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.