Identification of the 11 β-hydroxysteroid Dehydrogenase Type 1 mRNA and Protein in Human Mononuclear Leukocytes

Abstract

The enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) catalyzes the interconversion between inactive 11-ketoglucocorticoids and their active 11β-hydroxy derivatives, such as cortisol and corticosterone. We have investigated the expression of 11β-HSD1 in freshly isolated human peripheral mononuclear leukocytes (MNL). The presence of 11β-HSD1 mRNA was demonstrated in total RNA by RT-PCR using specific primers designed on the 4th and 5th exons of the human 11β-HSD1 gene. Fragments of the expected size were consistently detected on agarose gels, and sequencing showed complete identity with the corresponding sequence deposited in GenBank. The occurrence of 11β-HSD1 protein was established by Western immunoblot analysis with a specific polyclonal antibody. Enzyme oxo-reductase activity was investigated by incubating 12 samples of MNL isolated from from 8 subjects with [³ H]cortisone and formation of cortisol was established only in 4 subjects (yield range: 0.15–1.3%) after acetylation and TLC, blank subtraction and correction for losses. 18β-Glycyrrhetinic acid, an inhibitor of 11 β-HSD1, reduced cortisol production below detection limit. Dehydrogenase activity could not be demonstrated. It is suggested that, although enzyme activity of 11β-HSD1 in circulating MNL is low, it is apparently ready for enhancement after MNL migration to sites of inflammation.

11β-hydroxysteroid dehydrogenase type 1 - human mononuclear leukocytes - inflammation - oxidative stress - receptors - glucocorticoids

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1 These authors contributed equally.

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