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

Abstract

Abstract 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.