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Purpose <p>Bioactive peptides, composed of short amino acid sequences, have gained attention for their ability to modulate inflammation by suppressing oxidative stress, downregulating cytokine expression, and inhibiting inflammatory mediators. This study aims to evaluate the anti-inflammatory properties of Methionine-Threonine-Histidine (MTH), a marine-derived tripeptide, using <i>in silico</i> and <i>in vitro</i> approaches.</p> Methods <p>Molecular docking simulations were employed to assess MTH's binding affinity to key inflammatory mediators (COX-2, iNOS, MMP9). Pharmacokinetic analysis predicted MTH's bioavailability and toxicity profile. <i>In vitro </i>experiments on L929 fibroblast cells stimulated with LPS investigated MTH's effects on nitric oxide production, inflammatory marker expression via western blotting, and oxidative stress levels using reactive oxygen species assays.</p> Results <p>MTH exhibited strong binding affinities to COX-2 (-6.86±0.23 kcal/mol), iNOS (-6.53±0.25 kcal/mol), and MMP9 (-7.03±0.11 kcal/mol), suggesting potent inhibitory activity. <i>In vitro</i>, MTH significantly reduced nitric oxide production (25.87 ± 0.95 μM, ***p&lt;0.001) and downregulated inflammatory markers. It also demonstrated substantial antioxidant effects (***p&lt;0.001) at 150 μM concentration.</p> Conclusions <p>The study highlights MTH as a promising anti-inflammatory agent, potentially safer than conventional NSAIDs due to its marine origin and multifunctional activity. Further <i>in vivo</i> studies are warranted to validate its therapeutic efficacy and explore applications in pharmaceutical and nutraceutical formulations for chronic inflammatory conditions.</p>

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Effects of a Bioactive Tripeptide on the Downregulation of Oxidative Stress and Inflammatory Markers in LPS-Induced Fibroblast Cells

  • 期刊论文

  • 发表日期：

    2025年6月6日

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  • Cyril Kristen,
  • Kevina Irine,
  • Sreeram Sneha,
  • Sakthivel Ragul,
  • Kandala Aashritha,
  • Akshad Balde,
  • Rasool Abdul Nazeer

摘要

Purpose

Bioactive peptides, composed of short amino acid sequences, have gained attention for their ability to modulate inflammation by suppressing oxidative stress, downregulating cytokine expression, and inhibiting inflammatory mediators. This study aims to evaluate the anti-inflammatory properties of Methionine-Threonine-Histidine (MTH), a marine-derived tripeptide, using in silico and in vitro approaches.

Methods

Molecular docking simulations were employed to assess MTH's binding affinity to key inflammatory mediators (COX-2, iNOS, MMP9). Pharmacokinetic analysis predicted MTH's bioavailability and toxicity profile. In vitro experiments on L929 fibroblast cells stimulated with LPS investigated MTH's effects on nitric oxide production, inflammatory marker expression via western blotting, and oxidative stress levels using reactive oxygen species assays.

Results

MTH exhibited strong binding affinities to COX-2 (-6.86±0.23 kcal/mol), iNOS (-6.53±0.25 kcal/mol), and MMP9 (-7.03±0.11 kcal/mol), suggesting potent inhibitory activity. In vitro, MTH significantly reduced nitric oxide production (25.87 ± 0.95 μM, ***p<0.001) and downregulated inflammatory markers. It also demonstrated substantial antioxidant effects (***p<0.001) at 150 μM concentration.

Conclusions

The study highlights MTH as a promising anti-inflammatory agent, potentially safer than conventional NSAIDs due to its marine origin and multifunctional activity. Further in vivo studies are warranted to validate its therapeutic efficacy and explore applications in pharmaceutical and nutraceutical formulations for chronic inflammatory conditions.