Acteoside alleviates high glucose-induced ROS accumulation and ferroptosis in ARPE-19 cells by activating the NRF2 signaling pathway, and upregulates NRF2 to inhibit ferroptosis in the retina of diabetic retinopathy mice, thereby ameliorating retinal injury.
Key Findings
Results
High glucose conditions significantly reduced ARPE-19 cell viability and promoted ferroptosis markers compared to normal glucose conditions.
ARPE-19 cells were induced with 33 mmol/L high glucose (HG) to construct an in vitro diabetic retinopathy model.
HG group showed significantly reduced cell viability compared to normal glucose (NG) group.
HG increased intracellular Fe2+ levels and lipid peroxidation.
HG decreased glutathione (GSH) content and downregulated expression of GPX4 and SLC7A11 proteins.
Results
Ferroptosis inhibitors Ferrostatin-1 (Fer-1) and deferoxamine (DFO) significantly ameliorated HG-induced reductions in cell viability and increases in lipid peroxidation.
Fer-1 and DFO are established ferroptosis inhibitors used to confirm ferroptosis involvement.
Both inhibitors significantly ameliorated HG-induced reductions in cell viability.
Both inhibitors significantly reduced HG-induced increases in lipid peroxidation.
These results confirmed that HG-induced cell damage in ARPE-19 cells involves ferroptosis.
Results
Acteoside (ACT) intervention markedly reversed HG-induced promotion of ferroptosis in ARPE-19 cells.
ACT treatment reversed HG-induced changes in ferroptosis markers including Fe2+ levels, lipid peroxidation, and GSH content.
ACT upregulated expression of GPX4 and SLC7A11, key anti-ferroptosis proteins.
Ferroptosis was assessed using CCK-8 assay, measurement kits, BODIPY-C11 staining, and Western blot.
ACT also reduced ROS accumulation in HG-treated ARPE-19 cells.
Results
ACT significantly promoted NRF2 nuclear translocation, and silencing NRF2 weakened ACT's inhibitory effect on ferroptosis.
NRF2 nuclear translocation was examined by immunofluorescence.
NRF2 silencing experiments demonstrated that NRF2 is required for ACT's anti-ferroptotic effects.
These results indicate ACT's protective effects are mediated through the NRF2 signaling pathway.
Results
In a streptozotocin (STZ)-induced mouse diabetic retinopathy model, ACT intervention significantly alleviated retinal tissue damage in a dose-dependent manner.
A C57BL/6 mouse DR model was established via intraperitoneal injection of STZ and treated with different doses of ACT.
ACT intervention groups exhibited significantly alleviated retinal tissue damage as assessed by HE staining.
ACT-treated mice showed reduced tissue Fe2+, 4-HNE, and MDA levels, and increased GSH content.
ACT upregulated expression of NRF2, GPX4, and SLC7A11 proteins in retinal tissue.
Effects showed a dose-dependent trend across ACT treatment groups.
What This Means
This research suggests that a natural compound called Acteoside (ACT), found in certain plants, may help protect the retina from damage caused by high blood sugar, as seen in diabetic retinopathy (DR). The researchers tested this in retinal pigment epithelial cells (ARPE-19) exposed to high glucose levels in the lab and in diabetic mice. They found that high glucose triggers a specific form of cell death called ferroptosis — a process involving iron accumulation and oxidative damage to fats in cells — and that ACT could block this process. ACT worked by activating a protective cellular pathway controlled by a protein called NRF2, which in turn boosted the production of protective molecules like GPX4 and SLC7A11 that defend against ferroptosis.
In mouse experiments, diabetic mice treated with ACT showed less retinal damage, lower levels of harmful oxidative stress markers (such as Fe2+, 4-HNE, and MDA), higher levels of the protective molecule glutathione (GSH), and increased expression of NRF2 and its downstream proteins. These benefits became more pronounced at higher doses of ACT, suggesting a dose-dependent effect. When researchers artificially silenced the NRF2 gene in cells, ACT's protective effects were weakened, confirming that NRF2 activation is central to how ACT works.
This research suggests that Acteoside could be a promising candidate for further study as a treatment to slow or prevent retinal damage in diabetic retinopathy by targeting the ferroptosis pathway. However, these findings are based on laboratory cell cultures and animal models, and further research — including clinical trials in humans — would be needed to determine whether ACT is safe and effective for people with diabetic eye disease.
Shen S, Chen Q, Li Y, Ma J, Zhang Y, Luo H, et al.. (2026). Acteoside alleviates high glucose-induced ROS accumulation and ferroptosis in ARPE-19 cells by activating the NRF2 signaling pathway.. Molecular biology reports. https://doi.org/10.1007/s11033-026-12608-x