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Background <p><i>Lilium Polyphyllum </i>D. Don ex Royle (Himalayan lily) has long been valued in traditional medicine for managing pain, rheumatism, bronchitis, and intermittent fever. Despite its ethnomedicinal relevance, robust scientific evidence supporting its anti-inflammatory and analgesic effects remains limited.</p> Objectives <p>This study aims to validate the traditional anti-inflammatory and analgesic uses of <i>L. polyphyllum</i> bulb by characterizing its phytochemical constituents and elucidating the underlying mechanisms of action through integrated in silico, in vitro and in vivo approaches.</p> Methods <p>LC-MS/MS and GC-MS analyses were conducted for the most bioactive chloroform (Chl-fr) and ethyl acetate fractions (EtOAc-fr) respectively. Toxicity was assessed via fibroblast viability, brine shrimp lethality, and acute in vivo testing up to 2400&#xa0;mg/kg, supported by SwissADMET predictions. Anti-inflammatory effects were determined using chemiluminescence and carrageenan-induced paw edema models, while antinociceptive activity was evaluated through acetic acid-induced writhing and hot plate assays. Molecular docking was performed to explore interactions of identified compounds with myeloperoxidase (MPO) and P2X purinergic receptor 7 (P2X7R).</p> Results <p>LCMS/MS and GC-MS analyses of Chl-fr and EtOAc-fr identified 55 bioactive compounds, including phenolic acids, glycosides, alkaloids, flavonoids, and fatty acid esters. The Cr. MeOH-Ext and fractions were non-toxic in fibroblast cells, brine shrimp assay, and in vivo up to 2400&#xa0;mg/kg, with SwissADMET predictions further supporting their safety profile. The in vitro anti-inflammatory results revealed that Chl-fr, EtOAc-fr, and <i>n</i>-Hex-fr exhibited significant inhibition of oxidative burst for whole blood. In carrageenan-induced anti-inflammatory test, significant decrease in paw edema was observed at 50 and 100&#xa0;mg/kg for Cr. MeOH-Ext and at 100&#xa0;mg/kg for <i>n</i>-Hex-fr, Chl-fr and EtOAc-fr. In abdominal constriction test, EtOAc-fr at 50 and 100&#xa0;mg/kg significantly reduced abdominal writhing followed by <i>n</i>-Hex-fr, Cr. MeOH-Ext, and Chl-fr. In hot plate test, the latency time was significantly increased at 100&#xa0;mg by EtOAc-fr followed by Chl-fr, and <i>n</i>-Hex-fr. Molecular docking analysis revealed that the identified compounds exhibit strong binding affinities with MPO and P2X7R, further supporting their potential anti-inflammatory and antinociceptive mechanisms.</p> Conclusion <p>To our knowledge, this study provides the first comprehensive characterization of the phytochemical profiles of chloroform and ethyl acetate fractions of <i>L. polyphyllum</i> bulb, demonstrating significant analgesic and anti-inflammatory potential that scientifically validate the ethnomedicinal applications in pain and inflammation.</p>

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Phytochemical Profiling and Evaluation of the Anti-inflammatory and Antinociceptive Effects of Lilium polyphyllum Bulb Using In Silico, In Vitro and In Vivo Models

  • 期刊论文

  • 发表日期：

    2025年12月3日

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  • Haider Ali,
  • Shafiq Ur Rahman,
  • Umair Sayad,
  • Atif Ali Khan Khalil,
  • Muzaffar Abbas,
  • Muhammad Shahid

摘要

Background

Lilium Polyphyllum D. Don ex Royle (Himalayan lily) has long been valued in traditional medicine for managing pain, rheumatism, bronchitis, and intermittent fever. Despite its ethnomedicinal relevance, robust scientific evidence supporting its anti-inflammatory and analgesic effects remains limited.

Objectives

This study aims to validate the traditional anti-inflammatory and analgesic uses of L. polyphyllum bulb by characterizing its phytochemical constituents and elucidating the underlying mechanisms of action through integrated in silico, in vitro and in vivo approaches.

Methods

LC-MS/MS and GC-MS analyses were conducted for the most bioactive chloroform (Chl-fr) and ethyl acetate fractions (EtOAc-fr) respectively. Toxicity was assessed via fibroblast viability, brine shrimp lethality, and acute in vivo testing up to 2400 mg/kg, supported by SwissADMET predictions. Anti-inflammatory effects were determined using chemiluminescence and carrageenan-induced paw edema models, while antinociceptive activity was evaluated through acetic acid-induced writhing and hot plate assays. Molecular docking was performed to explore interactions of identified compounds with myeloperoxidase (MPO) and P2X purinergic receptor 7 (P2X7R).

Results

LCMS/MS and GC-MS analyses of Chl-fr and EtOAc-fr identified 55 bioactive compounds, including phenolic acids, glycosides, alkaloids, flavonoids, and fatty acid esters. The Cr. MeOH-Ext and fractions were non-toxic in fibroblast cells, brine shrimp assay, and in vivo up to 2400 mg/kg, with SwissADMET predictions further supporting their safety profile. The in vitro anti-inflammatory results revealed that Chl-fr, EtOAc-fr, and n-Hex-fr exhibited significant inhibition of oxidative burst for whole blood. In carrageenan-induced anti-inflammatory test, significant decrease in paw edema was observed at 50 and 100 mg/kg for Cr. MeOH-Ext and at 100 mg/kg for n-Hex-fr, Chl-fr and EtOAc-fr. In abdominal constriction test, EtOAc-fr at 50 and 100 mg/kg significantly reduced abdominal writhing followed by n-Hex-fr, Cr. MeOH-Ext, and Chl-fr. In hot plate test, the latency time was significantly increased at 100 mg by EtOAc-fr followed by Chl-fr, and n-Hex-fr. Molecular docking analysis revealed that the identified compounds exhibit strong binding affinities with MPO and P2X7R, further supporting their potential anti-inflammatory and antinociceptive mechanisms.

Conclusion

To our knowledge, this study provides the first comprehensive characterization of the phytochemical profiles of chloroform and ethyl acetate fractions of L. polyphyllum bulb, demonstrating significant analgesic and anti-inflammatory potential that scientifically validate the ethnomedicinal applications in pain and inflammation.