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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

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Abstract

Background

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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.

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Data Availability

Data are included within the manuscript or supplementary file. Further details are available from the corresponding author on reasonable request.

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Acknowledgements

The authors gratefully acknowledge Dr. Ghulam Jelani, Department of Botany, University of Peshawar, for taxonomic identification and verification of Plant Lilium polyphyllum.

Funding

No funding was received for this study.

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Authors and Affiliations

  1. Department of Pharmacy, Shaheed Benazir Bhutto University, Sheringal, Khyber Pakhtunkhwa, Dir 18050, Pakistan

    Haider Ali, Shafiq Ur Rahman & Umair Sayad

  2. Department of Biotechnology, Yeungnam University, Gyeongsan, 38541, Republic of Korea

    Atif Ali Khan Khalil

  3. Department of Pharmacognosy, Faculty of Pharmaceutical and Allied Health Sciences, Lahore College for Women University, Lahore, 54000, Pakistan

    Atif Ali Khan Khalil

  4. Faculty of Pharmacy, Capital University of Science & Technology, Islamabad, 44000, Pakistan

    Muzaffar Abbas

  5. Department of Pharmacy, CECOS University of Information Technology and Emerging Sciences, Peshawar, Khyber Pakhtunkhwa, 25000, Pakistan

    Muhammad Shahid

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Contributions

HA: Conducted the experiments, interpreted the results, and drafted the manuscript. SUR: Supervised the overall experiments, provided the necessary facilities and resources. US: Assisted with data tabulation and statistics. AAKK: Assisted in phytochemical analysis, molecular docking, and proofreading. MA: Assisted with the in vivo experiments. MS: Contributed to manuscript editing and proofreading. All authors reviewed and approved the final manuscript.

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Correspondence to Shafiq Ur Rahman or Atif Ali Khan Khalil.

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Ali, H., Ur Rahman, S., Sayad, U. et al. Phytochemical Profiling and Evaluation of the Anti-inflammatory and Antinociceptive Effects of Lilium polyphyllum Bulb Using In Silico, In Vitro and In Vivo Models. J Pharm Innov 21, 53 (2026). https://doi.org/10.1007/s12247-025-10267-2

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