Skip to main content
. 2020 Mar 25;9(4):275. doi: 10.3390/antiox9040275

Figure 4.

Figure 4

Corylin reduces VCAM-1 expression in TNF-α-treated VSMCs through ROS/MAP kinase inhibition. VSMCs were treated with 20 µM corylin, 2 mM NAC or 10 µM DPI for 1 h and then incubated with or without 10 ng/mL TNF-α for 15 min. (A) Phosphorylated P38, ERK, and JNK levels were examined by Immunoblot analysis. Total ERK (T-ERK), total P38 (T-P38), and total JNK (T-JNK) were used as an internal control for sample loading. (B) VSMCs were pretreated (1 h) with corylin (20 µM), a P38 inhibitor (SB203580; SB; 10, 20, or 30 µM), PD98059 (PD; 10, 20, or 30 µM), or a JNK inhibitor (SP600125; SP; 10, 20, or 30 nM) and then treated with 10 ng/mL TNF-α for 24 h. Western blot analysis for VCAM-1 and quantification of VCAM-1 to GAPDH in VSMCs. (C–E) After P38, ERK and JNK silencing, cells were treated with TNF-α and corylin. VCAM-1 expression was determined by Western blot analysis. Total P38 (T-P38), total ERK (T-ERK), and total JNK (T-JNK) proteins were used to examine the siRNA effects. GAPDH was used as an internal control for sample loading. (F,G) Confluent VSMCs were pretreated (1 h) with 20 µM corylin (C20), 30 µM SB203580 (SB), 30 µM PD98059 (PD), 30 nM SP600125 (SP), 10 µM parthenolide (PAR), and 1 µg/mL anti-VCAM-1 antibody or IgG and then treated with 10 ng/mL TNF-α (T) for 24 h. BCECF/AM-labeled U937 cells were added and cocultured for another 1 h. The adherent U937 cells were counted to evaluate the overall VCAM-1 expression level. The data are shown as the mean ± SD. * p < 0.05 versus the untreated group (CTRL). † p < 0.05 versus the TNF-α-treated group. Scale bars = 100 μm (F).