Wang Yi, Song Si-Yuan, Song Yi, Wang Yuan, Wan Zheng-Wei, Sun Ping, Yu Xue-Mei, Deng Bo, Zeng Kai-Hong
Clinical Immunology Translational Medicine Key Laboratory of Sichuan Province, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, China.
Baylor College of Medicine, Houston, TX, 77030, USA.
Mol Neurobiol. 2025 Mar;62(3):3412-3427. doi: 10.1007/s12035-024-04496-8. Epub 2024 Sep 18.
The aim of this study was to investigate the anti-ferroptotic effect of resveratrol (RSV) on retinal Müller cells (RMCs) in the early stages of diabetic retinopathy (DR) via the nuclear factor erythroid 2-related factor 2 (Nrf2)/glutathione peroxidase 4 (GPx4)/prostaglandin-endoperoxide synthase 2 (PTGS2). The retina was obtained from normal and diabetic Sprague-Dawley rats or wild-type and Nrf2 knockout (KO) diabetic mice, with or without RSV (10 mg/kg/d) treatment for 12 weeks. RMCs transfected with or without SiNrf2 were cultured with high glucose and RSV (20 mM). The retinal neurofunctional changes were measured by electroretinogram (ERG). The retinal inner nuclear layer cell mitochondrial morphological changes were detected by transmission electron microscopy. The cell viabilities were measured by cell counting kit-8 (CCK-8) assay. The levels of Fe, malonic dialdehyde (MDA), and glutathione (GSH) were measured by colorimetric method. The expression of Nrf2, GPx4, and PTGS2 was detected by quantitative real-time polymerase chain reaction (qRT-PCR), western blotting, and immunocytochemistry. In vivo, RSV inhibited retinal neurofunctional changes and mitochondrial morphological changes; decreased Fe, MDA, and PTGS2; and increased GSH, Nrf2, and GPx4 in retina of DM rats. In vitro, RSV decreased MDA and PTGS2 and increased cell viability, GSH, Nrf2, and GPx4. In vivo and vitro, the role of Nrf2-regulated signaling pathway in anti-ferroptosis by RSV was further confirmed using Nrf2 KO mice and pre-transfected SiNrf2 in RMCs. These findings indicated that RSV is a potential therapeutic option for DR and that Nrf2/GPx4/PTGS2 plays a role in the anti-ferroptosis mechanism of RSV on RMCs.
本研究旨在通过核因子红细胞2相关因子2（Nrf2）/谷胱甘肽过氧化物酶4（GPx4）/前列腺素内过氧化物合酶2（PTGS2），探讨白藜芦醇（RSV）对糖尿病视网膜病变（DR）早期视网膜Müller细胞（RMCs）的抗铁死亡作用。从正常和糖尿病的Sprague-Dawley大鼠或野生型和Nrf2基因敲除（KO）的糖尿病小鼠获取视网膜，分别给予或不给予RSV（10 mg/kg/d）治疗12周。用或不用SiNrf2转染的RMCs在高糖和RSV（20 mM）条件下培养。通过视网膜电图（ERG）测量视网膜神经功能变化。用透射电子显微镜检测视网膜内核层细胞线粒体形态变化。用细胞计数试剂盒-8（CCK-8）法测量细胞活力。用比色法测量铁、丙二醛（MDA）和谷胱甘肽（GSH）水平。通过定量实时聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法和免疫细胞化学检测Nrf2、GPx4和PTGS2的表达。在体内，RSV抑制糖尿病大鼠视网膜神经功能变化和线粒体形态变化；降低铁、MDA和PTGS2水平；增加GSH、Nrf2和GPx4水平。在体外，RSV降低MDA和PTGS2水平并提高细胞活力、GSH、Nrf2和GPx4水平。在体内和体外，使用Nrf2基因敲除小鼠和预先在RMCs中转染SiNrf2进一步证实了Nrf2调节的信号通路在RSV抗铁死亡中的作用。这些研究结果表明，RSV是DR的一种潜在治疗选择，且Nrf2/GPx4/PTGS2在RSV对RMCs的抗铁死亡机制中发挥作用。
