Calcineurin mediates the angiotensin II-induced aldosterone synthesis in the adrenal glands by up-regulation of transcription of the CYP11B2 gene.

Aldosterone synthesis in the zona glomerulosa of the adrenal gland is catalysed by aldosterone synthase (CYP11B2). The CYP11B2 expression is induced by angiotensin II (Ang II), mediated by increase of intracellular Ca(2+) level. Since calcineurin (CN) is an important mediator activated by Ca(2+), we investigated the issue of whether CN is involved in the Ang II-induced CYP11B2 expression in human adrenocortical H295R cells. First, CN inhibitors, cyclosporine A (CysA) and tacrolimus (FK506) inhibited the Ang II-induced elevation of CYP11B2 mRNA level. Second, enforced expression of a constitutively active CN increased the CYP11B2 mRNA level. Third, depletion of CN by siRNA technique blocked the Ang II-induced elevation of CYP11B2 mRNA level. Fourth, in reporter assays using a luciferase gene connected to a 5'-flanking region (from -134 to +43 bp) of the hCYP11B2 gene, both CysA and FK506 inhibited the Ang II-mediated up-regulation of luciferase activity. Finally, activation of CN in living H295R cells following the Ang II treatment was confirmed using a fluorescence resonance energy transfer-based sensor. Taken together, we conclude that CN mediates the Ang II-induced aldosterone synthesis through up-regulation of the CYP11B2 transcription.