Influence of cytokines on peripheral metabolism of glucocorticoids

11β-hydroxysteroid dehydrogenase (11βHSD) participates in prereceptor metabolism of glucocorticoids by conversion of their active forms (cortisol or corticosterone) to inactive forms (cortisone or 11-dehydrocorticosterone), thereby regulating concentration and activity of glucocorticoids in organism. 11βHSD type 1 (11βHSD1) activates glucocorticoids from their biologically inactive keto-forms, whereas the role of 11βHSD type 2 (11βHSD2) is their biological reinactivation. Previous in vivo studies showed that mRNA expression and enzyme activity of 11βHSD1 or 2 can be modified by influence of these cytokines, for example IL-1β and TNF-α. The aim of this study was to determine whether these cytokines play a role in regulation of 11βHSD expression in distal colon organ cultures and whether the changes of 11βHSD2 expression are induced by a direct effect of cytokines on epithelial cells. Total RNA was isolated from all samples and RNA was transformed to cDNA by reverse transcription. Subsequently mRNA of 11βHSD1 or 2 and the given housekeeping gene was measured by Q-PCR. Incubation of distal colon organ cultures with TNF-α (10 ng/ml; 48 hours) caused significant increase of 11βHSD1 expression, but no influence on 11βHSD2 expression was observed. In contrast, the expression of 11βHSD2 but not 11βHSD1 was..