Unveiling the wound-healing potential of Pisum sativum aerial biomass through integrated bioassay-guided, LC-MS/MS, and network pharmacology approaches.

BACKGROUND: While Pisum sativum L. is primarily valued for its seeds, its aerial parts have received limited attention. This study aimed to explore the anti-inflammatory and wound-healing potential of pea leaves and stems to promote their broader utilization. METHODS: A bioassay-guided strategy was applied to various fractions of P. sativum aerial parts to evaluate their immunomodulatory activity using leukocyte accumulation assay. Active fractions were further assessed in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages for their effects on TNF-α gene expression, interleukin-6 (IL-6), and nitric oxide production. The bioactive petrol eum ether (PSP) and ethyl acetate (PSE) fractions were further analyzed through GC-MS analysis, LC-MS/MS analysis, and chromatographic purification. The purified compounds were evaluated in human monocytes for their effects on IL-6 secretion and intracellular reactive oxygen species (ROS) production, and in HaCaT keratinocytes for their wound-healing potential using a scratch assay. Network pharmacology and molecular docking analyses were employed as in-silico tools to predict key targets and mechanistic pathways. RESULTS: Both PSE and PSP fractions promoted leukocyte accumulation and significantly decreased TNF-α expression, IL-6, and nitric oxide production levels. LC-MS/MS analysis of PSE revealed 42 compounds, predominantly flavonoids and phenolics, potentially accounting for its anti-inflammatory activity. Among seven isolated constituents, pisatin exhibited superior activity by markedly suppressing IL-6 and ROS in human monocytes and promoting 73% wound closure in keratinocytes within 48 h. CONCLUSIONS: These findings establish P. sativum aerial parts as an accessible and valuable source of natural products with potential wound-healing activity, supporting the sustainable use of these underexploited agricultural materials.