Study on the action mechanism of the peptide compounds of Wuguchong on diabetic ulcers, based on UHPLC-Q-TOF-MS, network pharmacology and experimental validation.

Journal of ethnopharmacology(2022)

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摘要
ETHNOPHARMACOLOGICAL RELEVANCE:Diabetic foot ulcers mainly refer to people who are initially diabetic and do not have peripheral neuropathy or peripheral vascular disease, but have developed foot infection, septicemia, and ulceration. Diabetic trauma disease is characterized by high sugar and very slow wound healing, which is the reason why some patients with severe diabetic trauma require amputation. Prolonged hyperglycemia can lead to changes in bodily functions and endocrine changes, which can lead to permeability damage of epidermal tissue structure, microvascular damage and, in more severe cases, nerve damage, which are also the main causes of diabetic trauma. Small molecule peptides have various biological activities, such as: lowering blood pressure, antibacterial and wound healing activities, etc. It is a drug recorded in classical Chinese medicine, it is safer to use natural active peptides to treat wounds compared to the listed drugs, and there are no side effects in its use.The wound healing effect of Wuguchong dry product has been confirmed but the mechanism is still unclear, whether it is related to the small molecule active peptides contained in it remains to be studied. AIM OF STUDY:Objective To investigate the potential mechanism of the peptide compounds of Wuguchong (PCW) on diabetic wound healing and the relevant targets in the pathway associated with the treatment of diabetic ulcers using a systematic pharmacological and pharmacological experimental validation approach. METHODS:1) PCW was prepared by enzymatic digestion of TCMW and analyzed by UHPLC-Q-TOF-MS. 2) Further screening of the active chemical components of PCW using PubChem, Swiss Target Prediction data. 3) Prediction of its targets using Drug Bank, CTD, and Genecards databases. 4) Construct protein/gene interactions network diagrams for PCWs acting by using Cytoscape 3.7.0 software. 5) GO and KEGG analysis of PCW targets were performed by David database. 6) Validated by AO/EB staining, scratching and in vitro tube formation methods. 7) Explored the mechanism of PCW to promote diabetic wound healing by protein blotting and immunohistochemical detection of relevant protein expression. RESULTS:and finally: 1) After the above screening, 81 active ingredients of PCW and 94 targets acting on diabetic ulcers were obtained. 2) 30 biological processes, 30 cellular compositions and 30 molecular functions were obtained by GO analysis; 28 signaling pathways were obtained by KEGG analysis. 3) The results of AO/EB staining assay, scratch assay and in vitro tube-forming assay showed that PCW has significant pro-vascular endothelial cell proliferation and pro-angiogenic effects in vitro. CONCLUSIONS:The results of this study confirmed the effect of the PCW in treating diabetic ulcers to a certain extent, and further revealed its mechanism of action in depth, which provides a new reference for the next step of Chinese medicine in treating diabetic ulcers.
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