Carnosol Attenuates Neointimal Hyperplasia in Spontaneously Hypertensive Rats: A Network Pharmacology, Molecular Docking, and Experimental Validation Study Targeting the PPAR-γ/AGTR1 Pathway.

INTRODUCTION

The study aims to investigate the effects of Carnosol on hypertensive vascular complications and its underlying mechanism.

METHODS

In vitro, A10 VSMCs were treated with angiotensin II (Ang II), Carnosol (various concentrations), and/or PPAR-γ inhibitor GW9662. Cell proliferation was detected by MTT assay, EdU staining, and immunoblotting for Proliferating Cell Nuclear Antigen (PCNA); cell migration was assessed by scratch wound assay and Transwell assay; oxidative stress was evaluated via dihydroethidium (DHE) staining, and measurements of malondialdehyde (MDA), Total Antioxidant Capacity (T-AOC), Superoxide Dismutase (SOD) activity, and NADPH oxidase 4 (NOX4) expression. Network pharmacology and molecular docking were used to predict target binding. In vivo, Spontaneously Hypertensive Rats (SHR) with carotid balloon injury were treated with Carnosol for 2 weeks, and related pathological and biochemical indices were detected.

RESULTS

Carnosol dose-dependently inhibited Ang II-induced VSMC proliferation and migration, reduced ROS and MDA levels, and restored T-AOC and SOD activity. It downregulated NOX4 and Ang II receptor type 1 (AGTR1) expression while enhancing PPAR-γ nuclear translocation and activity. These effects were reversed by GW9662. Molecular docking showed Carnosol directly bound to PPAR-γ (MM‑GBSA binding free energy: -33.18 kcal/mol). In SHR, carnosol significantly reduced the intima-to-media ratio, decreased MDA, restored T-AOC, suppressed PCNA, and upregulated PPAR-γ in injured arteries (all P < 0.05).

DISCUSSION: These findings align with PPAR-γ's role in vascular homeostasis, confirming Carnosol's direct binding to PPAR-γ to suppress Ang II/AGTR1-mediated oxidative stress and VSMC dysfunction.

CONCLUSION

Carnosol mitigates hypertension-induced VSMC dysfunction and neointimal hyperplasia by regulating the PPAR-γ/AGTR1 signaling axis.