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 7

Expression changes of β-catenin, VE-cadherin, and ZO-1 in HGFs transfected with HO-1 siRNA. (a) HGFs were transiently transfected with HO-1 siRNA or scrambled siRNA. HGFs without transfection were used as control. mRNA expression of tight and adherens junctions was detected by RT-qPCR. (b) HGFs were divided into seven 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+TNF-α+IL-1β, in which HGFs were treated with 200 μM CORM-3, 10 ng/ml TNF-α, and 2 ng/ml IL-1β for 24 h; scrambled siRNA+TNF-α+IL-1β, in which cells were transiently transfected with scrambled siRNA 6 h prior to incubation with 10 ng/ml TNF-α and 2 ng/ml IL-1β for 24 h; scrambled siRNA+TNF-α+IL-1β+CORM-3, in which cells transiently transfected with scrambled siRNA for 6 h were incubated with 10 ng/ml TNF-α, 2 ng/ml IL-1β, and 200 μM CORM-3 for 24 h; HO-1 siRNA+TNF-α+IL-1β, in which cells were transiently transfected with HO-1 siRNA 6 h prior to incubation with 10 ng/ml TNF-α and 2 ng/ml IL-1β for 24 h; HO-1 siRNA+TNF-α+IL-1β+CORM-3, in which cells were transiently transfected with HO-1 siRNA 6 h prior to incubation with 10 ng/ml TNF-α, 2 ng/ml IL-1β, and 200 μM CORM-3 for 24 h. HO-1 mRNA expression levels were determined by RT-qPCR. (c) 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; siRNA: small interfering RNA; TNF-α: tumor necrosis factor-α; IL-1β: interleukin-1β; HGFs: human gingival fibroblasts.

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