Pancreatic β-cells are highly susceptible to ferroptosis, an iron-dependent form of cell death driven by lipid peroxidation, which contributes to β-cell dysfunction in Type 2 diabetes mellitus (T2DM). This study investigated the anti-ferroptotic potential of Niruryadi Gulika (NG), a traditional polyherbal formulation, using an integrative network pharmacology and in vitro approach. Bioactive compounds of NG were identified from databases, and their potential protein targets were predicted. Ferroptosis-associated genes were compiled, and overlapping targets were analyzed through protein–protein interaction (PPI) networks, gene ontology (GO), and KEGG pathway enrichment analyses. Experimental validation was conducted in MIN6 pancreatic β-cells exposed to high-glucose and cytokine-induced (HGM) stress. Cell viability, gene expression (GPx4, CAT, SOD2, p22phox, TXNIP, TRPC6), and protein levels of key ferroptosis markers (GPx4, ACSL4, SOD2) were assessed. Network analysis identified 22 NG compounds targeting 452 proteins, with 90 overlapping ferroptosis-associated genes; GPx4 emerged as a central hub gene. Enrichment analysis revealed the involvement of oxidative stress and cell death pathways. NG (10 and 50 µg/mL) significantly improved β-cell viability under HGM stress, upregulated antioxidant genes (GPx4, CAT, SOD2), and downregulated oxidative stress markers (p22phox, TXNIP, TRPC6). Western blot analysis confirmed increased GPx4 and SOD2 protein expression alongside suppression of the pro-ferroptotic protein ACSL4. Collectively, these findings demonstrate that NG exerts cytoprotective effects against ferroptosis in pancreatic β-cells by enhancing antioxidant defence and inhibiting lipid peroxidation pathways, supporting its potential as a multi-target therapeutic candidate for T2DM.
Kumar et al. (Fri,) studied this question.