Figure 1.

Budesonide inhibits release of cytokines and MMPs by human lung fibroblasts.

Notes: HFL-1 cells were cultured until sub-confluence and treated with varying concentrations of budesonide for 30 minutes followed by stimulation with cytokines (IL-1β plus TNF-α, 1 ng/ml each). After 24 hours, medium was harvested for quantification of IL-6 (A) and IL-8 (B) by ELISA, and MMP-1 (C) and MMP-3 (D) by immunoblotting. (A and B): Effect on IL-6 and IL-8 release. Cell number was counted with a Coulter counter and the cytokine level was normalized to the cell number. Vertical axis: amount of IL-6 or IL-8 expressed as percentage of response in cytokines-only treated (IL-1β plus TNF-α) cells; horizontal axis: treatment with cytokines and varying concentrations of budesonide. Data presented are the mean ± SEM of three separate experiments. (C and D): Inset is an example of immunoblot image. In order to make quantitative comparisons, the same volume of media from each condition was harvested and precipitated for loading. Bar graphs presented are densitometric analysis of immunoblots expressed as the mean ± SEM of three separate experiments. Vertical axis: density of the image expressed as percentage of density in the cytokines-only treated group; horizontal axis: treatment with cytokines and varying concentrations of budesonide. *P < 0.05; **P < 0.01 compared to cytokines alone by one way analysis of variance followed by Tukey’s test.

Abbreviations: ELISA, enzyme linked immunosorbent assay; IL, interleukin; MMP, matrix metalloproteinase; SEM, standard error of the mean; TNF, tumor necrosis factor.