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Immunoprotective effect of silybin through blocking p53-driven caspase-9-Apaf-1-Cyt c complex formation and immune dysfunction after difenoconazole exposure in carp spleen

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Abstract

As a broad-spectrum and efficient triazole fungicide, difenoconazole is widely used, which not only pollutes the environment but also exerts toxic effects on non-target organisms. The spleen plays an important role in immune protection as an important secondary lymphoid organ in carp. In this study, we assessed the protective impact of silybin as a dietary additive on spleen tissues of carp during exposure to difenoconazole. Sixty carp were separated into four groups for this investigation including control group, difenoconazole group, silybin group, and silybin and difenoconazole group. By hematoxylin-eosin staining, dihydroethidium staining, immunohistochemical staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay, quantitative real-time PCR assay, Western blot analysis, biochemical assays, and immune function indicator assays, we found that silybin could prevent difenoconazole-induced spleen tissue damage, oxidative stress, and immune dysfunction, and inhibited apoptosis of carp spleen tissue cells by suppressing the formation of p53-driven caspase-9-apoptotic protease activating factor-1-cytochrome C complex. The results suggested that silybin as a dietary additive could improve spleen tissue damage and immune dysfunction induced by difenoconazole in aquaculture carp.

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Data availability

All data generated or analyzed during this study are available from the corresponding author on reasonable request.

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Funding

This study was supported by grants from the Basic Science (Natural Science) Research Project of Higher Education of Jiangsu Province (No. 21KJB230001), Open-end Funds of Jiangsu Key Laboratory of Marine Bioresources and Environment (No. SH20221203), and the Priority Academic Program Development of Jiangsu Higher Education Institutions of China. The authors thank these funders for their financial support. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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  1. Enzhuang Pan and Yue Xin contributed equally to this study.

Authors and Affiliations

  1. Jiangsu Key Laboratory of Marine Bioresources and Environment, Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Marine Pharmaceutical Resources Development Engineering Research Center, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, College of Pharmacy, Jiangsu Ocean University, Lianyungang, 222005, China

    Enzhuang Pan, Yue Xin, Xueqing Li, Kaixin Ping, Xing Li, Ying Sun, Xuhui Xu & Jingquan Dong

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  1. Enzhuang Pan
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  2. Yue Xin
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Contributions

Enzhuang Pan: data curation, formal analysis, methodology, and writing—original draft. Yue Xin: data curation, formal analysis, investigation, and validation. Xueqing Li: data curation, formal analysis, investigation, and validation. Kaixin Ping: investigation, resources, software, and supervision. Xing Li: formal analysis, methodology, and supervision. Ying Sun: data curation and validation. Xuhui Xu: data curation and validation. Jingquan Dong: conceptualization, project administration, funding acquisition, validation, and writing—review and editing. All authors read and approved the final manuscript.

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Correspondence to Jingquan Dong.

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All animal experiments were approved by the Institutional Animal Ethics Committee of Jiangsu Ocean University, and animal care was in accordance with the institutional guidelines.

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Pan, E., Xin, Y., Li, X. et al. Immunoprotective effect of silybin through blocking p53-driven caspase-9-Apaf-1-Cyt c complex formation and immune dysfunction after difenoconazole exposure in carp spleen. Environ Sci Pollut Res 31, 19396–19408 (2024). https://doi.org/10.1007/s11356-024-32392-0

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