Rhoifolin inhibits ferroptosis and alleviates pulmonary arterial hypertension via the TNF-α/TNF-R1/CASP8/CASP3 pathway

  • Phytomedicine. 2026 Jun:155:158095. doi: 10.1016/j.phymed.2026.158095.
Shaodong Deng  1 ,  Yaoxian Lao  2 ,  Jincheng Xie  2 ,  Zhizhong Pang  2 ,  Qingxian Xie  2 ,  Mengyuan Xiao  3 ,  Jianying Chen  4
Affiliations
  • 1. Guangdong Medical University, Dongguan, Guangdong 523808, China; Scientific Research Platform, The Second School of Clinical Medicine, Guangdong Medical University, Dongguan, Guangdong 523808, China; The First Dongguan Affiliated Hospital, Guangdong Medical University, Dongguan, Guangdong 523710, China. Electronic address: [email protected].
  • 2. Guangdong Medical University, Dongguan, Guangdong 523808, China; Scientific Research Platform, The Second School of Clinical Medicine, Guangdong Medical University, Dongguan, Guangdong 523808, China; Department of Structural Heart Disease, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, China.
  • 3. Guangdong Medical University, Dongguan, Guangdong 523808, China; Department of Structural Heart Disease, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, China.
  • 4. Guangdong Medical University, Dongguan, Guangdong 523808, China; Department of Structural Heart Disease, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, China. Electronic address: [email protected].
PMID: 41895094 DOI: 10.1016/j.phymed.2026.158095
Abstract

Background: Pulmonary arterial hypertension (PAH) is a fatal disease driven by progressive vascular remodeling. Emerging evidence links Ferroptosis, an iron-dependent form of regulated cell death, to PAH pathogenesis, suggesting that inhibiting Ferroptosis is a promising therapeutic strategy. Rhoifolin (ROF), a natural flavonoid with multiple bioactivities, has not been studied in PAH.

Purpose: This study aimed to systematically investigate the therapeutic potential of ROF against PAH, focusing on its inhibition of Ferroptosis and the underlying molecular mechanisms.

Methods: An integrative approach was employed. Network pharmacology predicted common targets of ROF, Ferroptosis, and PAH. Molecular docking and dynamics simulations assessed binding stability. Predictions were rigorously validated in vitro using Erastin-stimulated rat pulmonary arterial smooth muscle cells (rPASMCs) and in vivo using a monocrotaline-induced PAH rat model.

Results: Bioinformatics analysis identified 60 common targets and highlighted the TNF-α/TNF-R1/CASP8/CASP3 axis within the lipid and atherosclerosis pathway as a key mechanism. In vitro, ROF directly rescued rPASMCs from Erastin-induced Ferroptosis, confirming its cell-protective effect. In vivo, ROF treatment ameliorated hemodynamic and remodeling indices, reduced pulmonary Ferroptosis markers (Fe2+, MDA), and restored anti-ferroptotic defenses (GSH, GPX4). Concurrently, it downregulated the protein levels of the TNF-α/TNF-R1/CASP8/CASP3 axis. Computational studies confirmed stable binding of ROF to key targets.

Conclusions: This study is the first to demonstrate that ROF alleviates PAH by inhibiting Ferroptosis, a mechanism linked to modulation of the TNF-α/TNF-R1/CASP8/CASP3 signaling axis. Our findings position ROF as a novel multi-target candidate for PAH therapy.

Keywords
Ferroptosis; Molecular docking; Molecular dynamics simulation; Network pharmacology; Pulmonary arterial hypertension; Rhoifolin.
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