Yiqi Xugu HeJi restores cartilage metabolic homeostasis via AKT1-Thr473 activation in osteoarthritis

Phytomedicine. 2025 Nov 25:148:157228. doi: 10.1016/j.phymed.2025.157228. Epub 2025 Sep 5.

Abstract

Background: Osteoarthritis (OA) is a degenerative joint disease marked by cartilage degradation, inflammation, and changes in subchondral bone structure. Despite its widespread use in treating OA, the mechanisms of traditional Chinese medicine (TCM) are not well understood.

Purpose: This study aimed to explore the bioactive components and therapeutic mechanism of Yiqi Xugu HeJi (YQXGHJ), a classic TCM prescription, in alleviating OA progression.

Methods: A destabilization of the medial meniscus (DMM)-induced OA rat model was established to evaluate the protective effects of YQXGHJ. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) identified serum-absorbed compounds, while network pharmacology and molecular docking predicted potential targets. Cellular and molecular validation, including CCK-8, EdU, flow cytometry, qPCR, Western blotting, immunofluorescence, and Transwell assays, were performed in IL-1β-induced chondrocyte inflammation models. Key signaling pathways were verified using the AKT inhibitor MK2206.

Results: YQXGHJ significantly alleviated cartilage degeneration and subchondral bone sclerosis in DMM rats. In vitro, YQXGHJ-containing serum promoted chondrocyte proliferation, suppressed apoptosis, and restored the balance between anabolic (COL2A1, SOX9) and catabolic (MMP13, IL-6) markers. Network pharmacology and docking pinpointed AKT1 as a central target, with KEGG analysis emphasizing the PI3K-AKT pathway. Western blot and immunofluorescence confirmed that YQXGHJ activated PI3K/AKT signaling and enhanced phosphorylation at the AKT1-T473 site. Inhibition of AKT1 phosphorylation by MK2206 abolished these protective effects, confirming pathway specificity.

Conclusion: YQXGHJ mitigates OA progression by reestablishing cartilage metabolic balance through the activation of the PI3K/AKT signaling pathway, specifically enhancing AKT1-T473 phosphorylation. This study provides a pharmacological and mechanistic foundation for the clinical use of YQXGHJ in treating OA.

Keywords: AKT1 phosphorylation; Chondrocyte metabolic homeostasis; Osteoarthritis; Traditional Chinese medicine; Yiqi Xugu HeJi.

MeSH terms

  • Animals
  • Cartilage* / drug effects
  • Cartilage* / metabolism
  • Cartilage, Articular* / drug effects
  • Cartilage, Articular* / metabolism
  • Cell Proliferation / drug effects
  • Chondrocytes / drug effects
  • Chondrocytes / metabolism
  • Disease Models, Animal
  • Drugs, Chinese Herbal* / pharmacology
  • Heterocyclic Compounds, 3-Ring / pharmacology
  • Homeostasis / drug effects
  • Male
  • Molecular Docking Simulation
  • Osteoarthritis* / drug therapy
  • Osteoarthritis* / metabolism
  • Phosphorylation / drug effects
  • Proto-Oncogene Proteins c-akt* / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction / drug effects

Substances

  • Proto-Oncogene Proteins c-akt
  • Drugs, Chinese Herbal
  • Akt1 protein, rat
  • Heterocyclic Compounds, 3-Ring
  • MK 2206