Bioactive-rich Mangifera indica leaf extracts alleviate inflammation, pain, and fever via suppression of pro-inflammatory cytokines and activation of Nrf2 pathways.

Leaves of Mangifera indica (mango) are traditionally used to alleviate inflammation and pain, yet their bioactive chemical profiles and mechanisms of action remain underexplored. Given the growing demand for safer, plant-derived therapeutics as alternatives to conventional anti-inflammatory and analgesic drugs with known adverse effects, a deeper understanding of the bioactive composition of M. indica leaf is both timely and necessary. This study investigates the metabolomic composition and therapeutic potential of leaf extracts from two cultivars-Zebda and Alphonso. Aqueous, ethanol, n-butanol, and ethyl acetate extracts were analyzed using UHPLC-ESI-LIT-Orbitrap-MS and multivariate tools. In vivo assays were conducted to evaluate anti-inflammatory, analgesic, and antipyretic activities using biochemical, histopathological, and immunohistochemical assessments. Metabolomic profiling identified 48 compounds, including phenolics, flavonoids, xanthones, and terpenes. EtOAc extracts were enriched in methyl gallate, maclurin derivatives, and iriflophenone compounds, while aqueous and EtOH extracts contained higher levels of mangiferin, isomangiferin, and quinic acid. Biological evaluations revealed that the EtOAc and H2O extracts from both cultivars significantly reduced pro-inflammatory markers such as TNF-α, IL-1β, COX-2, and PGE2. Histological analysis confirmed decreased dermal inflammation and edema. Notably, these extracts also enhanced Nrf2 expression, a key regulator of oxidative stress response. Analgesic and antipyretic tests showed dose- and time-dependent efficacy, with EtOAc extracts demonstrating the longest-lasting pain relief and the fastest fever reduction. PLS regression analysis linked the metabolite profiles of mango extracts to their bioactivities, identifying key metabolites associated with anti-inflammatory effects viz. iriflophenone-O-hexoside, apigenin-C-hexoside, analgesic activity viz. dihydroxy-octadecenoic acid, quercetin, and antipyretic action viz. maclurin-O-galloylhexoside, mangiferin/isomangiferin, galloyl-O-hexose, quercetin-O-hexoside, unidentified triterpene, and O-hydroxybenzoyl mangiferin. By integrating metabolomic profiling with functional in vivo assays, this work contributes to the rational validation of traditional medicinal uses of mango leaves and supports their potential as a sustainable source of bioactives for inflammation, pain, and fever management.