Effects of amygdalin on ferroptosis and oxidative stress in diabetic retinopathy progression via the NRF2/ARE signaling pathway.

ABSTRACT

Oxidative stress has been involved in the pathogenesis of diabetic retinopathy (DR). Amygdalin is an effective component of bitter almond that exhibits excellent antioxidant properties. We explored the effects of amygdalin on ferroptosis and oxidative stress in high-glucose (HG)-stimulated human retinal endothelial cells (HRECs) via the NRF2/ARE pathway. HG-stimulated HRECs were used to establish a DR model. Cell viability was evaluated using the MTT assay. The release of lactate dehydrogenase was used to evaluate cell toxicity. The protein levels of NRF2, NQO1, and HO-1 were detected using western blotting. The GSH, GSSG, GPX4, SOD, CAT, MDA, and Fe2+ levels in the HRECs were also detected. Flow cytometry was used to detect reactive oxygen species (ROS) using a fluorescent probe. Immunofluorescence staining was performed to detect NRF2 expression. The results revealed that HG stimulation decreased the levels of GSH, GPX4, SOD, and CAT but increased those of MDA, ROS, GSSG, and Fe2+ in HRECs. Ferrostatin-1 treatment reversed the effects of HG stimulation, whereas erastin aggravated these effects. Amygdalin treatment relieved HG-induced injury in HRECs. Amygdalin treatment promoted the nuclear transport of NRF2 in HG-stimulated HRECs. NQO1 and HO-1 levels were upregulated in HG-stimulated HRECs after amygdalin treatment. An inhibitor of NRF2 reversed the effects of amygdalin. Therefore, amygdalin treatment inhibited ferroptosis and oxidative stress in HG-stimulated HRECs by activating the NRF2/ARE signaling pathway.