An integrative evaluation of in vitro anti-inflammatory potential, network pharmacology and molecular docking of Girardinia diversifolia (Urticaceae) relevant to rheumatoid arthritis.

Abstract

Girardinia diversifolia is a wild medicinal plant traditionally used to treat rheumatism. However, experimental validation of its anti-inflammatory potential remained unexplored. This study investigated the prospect of G. diversifolia leaf extracts to alleviate inflammation via in vitro and in silico approach with relevance to rheumatoid arthritis (RA). Phytochemical composition of methanolic extract of G. diversifolia leaves (GD) were investigated via spectrophotometric methods, gas chromatography - mass spectrometry (GC-MS) and High-Resolution Liquid Chromatography-Mass Spectrometry Quadrupole Time-of-Flight (HR-LCMS-QROF). In vitro anti-inflammatory activity was evaluated via HRBC membrane stabilization assay, heat induced protein denaturation assay and 15-lipoxygenase inhibitory assay. Additional anti-oxidant assays were conducted. Network pharmacology analysis was conducted to predict potential RA associated gene targets of detected GD compounds, hub genes and signalling pathways. Molecular docking of compounds to target hub genes and evaluation of interacting residues was done. Phytochemical analysis tentatively identified several phytosterols, fatty acids, tocopherols, alkaloids and terpenoids. At the concentrations tested, GD exhibited notable membrane stabilizing activity, prevention of protein denaturation and 15-LOX inhibitory activity. At higher concentrations membrane stabilization and prevention of protein denaturation was statistically comparable to standard drug. The predicted bioactive compounds - Squalene, Somniferine, Americine and Dipiperamide E showed favourable binding affinities for key inflammatory targets - TLR4, AKT1, MMP9 and EGFR respectively, interacting with key residues present at the canonical binding sites of the proteins. In conclusion, G. diversifolia has anti-inflammatory potential. The present findings provide preliminary scientific evidence for utilization of G. diversifolia as traditional medicine for treating rheumatism and establish a foundation for future pharmacological investigations of this underexplored medicinal species.

Supplementary information: The online version contains supplementary material available at https://doi.org/10.1007/s40203-026-00730-4.