METHODS: CYP11B2 was inhibited in HAC15 cells using siRNA (siCYP11B2) or baxdrostat, a selective CYP11B2 inhibitor. Forty-eight hours post-treatment, steroid hormone levels, RNA sequencing profiles, and functional outputs (EZMTT metabolic assay, xCELLigence real-time cell analysis, and Annexin V/SYTOX flow cytometry for apoptosis/necrosis) were evaluated.
RESULTS: siCYP11B2 reduced aldosterone production by 70% ± 2.9% (n = 18, p n = 9, p n = 12, p n = 12, p n = 144 and p = 1.3 × 10-10); however, the relative distribution of apoptotic and necrotic populations varied across independent experiments.
CONCLUSION: Reduced aldosterone synthesis is associated with transcriptional signatures involving autophagy- and mitophagy-related pathways. Pharmacological treatment using U0126 and SB203580 altered steroidogenic output independently of CYP11B2 transcript expression, providing hypothesis-generating evidence linking cellular homeostasis and mitochondrial quality control processes to adrenal steroidogenesis. Furthermore, pharmacological CYP11B2 inhibition using baxdrostat produced variable alterations in cell-death profile, highlighting important considerations for long-term therapeutic targeting.