Baicalein Induces Hepatic Stellate Cell Senescence and Attenuates Liver Fibrosis via CEBPZ/p53/HK2-Mediated Glycolysis Inhibition

  • Phytomedicine. 2025 Sep 3:148:157222. doi: 10.1016/j.phymed.2025.157222.
Shuling Chen  1 ,  Ruiqi Li  2 ,  Caiyun Zhang  3 ,  Jun Guo  4 ,  Anyin Yang  5 ,  Dawei Yang  6 ,  Tianhao Mao  3 ,  Hongli Liu  2 ,  Hao Ren  4 ,  Zihao Liang  7 ,  Wei Chen  7 ,  Changhua Yi  7 ,  Qingfang Xiong  4 ,  Dandan Yin  8 ,  Xixuan Wang  9 ,  Yongfeng Yang  10
Affiliations
  • 1. Department of Hepatology, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China; Department of Pharmacy, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing, 210000, China.
  • 2. Department of Hepatology, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China; Medical School, Southeast University, Nanjing 210000, Jiangsu, China.
  • 3. Nanjing Key Laboratory of Hepatology, Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China.
  • 4. Department of Hepatology, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China.
  • 5. Department of Pharmacy, Gaochun Hospital Affiliated to Jiangsu University, Nanjing 210000, Jiangsu, China.
  • 6. Department of Pharmacy, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing, 210000, China.
  • 7. Clinical Research Centre, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China.
  • 8. Clinical Research Centre, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China. Electronic address: [email protected].
  • 9. Department of Hepatology, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China; Medical School, Southeast University, Nanjing 210000, Jiangsu, China. Electronic address: [email protected].
  • 10. Department of Hepatology, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China; Nanjing Key Laboratory of Hepatology, Nanjing University of Chinese Medicine, Nanjing 210000, Jiangsu, China. Electronic address: [email protected].
PMID: 41075517 DOI: 10.1016/j.phymed.2025.157222
Abstract

Background: Liver fibrosis progresses to life-threatening cirrhosis, yet effective therapies remain limited. Baicalein (BA), a bioactive flavonoid, shows anti-tumor potential but its anti-liver-fibrotic efficacy is completely unexplored.

Purpose: This study aimed to investigate the anti-liver-fibrotic efficacy of BA and elucidate its underlying molecular mechanisms.

Methods: We employed carbon tetrachloride (CCl4)-induced fibrotic mice and transforming growth factor-β1-activated LX-2 human hepatic stellate cells (HSC). Comprehensive assessments included: serum biochemical analysis, histopathological examination, immunofluorescence/immunohistochemistry, the analysis of cell senescence staining, glycolytic flux assays, the assessment of the target of BA and the validation of liquid-liquid phase separation (LLPS).

Results: BA significantly attenuated liver injury (AST and AST decreasing, P < 0.001), inflammation (interleukin-6, interleukin-8 and tumor necrosis factor-alpha decreasing, P < 0.001) and Collagen deposition (P < 0.001) in fibrotic mice. In activated HSCs, BA suppressed cellular activation while inducing senescence through G1-phase cell cycle arrest (P < 0.001) and glycolysis inhibition (lactate decreasing: P < 0.001). Mechanistically, BA directly bound to CEBPZ, suppressing its LLPS and subsequently enhancing TP53 transcription. This activation of the p53/HK2 axis drove HSCs senescence and attenuated fibrosis. Critically, pharmacological inhibition of p53 with pifithrin-α hydrobromide abolished BA's anti-fibrotic effects. BA's anti-fibrotic efficacy was entirely lost in CEBPZ-knockdown models (P > 0.05), confirming the essentiality and novelty of this target.

Conclusion: Our study revealed, for the first time, that BA ameliorated liver fibrosis by directly targeting CEBPZ, disrupting its LLPS, and activating the p53/HK2 axis to induce HSC senescence. This work uncovered a novel plant-derived therapeutic strategy targeting a previously unrecognized CEBPZ-LLPS/p53/HK2 mechanism in hepatic fibrosis.

Keywords
Baicalein; CEBPZ; Hepatic stellate cells; Liver fibrosis; Senescence; p53.
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