Investigating the effects and mechanisms of Vernonia anthelmintica (L.) willd., seed extracts on melanogenesis and vitiligo treatment based on multi-omics and network pharmacology.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Vitiligo is a depigmentation disorder characterized by the loss of functional melanocytes, leading to skin and/or hair depigmentation. Vernonia anthelmintica (L.) Willd seed extracts have been traditionally used to treat pigmentation disorders in Central Asia. However, its pharmacological mechanism for vitiligo is still unknown. AIMS OF THE STUDY: In a previous study, we isolated the Extracts (encoded as CAM-Y7) from the Vernonia anthelmintica (L.) Willd seed. This research aims to understand the molecular basis of hyperpigmentation triggered by CAM-Y7. MATERIALS AND METHODS: At first, we employed HPLC-MS to examine and measure the components in the CAM-Y7. The possible CAM-Y7-Vitiligo common targets were analyzed by network pharmacology. We explored the effects of CAM-Y7 on melanin production in melanocytes and vitiligo models induced by hydroquinone in mice and hydrogen peroxide in guinea pigs. Tissue-based metabolomics and proteomics were utilized to find varying metabolites and proteins in vitiligo mice. Through confirmatory experiments, potential therapeutic targets and molecular mechanisms were discovered. RESULTS: Twenty compounds were identified in CAM-Y7, including caffeoylquinic acids, sesquiterpenoids, and flavonoids. Network pharmacology indicated that CAM-Y7 acts on vitiligo through melanogenesis, MITF-M-regulated melanocyte development, and Tyrosine metabolism. Oral administration of CAM-Y7 progressively darkened the dorsal skin and hair of C57BL/6 mice and guinea pigs. Both Lillie staining and hematoxylin-eosin staining further demonstrated that CAM-Y7 induced melanogenesis in the epidermis and hair follicles of the animals. Multi-omics studies have shown that the Tyrosinase and MAPK pathways are important in CAM-Y7 treatment for vitiligo. Confirmatory experiments also indicated that CAM-Y7 promotes melanogenesis by upregulating MITF-induced Tyrosinase expression via the P38/MAPK-MAPKAPK2 signaling axis in melanocytes. Finally, molecular docking and SPR techniques suggested that Vernodalin is a potentially active compound in CAM-Y7. CONCLUSION: The regulation of melanogenesis by Vernonia anthelmintica (L.) Willd seed extracts might be facilitated through the activation of the P38/MAPK-MAPKAPK2 signaling axis, with Tyrosinase playing a crucial role in the melanogenesis induced by CAM-Y7.