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Exp Clin Endocrinol Diabetes 2009; 117(9): 514-518
DOI: 10.1055/s-0028-1105924
Short Communication

© J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York

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

C. Fiore 1 [*] , A. Nardi 2 [*] , L. Dalla Valle 2 , D. Pellati 1 , Z. Krozowski 3 , L. Colombo 2 , D. Armanini 1
  • 1Department of Medical and Surgical Sciences, Endocrinology- University of Padua
  • 2Department of Biology, University of Padua
  • 3Baker Heart Research Institute, Melbourne, Australia
Further Information

Publication History

received 21.01.2008 first decision 11.04.2008

accepted 15.11.2008

Publication Date:
20 February 2009 (online)

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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.

Key words

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

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1 These authors contributed equally.

Correspondence

D. ArmaniniMD 

Department of Medical and Surgical Sciences-Endocrinology

University of Padua

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35128 Padua

Italy

Phone: +39/049/821 30 23

Fax: +39/049/65 73 91

Email: decio.armanini@unipd.it

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