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Background <p>Copper sulfate (CuSO<sub>4</sub>), a widely used industrial pollutant induces oxidative stress and inflammation, threatening aquatic organisms. This study evaluates the antioxidant and anti-inflammatory potential of the KC14 peptide, derived from <i>Cyprinus carpio</i>, in mitigating CuSO<sub>4</sub>-induced toxicity in zebrafish larvae by scavenging reactive oxygen species (ROS), regulating cytokines, and stabilizing erythrocyte membranes.</p> Methods <p>Molecular docking predicted polar interactions of KC14 peptide with inflammatory targets.Anti-inflammatory property of KC14 was evaluated using hemolytic assay. Zebrafish larvae (<i>Danio rerio</i>) were exposed to 5 µM CuSO<sub>4</sub> to induce oxidative stress, and toxicity was assessed at 24–96 hours post-fertilization (hpf). KC14 peptide was administered at concentrations ranging from 5 to 45 µM, and developmental parameters were recorded. Superoxide anion production and glutathione levels were measured to assess oxidative stress. Neutral red staining was used to evaluate macrophage response, and RT-PCR was conducted to quantify key inflammatory cytokines (TNF-α, IL-1β, IL-10) and antioxidant gene expressions.</p> Results <p>CuSO<sub>4</sub> exposure caused oxidative damage and increased mortality, while KC14 treatment (45 µM) improved survival (87%), significantly reduced ROS levels and inflammation, and enhanced antioxidant gene expression in a dose-dependent manner (<i>p</i>&#xa0;&lt;&#xa0;0.05). The peptide at 45 µM inhibited hemolysis by 76.4%, stabilized erythrocyte membranes, and downregulated pro-inflammatory cytokines while increasing IL-10 expression.</p> Conclusion <p> KC14 exhibited potent antioxidant and anti-inflammatory properties, effectively mitigating CuSO<sub>4</sub>-induced oxidative stress in zebrafish larvae. These findings indicate that KC14 effectively reduces inflammation by scavenging free radicals in CuSO<sub>4</sub>-exposed groups, highlighting its potential as a protective agent against metal-induced oxidative stress and inflammation and offering insights into its therapeutic application in environmental toxicology.</p>

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Protective Effects of KC14 Peptide from Cyprinus carpio on Copper Sulfate-Induced Toxicity in Zebrafish Larvae: Insights into Anti-inflammatory Cytokine and Glutathione Modulations

  • 期刊论文

  • 发表日期：

    2025年3月8日

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  • Madhumitha Vijayanand,
  • Praveen Kumar Issac,
  • Manikandan Velayutham,
  • Mohammed Rafi Shaik,
  • Shaik Althaf Hussain,
  • Baji Shaik,
  • Ajay Guru

摘要

Background

Copper sulfate (CuSO4), a widely used industrial pollutant induces oxidative stress and inflammation, threatening aquatic organisms. This study evaluates the antioxidant and anti-inflammatory potential of the KC14 peptide, derived from Cyprinus carpio, in mitigating CuSO4-induced toxicity in zebrafish larvae by scavenging reactive oxygen species (ROS), regulating cytokines, and stabilizing erythrocyte membranes.

Methods

Molecular docking predicted polar interactions of KC14 peptide with inflammatory targets.Anti-inflammatory property of KC14 was evaluated using hemolytic assay. Zebrafish larvae (Danio rerio) were exposed to 5 µM CuSO4 to induce oxidative stress, and toxicity was assessed at 24–96 hours post-fertilization (hpf). KC14 peptide was administered at concentrations ranging from 5 to 45 µM, and developmental parameters were recorded. Superoxide anion production and glutathione levels were measured to assess oxidative stress. Neutral red staining was used to evaluate macrophage response, and RT-PCR was conducted to quantify key inflammatory cytokines (TNF-α, IL-1β, IL-10) and antioxidant gene expressions.

Results

CuSO4 exposure caused oxidative damage and increased mortality, while KC14 treatment (45 µM) improved survival (87%), significantly reduced ROS levels and inflammation, and enhanced antioxidant gene expression in a dose-dependent manner (p < 0.05). The peptide at 45 µM inhibited hemolysis by 76.4%, stabilized erythrocyte membranes, and downregulated pro-inflammatory cytokines while increasing IL-10 expression.

Conclusion

KC14 exhibited potent antioxidant and anti-inflammatory properties, effectively mitigating CuSO4-induced oxidative stress in zebrafish larvae. These findings indicate that KC14 effectively reduces inflammation by scavenging free radicals in CuSO4-exposed groups, highlighting its potential as a protective agent against metal-induced oxidative stress and inflammation and offering insights into its therapeutic application in environmental toxicology.