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Toxics
2023 Feb 27;113:. doi: 10.3390/toxics11030226.
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Amantadine Toxicity in Apostichopus japonicus Revealed by Proteomics.
Zhao J
,
Chen J
,
Tian X
,
Jiang L
,
Cui Q
,
Sun Y
,
Wu N
,
Liu G
,
Ding Y
,
Wang J
,
Liu Y
,
Han D
,
Xu Y
.
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Amantadine exposure can alter biological processes in sea cucumbers, which are an economically important seafood in China. In this study, amantadine toxicity in Apostichopus japonicus was analyzed by oxidative stress and histopathological methods. Quantitative tandem mass tag labeling was used to examine changes in protein contents and metabolic pathways in A. japonicus intestinal tissues after exposure to 100 µg/L amantadine for 96 h. Catalase activity significantly increased from days 1 to 3 of exposure, but it decreased on day 4. Superoxide dismutase and glutathione activities were inhibited throughout the exposure period. Malondialdehyde contents increased on days 1 and 4 but decreased on days 2 and 3. Proteomics analysis revealed 111 differentially expressed proteins in the intestines of A. japonicus after amantadine exposure compared with the control group. An analysis of the involved metabolic pathways showed that the glycolytic and glycogenic pathways may have increased energy production and conversion in A. japonicus after amantadine exposure. The NF-κB, TNF, and IL-17 pathways were likely induced by amantadine exposure, thereby activating NF-κB and triggering intestinal inflammation and apoptosis. Amino acid metabolism analysis showed that the leucine and isoleucine degradation pathways and the phenylalanine metabolic pathway inhibited protein synthesis and growth in A. japonicus. This study investigated the regulatory response mechanisms in A. japonicus intestinal tissues after exposure to amantadine, providing a theoretical basis for further research on amantadine toxicity.
2022JCYJ035 Fundamental Research Projects of Science&Technology Innovation and development Plan in Yantai City, SDAIT-26- 05 Modern Agro-industry Technology Research System in Shandong Province, 2017YFC1600702 the National Key R&D Program of China
Figure 1. Histological observation of the intestine. (a) Control group. (b) Experimental group. S: plasma membrane layer; ML: muscle layer; SM: inner connective tissue layer, i.e., submucosa; M: intestinal luminal epithelium, i.e., mucosa. Black circles indicate epithelial cell necrosis. Scale bar = 100 μm.
Figure 2. Enzyme activities of (a) catalase (CAT), (b) superoxide dismutase (SOD), (c) glutathione (GSH), and (d) malondialdehyde (MDA) in the intestinal tract. Note: ** means that there is a significant difference compared with the control group (p < 0.01).
Figure 3. SDS-PAGE electrophoresis of protein samples. Note: Lanes 1–4 were control group samples; lanes 5–8 were experimental group samples.
Figure 4. Proteomics data quality control. (a) Peptide length distribution of identified peptides. (b) Distribution of the number of unique peptide segments. (c) Protein coverage distribution. (d) Protein molecular weight distribution.
Figure 5. DEPs in A. japonicus after exposure to amantadine (A) and a differential protein volcano map (B).
Figure 6. Gene Ontology enrichment analysis of differentially expressed proteins.
Figure 7. Kyoto Encyclopedia of Genes and Genomes pathways of the differentially expressed proteins.
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