Abstract
The aqueous ethanol extract of Periplaneta americana (PAE) dramatically stimulates wound healing by promoting the migration of keratinocytes. The aims of this study were to identify the active oligopeptides present in PAE that promote wound healing and to elucidate their underlying molecular mechanisms. For the detection of cell migration and proliferation in vitro, Transwell assays, wound healing assays, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed on the human keratinocyte cell line HaCaT. Western blot analysis was performed on the HaCaT cells to detect the protein levels of SMAD family member 3 (Smad3) and phosphorylated Smad3 (P-Smad3). Immunofluorescence analysis was performed on the HaCaT cells to detect the degree of Smad3 translocation into the nucleus. Three oligopeptides were purified from PAE via a series of chromatographic techniques, and their individual effects on HaCaT cell migration were evaluated using wound healing assays. The purification techniques employed were gel filtration chromatography, C8 reversed-phase column chromatography, and reversed-phase high-performance liquid chromatography (RP-HPLC). The amino acid sequences of the oligopeptides (LAKF, PYAHF, and PQLSY) were identified via HPLC–tandem mass spectrometry (HPLC–MS/MS) and Edman degradation. The oligopeptide with the PYAHF sequence significantly promoted HaCaT cell migration. In addition, this oligopeptide promoted the nuclear translocation of P-Smad3 by activating the Smad3 signaling pathway. The findings of this study provide a foundation for the future development of peptide drugs for promoting wound healing.
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This work was supported by the National Natural Science Foundation of China (no. 81973575), and 2022 National College Students’ innovation training program of Chengdu university (no. 202211079030).
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Writing—Original Draft, Data Curation: YL. Writing—Original Draft: XD. Data Curation: HH. Data Curation: JZ. Data Curation: YP. Conceptualization, Investigation, Methodology: HY. Conceptualization, Investigation, Methodology, Supervision: QS.
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Liu, Y., Dai, X., Hu, H. et al. Isolation and Purification of an Oligopeptide from Periplaneta americana and Its Mechanism of Promoted Wound Healing. Int J Pept Res Ther 29, 65 (2023). https://doi.org/10.1007/s10989-023-10536-z
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DOI: https://doi.org/10.1007/s10989-023-10536-z