Department of Ophthalmology, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, No. 358 Datong Road, Pudong New Area, Shanghai, 200137, China.
Department of Ophthalmolog, Shanghai Eye Diseases Prevention & Treatment Center/Shanghai Eye Hospital, No.380 Kangding Road, Shanghai, 200041, China.
Exp Eye Res. 2023 Sep;234:109569. doi: 10.1016/j.exer.2023.109569. Epub 2023 Jul 6.
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 Fe 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 Fe 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.
氧化应激与糖尿病性视网膜病变（DR）的发病机制有关。苦杏仁苷是苦杏仁的有效成分，具有出色的抗氧化特性。我们通过 NRF2/ARE 通路探讨了苦杏仁苷对高糖（HG）刺激的人视网膜内皮细胞（HRECs）中铁死亡和氧化应激的影响。使用 HG 刺激的 HRECs 建立 DR 模型。使用 MTT 测定法评估细胞活力。通过测定乳酸脱氢酶的释放来评估细胞毒性。使用 Western blot 检测 NRF2、NQO1 和 HO-1 的蛋白水平。还检测了 HRECs 中的 GSH、GSSG、GPX4、SOD、CAT、MDA 和 Fe 水平。使用荧光探针通过流式细胞术检测活性氧（ROS）。通过免疫荧光染色检测 NRF2 表达。结果表明，HG 刺激降低了 GSH、GPX4、SOD 和 CAT 的水平，但增加了 MDA、ROS、GSSG 和 Fe 的水平。铁死亡抑制剂 1（Ferrostatin-1）处理逆转了 HG 刺激的作用，而 erastin 则加重了这些作用。苦杏仁苷处理缓解了 HG 诱导的 HRECs 损伤。苦杏仁苷处理促进了 HG 刺激的 HRECs 中 NRF2 的核转运。在 HG 刺激的 HRECs 中，经苦杏仁苷处理后 NQO1 和 HO-1 水平上调。NRF2 抑制剂逆转了苦杏仁苷的作用。因此，苦杏仁苷通过激活 NRF2/ARE 信号通路抑制 HG 刺激的 HRECs 中的铁死亡和氧化应激。
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