Abstract | ETHNOPHARMACOLOGICAL RELEVANCE: Centella asiatica (L.) Urban is an ethnobotanical herb. The main bioactive components of Centella asiatica are pentacyclic triterpenoid glycosides, namely asiaticoside and hydroxyasiaticoside. Asiaticoside possess a diverse array of pharmacological properties, such as wound-healing, anti-inflammatory, antioxidant, anti-allergic, antidepressant, anxiolytic, anti-fibrotic, antibacterial, anti-arthritic, anti- tumor, and immunomodulatory activities. AIM OF THE STUDY: The purpose of this investigation is to explore potential therapeutic interventions for the delayed healing of wounds in diabetic patients (DW) facilitated by Asiaticoside- Nitric Oxide. To clarify the key molecular mechanism of miRNA-21-5p in DW wound repair and to deepen the understanding of DW disease pathogenesis. MATERIALS AND METHODS: Firstly, miRNA microarray technology, bioinformatics, and RT-qPCR were used to analyze DW patients' and normal controls' skin tissue samples. Secondly, in order to investigate the role of miRNA-21-5p, a hyperglycemic model was established using HaCaT cells. Overexpressing as well as interfering HaCaT cell lines were constructed by lentiviral infection to further explore the proliferative and migratory effects of Asiaticoside- Nitric Oxide. The next step was to search for potential target genes of miRNA-21-5p and verify them with dual- luciferase reporter assay. Finally, the expression levels of target genes and proteins were detected through the utilization of RT-qPCR and Western blotting under the influence of Asiaticoside- Nitric Oxide. RESULTS: A library of miRNAs and target genes expressed explicitly in DW patients and rats was established. The study confirmed the upregulation of miRNA-21-5p in DW patients and identified its involvement in signaling pathways related to chronic ulcer wound repair. Overexpression of LV-miRNA-21-5p significantly promoted cell proliferation, while treatments of Asiaticoside-Low dose (AC-L) and Asiaticoside-Medium dose (AC-M) enhanced proliferation and migration, particularly when combined with nitroprusside (SNP). Further analysis revealed potential target genes of miRNA-21-5p, such as TGF-β1, SMAD7, and TIMP3. Their interaction with miRNA-21-5p was confirmed through dual luciferase assays. The study found that anti-DW drugs increased the expression of TGF-β1 and SMAD7 while inhibiting TIMP3 expression in a high- glucose environment. CONCLUSIONS: The research concluded that miRNA-21-5p plays a crucial role in the delayed healing of diabetic wounds, and that the combination treatment of AC + SNP shows promise in promoting wound healing in DW rats. Target genes, including TGF-β1, SMAD7, and TIMP3, may contribute to the regulatory mechanisms involved in diabetic wound healing. These findings provide valuable insights for developing novel therapeutic approaches for DW.
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Authors | Ye Liu, Jiufeng Zhao, Xingrui Mu, Junyu Deng, Xingqian Wu, Wenjie He, Yiqiu Liu, Rifang Gu, Felicity Han, Xuqiang Nie |
Journal | Journal of ethnopharmacology
(J Ethnopharmacol)
Vol. 319
Issue Pt 2
Pg. 117266
(Jan 30 2024)
ISSN: 1872-7573 [Electronic] Ireland |
PMID | 37783408
(Publication Type: Journal Article)
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Copyright | Copyright © 2023 Elsevier B.V. All rights reserved. |
Chemical References |
- Transforming Growth Factor beta1
- asiaticoside
- Nitric Oxide
- MicroRNAs
- Luciferases
- TIMP3 protein, human
- Tissue Inhibitor of Metalloproteinase-3
- SMAD7 protein, human
- Smad7 Protein
- MIRN21 microRNA, human
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Topics |
- Humans
- Rats
- Animals
- Transforming Growth Factor beta1
(metabolism)
- Nitric Oxide
(metabolism)
- Wound Healing
- Signal Transduction
- MicroRNAs
(genetics)
- Diabetes Mellitus
- Luciferases
- Tissue Inhibitor of Metalloproteinase-3
(metabolism, pharmacology)
- Smad7 Protein
(metabolism, pharmacology)
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