEVD-FMK pretreatment normalized with NF-κB. <b>(E)</b> Graph bars showed densitometry analysis of Caspase-3 activation levels after PDTC and Z-DEVD-FMK pretreatment normalized with β-actin. <b>(F)</b> Western Blot analysis showed PD98059 significantly inhibited phosphorylation of NF-κB/p65 (Ser536), while it had no influence on Caspase-3 activation. <b>(G)</b> Graph bars showed densitometry analysis of NF-κB/p65 (Ser536) activation levels after PD98059 pretreatment normalized with NF-κB. <b>(H)</b> Graph bars showed densitometry analysis of Caspase-3 activation levels after PD98059 pretreatment normalized with β-actin. <b>(I)</b> Western Blot analysis showed NF-κB/p65 (Ser536) and Caspase-3 activation could be suppressed by both SP600125 and SB202190. <b>(J)</b> Graph bars showed densitometry analysis of NF-κB/p65 (Ser536) activation levels after SP600125 and SB202190 pretreatment normalized with NF-κB. <b>(K)</b> Graph bars showed densitometry analysis of Caspase-3 activation levels after SP600125 and SB202190 pretreatment normalized with β-actin. All the experiments were independently repeated three times and shown as mean±SEM. a above bars represented the p<0.05 compared to NC group, b represented the p<0.05 compared to AGEs and SS group and c represented the p<0.05 compared to DMSO group, n = 3 (a, b and c for p value<0.05).