2. Academic Validation
  3. Apigenin and its derivative modulate nitric oxide production and interfere with TLR4-M2: In-vitro and in-silico insights

Apigenin and its derivative modulate nitric oxide production and interfere with TLR4-M2: In-vitro and in-silico insights

  • Bioorg Chem. 2025 Nov:166:109069. doi: 10.1016/j.bioorg.2025.109069.
Samreen Soomro 1,  M Ahmed Mesaik 2,  Farzana Shaheen 3,  Sobia Ahsan Halim 4,  Muhammad Waqas 5,  Naira Nayeem 6,  Mashael Alanazi 7,  Ajmal Khan 8,  Ahmed Al-Harrasi 9
Affiliations

Affiliations

  • 1 Department of Pharmaceutics, Faculty of Pharmacy, Northern Border University, Rafha, Saudi Arabia.. Electronic address: tamryan.ali@nbu.edu.sa.
  • 2 Department of Medical Microbiology, Faculty of Medicine, University of Tabuk & Molecular Microbiology and Infectious Diseases Research Unit (RDIA), Deanship of Scientific Research, University of Tabuk, Tabuk 71491, Saudi Arabia.. Electronic address: mmesaik@ut.edu.sa.
  • 3 Third World Center for Science and Technology, H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
  • 4 Natural and Medical Sciences Research Center, University of Nizwa, P.O. Box 33, Birkat Al Mauz, Nizwa 616, Sultanate of Oman.. Electronic address: sobia_halim@unizwa.edu.om.
  • 5 Natural and Medical Sciences Research Center, University of Nizwa, P.O. Box 33, Birkat Al Mauz, Nizwa 616, Sultanate of Oman.
  • 6 Department of Pharmaceutical Chemistry, Faculty of Pharmacy Northern Border University, Saudi Arabia.
  • 7 Department of Pharmaceutics, Faculty of Pharmacy, Northern Border University, Rafha, Saudi Arabia.
  • 8 Natural and Medical Sciences Research Center, University of Nizwa, P.O. Box 33, Birkat Al Mauz, Nizwa 616, Sultanate of Oman.; Department of Chemical and Biological Engineering, College of Engineering, Korea University, Seongbuk-gu, 02841, Republic of Korea.
  • 9 Natural and Medical Sciences Research Center, University of Nizwa, P.O. Box 33, Birkat Al Mauz, Nizwa 616, Sultanate of Oman.. Electronic address: aharrasi@unizwa.edu.om.
PMID: 41075614 DOI: 10.1016/j.bioorg.2025.109069
Abstract

The imbalance in reactive oxygen/nitrogen species causes oxidative stress which contributes to chronic inflammation and diseases. In this study, the immunomodulatory potential of the natural product 'Apigenin (4,5,7-trihydroxyflavone)' (APG) and its derivative 'Apigenin-7-O-β-D-(6″-p-coumaroyl)-glucopyranoside' (APG-7) was elucidated through cell-based spectrophotometry, chemiluminescence, and fluorescent microscopy. Their effects were assessed on the production of superoxide anion, myeloperoxidase-dependent hypochlorite anion, intracellular oxidative stress and nitric oxide (NO). Moreover, their cellular toxicity was investigated on 3 T3 fibroblast cell line. APG significantly reduced superoxide anion (48.2 %) and hypochlorite production (IC₅₀ = 27.2 μg/mL), while APG-7 showed minimal activity in these assays. Both compounds inhibited NO production, with APG showing stronger inhibition (98 %) than APG-7 (55 %). However, APG was more cytotoxic (IC₅₀ = 4.5 μg/mL) as compared to APG-7 (∼25 μg/mL), indicating a safer profile of APG-7. NO is produced by LPS triggered activation of Toll-like Receptor 4 (TLR4), therefore in-silico molecular docking and dynamics simulation were performed to deduce the binding affinity of APG and APG-7 at TLR4/MD-2 interface. Our in-silico findings suggest that both the compounds may target TLR4/MD-2 interface to inhibit the production of NO. Overall, the results support the immunomodulatory potential of APG and APG-7, warranting further investigation.

Keywords

Inflammation; Molecular docking and molecular dynamics simulation; Oxidative stress; TLR4/MD-2.

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