Abstract
Approximately 50 million people are suffering from epilepsy worldwide. Corals have been used for treating epilepsy in traditional Chinese medicine, but the mechanism of this treatment is unknown. In this study, we analyzed the transcriptome of the branching coral Acropora digitifera and obtained its Kyoto Encyclopedia of Genes and Genomes (KEGG), EuKaryotic Orthologous Groups (KOG) and Gene Ontology (GO) annotation. Combined with multiple sequence alignment and phylogenetic analysis, we discovered three polypeptides, we named them AdKuz1, AdKuz2 and AdKuz3, from A. digitifera that showed a close relationship to Kunitz-type peptides. Molecular docking and molecular dynamics simulation indicated that AdKuz1 to 3 could interact with GABAA receptor but AdKuz2–GABAA remained more stable than others. The biological experiments showed that AdKuz1 and AdKuz2 exhibited an anti-inflammatory effect by decreasing the aberrant level of nitric oxide (NO), IL-6, TNF-α and IL-1β induced by LPS in BV-2 cells. In addition, the pentylenetetrazol (PTZ)-induced epileptic effect on zebrafish was remarkably suppressed by AdKuz1 and AdKuz2. AdKuz2 particularly showed superior anti-epileptic effects compared to the other two peptides. Furthermore, AdKuz2 significantly decreased the expression of c-fos and npas4a, which were up-regulated by PTZ treatment. In addition, AdKuz2 reduced the synthesis of glutamate and enhanced the biosynthesis of gamma-aminobutyric acid (GABA). In conclusion, the results indicated that AdKuz2 may affect the synthesis of glutamate and GABA and enhance the activity of the GABAA receptor to inhibit the symptoms of epilepsy. We believe, AdKuz2 could be a promising anti-epileptic agent and its mechanism of action should be further investigated.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
This work was supported by University of Macau was funded by the Science and Technology Development Fund, Macau SAR (File no. FDCT0058/2019/A1 and 0016/2019/AKP), University of Macau (Ref No: MYRG2020-00183-ICMS, MYRG2022-00263-ICMS and CPG2022-00023-ICMS) and Shenzhen-Hong Kong-Macao Science and Technology Innovation Project (Category C) (Ref No: EF038/ICMS-LMY/2021/SZSTIC), Shenzhen Science and Technology Innovation Committee. The Hong Kong Polytechnic University (Project ID. P0006304). The Environmental and Conservation Fund of Hong Kong (Grant no. 34/2019), and Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong laboratory (Guangzhou) (Grant nos. GMl2019ZD0404, SMSEGl20SC02). This work was performed in part at the high performance computing cluster (HPCC) which is supported by information and communication technology office (ICTO) of the University of Macau.
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HBC engaged in conceptualization and bioinformatics analysis; HBC, SWI and CTT conducted the bioassay and analyzed the data. HBC and AKK prepared the manuscript. JWQ and SMY reviewed and edited the manuscript. All authors have read and agreed to the published version of the manuscript.
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Chen, H., Siu, S.W.I., Wong, C.T.T. et al. Anti-epileptic Kunitz-like peptides discovered in the branching coral Acropora digitifera through transcriptomic analysis. Arch Toxicol 96, 2589–2608 (2022). https://doi.org/10.1007/s00204-022-03311-4
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DOI: https://doi.org/10.1007/s00204-022-03311-4