Type 2 diabetes mellitus (T2DM) progression is closely associated with oxidative stress–mediated cellular damage. Ferroptosis is a regulated, iron-dependent form of cell death driven by lipid peroxidation and an imbalance between reactive oxygen species (ROS) and antioxidant defenses. Puerarin, a naturally occurring isoflavonoid, has documented antioxidant properties, but its capacity to ameliorate T2DM via modulation of ferroptosis had not been clarified prior to this investigation.
The authors combined network pharmacology and proteomics to predict and identify potential molecular targets of puerarin in T2DM. These bioinformatic and proteomic findings were then tested experimentally. In vitro experiments used a palmitic acid model to induce oxidative stress in mouse pancreatic β-cells (MIN6). Molecular and biochemical assays and Western blot analysis were applied to evaluate oxidative stress markers, ferroptosis-related proteins, and pathway intermediates. Functional perturbations included AKT1 overexpression and knockdown models and application of nuclear factor erythroid 2–related factor 2 (NRF2) inhibitors to probe pathway dependencies.
To evaluate ferroptosis and oxidative stress, the study measured:
Ferroptosis-specific and related protein markers measured included:
Insulin levels were also assessed as a functional readout relevant to pancreatic β-cell biology.
In MIN6 mouse pancreatic β-cells, palmitic acid–induced oxidative stress provoked features consistent with ferroptosis. Under puerarin treatment, multiple indices shifted in a direction consistent with ferroptosis suppression: alterations in ROS, MDA, Mn-SOD, intracellular iron, and GSH were reported, alongside changes in mitochondrial morphology. The authors report that these changes reflect mitigation of oxidative damage and reduced ferroptotic activity in the cell model following puerarin intervention.
Mechanistic interrogation used AKT1 overexpression and knockdown models together with NRF2 inhibitors and Western blotting. The reported chain of effects is:
Taken together, these results indicate that puerarin may reduce ferroptosis by activating an AKT/GSK3β/NRF2/GPX4 signaling axis, thereby enhancing antioxidant capacity and GPX4-mediated protection against lipid peroxidation.
The integrated network pharmacology and proteomics approach identified potential targets of puerarin in the context of T2DM; AKT1 emerged as a key target in these analyses. Experimental validation focused on AKT1 because of its central role in the observed signaling changes and functional outcomes related to ferroptosis suppression.
This study presents combined bioinformatic, proteomic, and experimental evidence that puerarin can mitigate oxidative stress–induced ferroptosis in a pancreatic β-cell model of T2DM. The proposed mechanism involves puerarin-mediated upregulation of AKT1, promotion of GSK3β phosphorylation, increased NRF2 expression, and subsequent elevation of GPX4, which supports glutathione metabolism and counters ferroptotic lipid peroxidation. The authors describe this integrated identification of puerarin targets and the link to the AKT/GSK3β/NRF2/GPX4 pathway as a novel mechanistic insight with potential relevance for diabetes treatment strategies.
The abstract reports in vitro palmitic acid–induced MIN6 cell experiments, network pharmacology and proteomics analyses, AKT1 modulation models, use of NRF2 inhibitors, and Western blot confirmation of pathway effects. The abstract does not report in vivo data, clinical data, specific dosing regimens, long-term outcomes, or safety/tolerability information. Details on exact proteomics results, statistical metrics, and experimental protocols beyond those summarized were not provided in the abstract of the source article.