Ophthalmology Department, Zhongshan Hospital Affiliated to Xiamen University, No.201-209 Hubinnan Road, Siming District, 361004, Xiamen, China.
Endocrinology Department, Beijing Electric Power Hospital, 100073, Beijing, China.
BMC Ophthalmol. 2023 Sep 6;23(1):368. doi: 10.1186/s12886-023-03115-9.
Maresin-1 plays an important role in diabetic illnesses and ferroptosis is associated with pathogenic processes of diabetic retinopathy (DR). The goal of this study is to explore the influence of maresin-1 on ferroptosis and its molecular mechanism in DR.
ARPE-19 cells were exposed to high glucose (HG) condition for developing a cellular model of DR. The CCK-8 assay and flow cytometry were used to assess ARPE-19 cell proliferation and apoptosis, respectively. Furthermore, the GSH content, MDA content, ROS level, and Fe level were measured by using a colorimetric GSH test kit, a Lipid Peroxidation MDA Assay Kit, a DCFH-DA assay and the phirozine technique, respectively. Immunofluorescence labelling was used to detect protein levels of ACSL4 and PTGS2. Messenger RNA and protein expression of HO-1, GPX4 and Nrf2 was evaluated through western blotting and quantitative real time-polymerase chain reaction (qRT-PCR). To establish a diabetic mouse model, mice were intraperitoneally injected 150 mg/kg streptozotocin. The MDA content, ROS level and the iron level were detected by using corresponding commercial kits.
Maresin-1 promoted cell proliferation while reducing the apoptotic process in HG-induced ARPE-19 cells. Maresin-1 significantly reduced ferroptosis induced by HG in ARPE-19 cells, as demonstrated as a result of decreased MDA content, ROS level, Fe level, PTGS2 expression, ACSL4 expression and increased GSH content. With respect to mechanisms, maresin-1 treatment up-regulated the mRNA expression and protein expression of HO-1, GPX4 and Nrf2 in HG-induced ARPE-19 cells. Nrf2 inhibitor reversed the inhibitory effects of maresin-1 on ferroptosis in HG-induced ARPE-19 cells. In vivo experiments, we found that Maresin-1 evidently repressed ferroptosis a mouse model of DR, as evidenced by the decreased MDA content, ROS level and iron level in retinal tissues of mice.
Maresin-1 protects ARPE cells from HG-induced ferroptosis via activating the Nrf2/HO-1/GPX4 pathway, suggesting that maresin-1 prevents DR development.
maresin-1 在糖尿病疾病中发挥重要作用，铁死亡与糖尿病性视网膜病变 (DR) 的发病过程有关。本研究旨在探讨maresin-1 对 DR 中铁死亡的影响及其分子机制。
用高糖 (HG) 处理 ARPE-19 细胞，建立 DR 细胞模型。用 CCK-8 法和流式细胞术分别检测 ARPE-19 细胞增殖和凋亡情况。此外，用比色法 GSH 试剂盒、脂质过氧化 MDA 测定试剂盒、DCFH-DA 法和菲咯嗪法分别测定细胞内 GSH 含量、MDA 含量、ROS 水平和 Fe 水平。用免疫荧光标记法检测 ACSL4 和 PTGS2 蛋白水平。用 Western blot 和 qRT-PCR 法检测 HO-1、GPX4 和 Nrf2 的信使 RNA 和蛋白表达。用腹腔注射 150mg/kg 链脲佐菌素建立糖尿病小鼠模型。用相应的商业试剂盒检测 MDA 含量、ROS 水平和铁水平。
maresin-1 促进 HG 诱导的 ARPE-19 细胞增殖，减少细胞凋亡。maresin-1 显著降低 HG 诱导的 ARPE-19 细胞铁死亡，表现为 MDA 含量、ROS 水平、Fe 水平、PTGS2 表达、ACSL4 表达降低，GSH 含量增加。机制上，maresin-1 处理可上调 HG 诱导的 ARPE-19 细胞中 HO-1、GPX4 和 Nrf2 的 mRNA 表达和蛋白表达。Nrf2 抑制剂逆转了 maresin-1 对 HG 诱导的 ARPE-19 细胞中铁死亡的抑制作用。在体内实验中，我们发现 Maresin-1 可明显抑制 DR 小鼠模型中的铁死亡，表现为视网膜组织中 MDA 含量、ROS 水平和铁水平降低。
maresin-1 通过激活 Nrf2/HO-1/GPX4 通路保护 ARPE 细胞免受 HG 诱导的铁死亡，提示 maresin-1 可预防 DR 的发生。
