Interruption of TRPC6-NFATC1 signaling inhibits NADPH oxidase 4 and VSMCs phenotypic switch in intracranial aneurysm

Biomed Pharmacother. 2023 May:161:114480. doi: 10.1016/j.biopha.2023.114480. Epub 2023 Mar 10.

Abstract

Intracranial aneurysm (IA) is a frequent cerebrovascular disorder with unclear pathogenesis. The vascular smooth muscle cells (VSMCs) phenotypic switch is essential for IA formation. It has been reported that Ca2+ overload and excessive reactive oxygen species (ROS) are involved in VSMCs phenotypic switch. The transient receptor potential canonical 6 (TRPC6) and NADPH oxidase 4 (NOX4) are the main pathway to participate in Ca2+ overload and ROS production in VSMCs. Ca2+ overload can activate calcineurin (CN), leading to nuclear factor of activated T cell (NFAT) dephosphorylation to regulate the target gene's transcription. We hypothesized that activation of TRPC6-NFATC1 signaling may upregulate NOX4 and involve in VSMCs phenotypic switch contributing to the progression of IA. Our results showed that the expressions of NOX4, p22phox, p47phox, TRPC6, CN and NFATC1 were significantly increased, and VSMCs underwent a significant phenotypic switch in IA tissue and cellular specimens. The VIVIT (NFATC1 inhibitor) and BI-749327 (TRPC6 inhibitor) treatment reduced the expressions of NOX4, p22phox and p47phox and the production of ROS, and significantly improved VSMCs phenotypic switch in IA rats and cells. Consistent results were obtained from IA Trpc6 knockout (Trpc6-/-) mice. Furthermore, the results also revealed that NFATC1 could regulate NOX4 transcription by binding to its promoter. Our findings reveal that interrupting the TRPC6-NFATC1 signaling inhibits NOX4 and improves VSMCs phenotypic switch in IA, and regulating Ca2+ homeostasis may be an important therapeutic strategy for IA.

Keywords: Intracranial aneurysm; NADPH oxidase 4; Nuclear factor of activated T cells; Transient receptor potential canonical 6; Vascular smooth muscle cells.

MeSH terms

  • Animals
  • Intracranial Aneurysm* / metabolism
  • Mice
  • Muscle, Smooth, Vascular / metabolism
  • NADPH Oxidase 4 / genetics
  • NADPH Oxidase 4 / metabolism
  • NADPH Oxidases / metabolism
  • NFATC Transcription Factors / metabolism
  • Rats
  • Reactive Oxygen Species / metabolism
  • TRPC Cation Channels / genetics
  • TRPC Cation Channels / metabolism
  • TRPC6 Cation Channel / metabolism
  • Transcription Factors / metabolism

Substances

  • NADPH Oxidase 4
  • NADPH Oxidases
  • NFATC Transcription Factors
  • Nfatc1 protein, mouse
  • Reactive Oxygen Species
  • Transcription Factors
  • TRPC Cation Channels
  • TRPC6 Cation Channel
  • Trpc6 protein, mouse
  • Trpc6 protein, rat
  • NFATC1 protein, rat