Mechanisms of angiotensin II-mediated activation of aldosterone synthase in H295R human adrenocortical and rat adrenal glomerulosa cells

Mária Szekeres , Gábor Turu , Anna Orient , Bence Szalai , Katinka Süpeki , Miklós Cserző , Péter Várnai , László Hunyady

In adrenal zona glomerulosa cells angiotensin II (Ang II) is a key regulator of steroidogenesis. Our purpose was to compare the mechanisms of Ang II-induced changes in the expression level of early transcription factors NR4A1 (NGFIB) and NR4A2 (Nurr1) genes, and the CYP11B2 gene encoding aldosterone synthase in H295R human adrenocortical tumor cells and in primary rat adrenal glomerulosa cells. Real-time PCR studies have demonstrated that Ang II increased the expression levels of NR4A1 and NR4A2 in H295R cells within one hour after stimulation, which persisted up to 6 hrs; whereas in rat adrenal glomerulosa cells the kinetics of the expression of these genes were more rapid and transient. Ang II also induced prolonged nuclear translocation of Nurr1 and NGFIB proteins in both cell types. Studies using MEK inhibitor (PD98059, 20μM), protein kinase C inhibitor (BIM1, 3μM) and calmodulin kinase (CAMK) inhibitor (KN93, 10μM) revealed that in rat adrenal glomerulosa cells CAMK-mediated mechanisms play a predominant role in the regulation of CYP11B2. In accordance with earlier findings, in H295R cells MEK inhibition increased the expression of NR4A1, NR4A2 and CYP11B2 genes, however, it decreased the Ang II-induced gene expression levels, suggesting that ERK activation has a role in control of expression of these genes. No such mechanism was detected in rat glomerulosa cells. Sar-Ile-Ile-AngII, which can cause G protein-independent ERK activation, also stimulated the expression of CYP11B2 in H295R cells. These data suggest that the previously reported CAMK-mediated stimulation of early transcription factors NGFIB and Nurr1 has a predominant role in Ang II-induced CYP11B2 activation in rat adrenal glomerulosa cells, whereas in H295R cells ERK activation and G protein-independent mechanisms also contribute to this process.

Mots clés: Adrenal cortex; Aldosterone synthase; Angiotensin II; Calmodulin kinase; Gene expression

Mária Szekeres, Gábor Turu, Anna Orient, Bence Szalai, Katinka Süpeki, et al.. Mechanisms of angiotensin II-mediated activation of aldosterone synthase in H295R human adrenocortical and rat adrenal glomerulosa cells. Molecular and Cellular Endocrinology, 2009, 302 (2), pp.244. ⟨10.1016/j.mce.2008.12.015⟩. ⟨hal-00499108⟩