Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis
AIM: To investigate the protective role of ghrelin against diabetic retinopathy (DR), focusing on its anti-ferroptotic mechanism in high glucose-induced retinal endothelial injury. METHODS: First, small interfering RNA (siRNA)-mediated interference was conducted to knockdown nuclear factor erythroid...
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Press of International Journal of Ophthalmology (IJO PRESS)
2025-06-01
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| Online Access: | http://ies.ijo.cn/en_publish/2025/6/20250602.pdf |
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| author | Fen-Fen Wang Rong Li Dan Liao Chao-Qun Liu Xiao-Li Yang |
| author_facet | Fen-Fen Wang Rong Li Dan Liao Chao-Qun Liu Xiao-Li Yang |
| author_sort | Fen-Fen Wang |
| collection | DOAJ |
| description | AIM: To investigate the protective role of ghrelin against diabetic retinopathy (DR), focusing on its anti-ferroptotic mechanism in high glucose-induced retinal endothelial injury. METHODS: First, small interfering RNA (siRNA)-mediated interference was conducted to knockdown nuclear factor erythroid 2-related factor 2 (Nrf2). Using reverse transcription-polymerase chain reaction (RT-PCR), the expression level of Nrf2 was determined from human retinal microvascular endothelial cells (HRMECs) transfected with either si-NC or si-Nrf2. After that, cells were treated with 10 nmol/L ghrelin and then cultured in a high glucose (30 mmol/L) environment. EdU assay was utilized to assess cell proliferation, while transmission electron microscopy was employed to observe mitochondrial morphology. Flow cytometry was used to measure the level of intracellular reactive oxygen species (ROS), and biochemical assays were conducted to detect malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), and ferrous iron (Fe2+). Western blotting was used to identify the presence of ferroptosis-related proteins such as glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), Nrf2, and haem oxygenase-1 (HO-1). RESULTS: Under a high glucose environment, ghrelin could significantly promote the proliferation of HRMECs and mitochondrial status, remarkably decrease the levels of intracellular ROS and MDA, and up-regulate the level of GSH and SOD. Besides, ghrelin greatly reduced Fe2+ level in the cells while increased protein levels of GPX4 and SLC7A11. Subsequently, we found that high glucose induced inactivation of Nrf2/HO-1 axis and the protein expression profile were significantly promoted by ghrelin. Moreover, silencing of Nrf2 by siRNA delivery markedly diminished the changes induced by ghrelin in high glucose-induced HRMECs, shown as reduced cell proliferation and increased mitochondrial malformation, up-regulated ROS, MDA, Fe2+, GPX4 and SLC7A11, as well as down-regulated GSH, SOD, Nrf2 and HO-1. CONCLUSION: Ghrelin attenuates high glucose-induced injury of retinal endothelial cells via inhibiting ferroptosis, and activation of Nrf2/HO-1 pathway may be one of the mechanisms involved in this effect of ghrelin. |
| format | Article |
| id | doaj-art-58b20784c3d64705bfcd2db40dababb5 |
| institution | DOAJ |
| issn | 2222-3959 2227-4898 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Press of International Journal of Ophthalmology (IJO PRESS) |
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| series | International Journal of Ophthalmology |
| spelling | doaj-art-58b20784c3d64705bfcd2db40dababb52025-08-20T03:13:03ZengPress of International Journal of Ophthalmology (IJO PRESS)International Journal of Ophthalmology2222-39592227-48982025-06-0118697898510.18240/ijo.2025.06.0220250602Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosisFen-Fen Wang0Rong Li1Dan Liao2Chao-Qun Liu3Xiao-Li Yang4Rong Li. Department of Ophthalmology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China. rechelrong198222@163.comDepartment of Ophthalmology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Medical School of Ophthalmology and Optometry, North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Department of Ophthalmology, the First Affiliated Hospital of Xi'an Medical University, Xi'an 710077, Shaanxi Province, ChinaDepartment of Ophthalmology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Medical School of Ophthalmology and Optometry, North Sichuan Medical College, Nanchong 637000, Sichuan Province, ChinaDepartment of Ophthalmology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Medical School of Ophthalmology and Optometry, North Sichuan Medical College, Nanchong 637000, Sichuan Province, ChinaDepartment of Ophthalmology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Medical School of Ophthalmology and Optometry, North Sichuan Medical College, Nanchong 637000, Sichuan Province, ChinaAIM: To investigate the protective role of ghrelin against diabetic retinopathy (DR), focusing on its anti-ferroptotic mechanism in high glucose-induced retinal endothelial injury. METHODS: First, small interfering RNA (siRNA)-mediated interference was conducted to knockdown nuclear factor erythroid 2-related factor 2 (Nrf2). Using reverse transcription-polymerase chain reaction (RT-PCR), the expression level of Nrf2 was determined from human retinal microvascular endothelial cells (HRMECs) transfected with either si-NC or si-Nrf2. After that, cells were treated with 10 nmol/L ghrelin and then cultured in a high glucose (30 mmol/L) environment. EdU assay was utilized to assess cell proliferation, while transmission electron microscopy was employed to observe mitochondrial morphology. Flow cytometry was used to measure the level of intracellular reactive oxygen species (ROS), and biochemical assays were conducted to detect malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), and ferrous iron (Fe2+). Western blotting was used to identify the presence of ferroptosis-related proteins such as glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), Nrf2, and haem oxygenase-1 (HO-1). RESULTS: Under a high glucose environment, ghrelin could significantly promote the proliferation of HRMECs and mitochondrial status, remarkably decrease the levels of intracellular ROS and MDA, and up-regulate the level of GSH and SOD. Besides, ghrelin greatly reduced Fe2+ level in the cells while increased protein levels of GPX4 and SLC7A11. Subsequently, we found that high glucose induced inactivation of Nrf2/HO-1 axis and the protein expression profile were significantly promoted by ghrelin. Moreover, silencing of Nrf2 by siRNA delivery markedly diminished the changes induced by ghrelin in high glucose-induced HRMECs, shown as reduced cell proliferation and increased mitochondrial malformation, up-regulated ROS, MDA, Fe2+, GPX4 and SLC7A11, as well as down-regulated GSH, SOD, Nrf2 and HO-1. CONCLUSION: Ghrelin attenuates high glucose-induced injury of retinal endothelial cells via inhibiting ferroptosis, and activation of Nrf2/HO-1 pathway may be one of the mechanisms involved in this effect of ghrelin.http://ies.ijo.cn/en_publish/2025/6/20250602.pdfghrelinhuman retinal microvascular endothelial cellsferroptosisnuclear factor erythroid 2-related factor 2haem oxygenase-1oxidative stress |
| spellingShingle | Fen-Fen Wang Rong Li Dan Liao Chao-Qun Liu Xiao-Li Yang Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis International Journal of Ophthalmology ghrelin human retinal microvascular endothelial cells ferroptosis nuclear factor erythroid 2-related factor 2 haem oxygenase-1 oxidative stress |
| title | Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis |
| title_full | Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis |
| title_fullStr | Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis |
| title_full_unstemmed | Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis |
| title_short | Ghrelin alleviates high glucose-induced retinal microvascular endothelial cell injury by activating Nrf2/HO-1 pathway to inhibit ferroptosis |
| title_sort | ghrelin alleviates high glucose induced retinal microvascular endothelial cell injury by activating nrf2 ho 1 pathway to inhibit ferroptosis |
| topic | ghrelin human retinal microvascular endothelial cells ferroptosis nuclear factor erythroid 2-related factor 2 haem oxygenase-1 oxidative stress |
| url | http://ies.ijo.cn/en_publish/2025/6/20250602.pdf |
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