Histone Deacetylase Inhibitor Impairs Plasminogen Activator Inhibitor-1 Expression via Inhibiting TNF-α-Activated MAPK/AP-1 Signaling Cascade

Figure 1

Effect of CBHA on HDAC activity and PAI-1 expression in human pleural mesothelial cells. (a) MeT-5A cells were treated with TNF-α (10 ng/mL) for the indicated times or pretreated with CBHA (1 μM) for 15 min before stimulation with TNF-α for 2 hours. Total HDAC enzyme activity was determined by using the HDAC fluorometric cellular activity assay. (b) MeT-5A cells and (c) primary cultured human pleural mesothelial cells were pretreated with CBHA, respectively, followed by stimulation with TNF-α (10 ng/mL) for 24 h. The levels of PAI-1 were assessed by Western blot. (d) MeT-5A cells were pretreated with CBHA (0.5–2 μM), followed by stimulation with TNF-α (10 ng/mL) for 6 h. PAI-1 mRNA concentrations were analyzed by semiquantitative reverse transcriptase PCR and normalized with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. Band intensity was quantified as described in Section 2. Data are shown as mean ± SEM of three independent experiments. ^##𝑃<0.01 and ^###𝑃<0.001 compared with the resting group; *𝑃<0.05, **𝑃<0.01, and ***𝑃<0.001 compared with the vehicle (DMSO) group; ^†𝑃<0.05 compared with the 2 hr TNF-α-treated group.

(a)

(b)

(c)

(d)