2. Academic Validation
  3. Duhuo mitigates intervertebral disc degeneration by activating PI3K/AKT to inhibit ferroptosis in nucleus pulposus cells

Duhuo mitigates intervertebral disc degeneration by activating PI3K/AKT to inhibit ferroptosis in nucleus pulposus cells

  • J Ethnopharmacol. 2026 Mar 1:358:121027. doi: 10.1016/j.jep.2025.121027.
Hao Chen 1,  Keyu Lu 2,  Xiang Li 3,  Kaiye Chen 1,  Yangcan Jin 1,  Xianpeng Chen 1,  Luyi Jiang 1,  Zhao Yang 4,  Heng Yu 5,  Zhongke Lin 6
Affiliations

Affiliations

  • 1 Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, Zhejiang Province, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, Zhejiang Province, China.
  • 2 Department of Orthopaedics, Minhang Hospital, Fudan University, Shanghai, China.
  • 3 Department of Orthopaedics, Tonglu Branch Hospital, Hangzhou First People's Hospital, No.899 Meilin Road, Tonglu, Hangzhou, 330000, Zhejiang Province, China.
  • 4 Department of Orthopaedics, The Affiliated Taizhou People's Hospital of Nanjing Medical University, Taizhou School of Clinical Medicine, Nanjing Medical University, Taizhou, China.
  • 5 Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, Zhejiang Province, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, Zhejiang Province, China. Electronic address: [email protected].
  • 6 Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, Zhejiang Province, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, Zhejiang Province, China. Electronic address: [email protected].
PMID: 41380858 DOI: 10.1016/j.jep.2025.121027
Abstract

Ethnopharmacological relevance: Duhuo (DH) is a widely used traditional Chinese medicine. It is prescribed to "dispel wind and eliminate dampness" and to relieve pain. Clinically, it is frequently administered for low back pain (LBP) associated with intervertebral disc degeneration (IDD). However, the precise molecular and cellular mechanisms by which DH may ameliorate IDD remain unclear. This limits the evidence-based translation and standardized clinical application of the medicine.

Aim of the study: To systematically assess the anti-IDD efficacy of DH by integrating network pharmacology with both in vivo and in vitro experimental validation, identify key bioactive constituents and targets, and elucidate the underlying mechanisms, with a particular focus on Ferroptosis regulation via the PI3K/Akt pathway.

Materials and methods: The chemical profile of DH was analyzed using ultra-performance liquid chromatography coupled to quadrupole orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive Orbitrap-MS). Potential targets were identified through network pharmacology and molecular docking analyses. Subsequently, pathway enrichment analysis was performed to delineate critical signaling axes. In vitro, nucleus pulposus (NP) cells were assessed to evaluate Ferroptosis, mitochondrial function, and extracellular matrix (ECM) homeostasis. The specific assays used included CCK-8 assays, fluorescence microscopy, Western blotting, flow cytometry, ROS probes, BODIPY-C11 and FerroOrange staining, and MitoTracker. In vivo, radiography (X-ray and MRI), histology, and immunohistochemistry were employed to examine structural and tissue-level alterations in rat intervertebral discs. These methods were also used to determine the efficacy of DH and its associated mechanisms.

Results: In vivo, DH intervention attenuated IDD, partially restored disc height, and improved the morphology and histoarchitecture of the NP and annulus fibrosus (AF). Mechanistically, network pharmacology and docking analyses indicated that DH may act via the PI3K/Akt signaling axis to suppress Ferroptosis in NP cells, thereby preserving mitochondrial homeostasis and mitigating ECM degradation.

Conclusion: DH may exert a protective effect against IDD by targeting the PI3K/Akt axis to reduce Ferroptosis in NP cells. These findings provide mechanistic support for using DH in IDD, based on evidence derived from traditional medicinal resources.

Keywords

Duhuo; Ferroptosis; Intervertebral disc degeneration; PI3K/AKT signaling.

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