Ferrostatin-1 alleviates tissue and cell damage in diabetic retinopathy by improving the antioxidant capacity of the Xc--GPX4 system

Abstract

Diabetic retinopathy (DR) is a common microvascular complication leading to a high blindness rate among patients with diabetes. Ferroptosis is a type of cell death caused by the accumulation of iron-dependent lipid peroxides. Studies have shown that ferroptosis plays an important role in DR. The rat model of DR was constructed and treated with Ferrostatin-1 (Ferr-1). Haematoxylin and eosin (HE) were used to detect the degree of retinopathy. Oxidative stress levels were detected by ELISA. Perl’s staining was used to detect iron deposition in retinal tissues. Ferritin levels were measured by ELISA. The expression of GPX4 was detected by immunohistochemistry (IHC). GSH/GSSG kit was used to detect the content and proportion of reduced/oxidized glutathione. Western blot was used to detect the expression of ferroptosis-related proteins. TUNEL assay was used to detect cell apoptosis. The expression of GSDMD was detected by fluorescence in situ hybridization (FISH). Western blot was used to detect the expression of apoptosis and pyroptosis-related proteins. Then, high glucose (HG)-induced retinal epithelial cell line ARPE-19 was treated by Erastin (ferroptosis activator) and Ferr-1. CCK-8, ELISA, western blot, flow cytometry, and immunofluorescence (IF) staining were used to detect oxidative stress levels, ferroptosis and cell damage. The mechanism was further explored by adding ferroptosis agonist Erastin. In vitro and in vivo results showed that oxidative stress was increased in DR model, resulting in ferroptosis and tissue or cell damage. After administration of Ferr-1, the antioxidant capacity was improved, ferroptosis levels were reduced and tissue or cell damage was alleviated. In vitro results showed that Ferr-1 reversed the impacts of Erastin on oxidative stress, ferroptosis, and cell damage in HG-induced ARPE-19 cells. Ferr-1 alleviated tissue and cell damage by improving the antioxidant capacity of the Xc--GPX4 system.

Mesenchymal Stem Cell-derived Exosomal miR-125b-5p Suppressed Retinal Microvascular Endothelial Cell Ferroptosis by Targeting P53 in Diabetic Retinopathy

“…However, the underlying mechanisms are still unclear. Ferroptosis has been reported to contribute to DR [21]. In the present study, we examined ferroptosis in STZ-induced DR and con rmed that MSC-sEVs could protect vascular endothelial function and maintain BRB stability.…”

Arsenic trioxide induces ferroptosis in neuroblastoma by mediating GPX4 transcriptional inhibition

“…Giovanni Miotto et al demonstrated that Fer-1 exerted an inhibitory effect on ferroptosis by scavenging the initial intracellular alkoxy radicals and elevating GSH levels. 42,43 The process of intracellular GSH against ferroptosis required the involvement of the rate-limiting enzyme GPX4. In the ATOtreated NB cells, the transcription of GPX4 was markedly suppressed, leading to a notable reduction in the protein expression level of GPX4.…”

“…Giovanni Miotto et al. demonstrated that Fer‐1 exerted an inhibitory effect on ferroptosis by scavenging the initial intracellular alkoxy radicals and elevating GSH levels 42,43 . The process of intracellular GSH against ferroptosis required the involvement of the rate‐limiting enzyme GPX4.…”

Design, synthesis, and evaluation of novel ferrostatin derivatives for the prevention of HG-induced VEC ferroptosis

“…19 It has demonstrated cytoprotective effects against ferroptosis induced by different stimuli, including erastin, RSL3, and HG. 17,20 However, the low potency, poor solubility, and stability impose on the clinic application of Fer-1 which is a great limitation. To overcome these shortcomings, a few improved analogs of Fer-1, such as SRS9-11, SRS9-92, SRS9-86, and UMAC-2148 (Fig.…”