Carbon Monoxide-Releasing Molecule-3 Suppresses Tumor Necrosis Factor-α- and Interleukin-1β-Induced Expression of Junctional Molecules on Human Gingival Fibroblasts via the Heme Oxygenase-1 Pathway

Figure 5

Effects of CORM-3 on HO-1 mRNA and protein expression in HGFs incubated with TNF-α and IL-1β. (a) HGFs were divided into six groups: control; TNF-α+IL-1β, in which HGFs were incubated with 10 ng/ml TNF-α and 2 ng/ml IL-1β for 24 h; CORM-3, in which HGFs were treated with 200 μM CORM-3 for 24 h; CORM-3+TNF-α+IL-1β, in which HGFs were treated with 200 μM CORM-3 and 10 ng/ml TNF-α and 2 ng/ml IL-1β for 24 h; iCORM-3, in which HGFs were treated with 200 μM inactivated CORM-3 for 24 h; and iCORM-3+TNF-α+IL-1β, in which cells were incubated with 200 μM inactivated CORM-3 and 10 ng/ml TNF-α and 2 ng/ml IL-1β for 24 h. HO-1 mRNA expression levels were determined by RT-qPCR. (b) Protein expression levels of HO-1 were determined by western blotting. β-Actin was used to demonstrate equal sample loading. The values were obtained from at least three independent experiments performed in triplicate, where , , and versus TNF-α- and IL-1β-induced cells and ^{^}, ^{^^}, and ^{^^^} compared between two groups, according to ANOVA followed by Tukey’s multiple comparison test. HO-1: heme oxygenase-1; CORM-3: carbon monoxide-releasing molecule-3; TNF-α: tumor necrosis factor-α; IL-1β: interleukin-1β; HGFs: human gingival fibroblasts.

(a)

(b)