O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway
Abstract Background Lung ischemia–reperfusion (I/R) injury is a common clinical pathology associated with high mortality. The pathophysiology of lung I/R injury involves ferroptosis and elevated protein O-GlcNAcylation levels, while the effect of O-GlcNAcylation on lung I/R injury remains unclear. T...
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2025-02-01
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| Online Access: | https://doi.org/10.1186/s12915-025-02126-w |
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| author | Liuqing Yang Hexiao Tang Jin Wang Dawei Xu Rui Xuan Songping Xie Pengfei Xu Xinyi Li |
| author_facet | Liuqing Yang Hexiao Tang Jin Wang Dawei Xu Rui Xuan Songping Xie Pengfei Xu Xinyi Li |
| author_sort | Liuqing Yang |
| collection | DOAJ |
| description | Abstract Background Lung ischemia–reperfusion (I/R) injury is a common clinical pathology associated with high mortality. The pathophysiology of lung I/R injury involves ferroptosis and elevated protein O-GlcNAcylation levels, while the effect of O-GlcNAcylation on lung I/R injury remains unclear. This research aimed to explore the effect of O-GlcNAcylation on reducing ferroptosis in pulmonary epithelial cells caused by I/R. Results First, we identified O-GlcNAc transferase 1 (Ogt1) as a differentially expressed gene in lung epithelial cells of acute lung injury/acute respiratory distress syndrome (ALI/ARDS) patients, using single-cell sequencing, and Gene Ontology analysis (GO analysis) revealed the enrichment of the ferroptosis process. We found a time-dependent dynamic alteration in lung O-GlcNAcylation during I/R injury. Proteomics analysis identified the differentially expressed proteins enriched in ferroptosis and multiple redox-related pathways based on KEGG annotation. Thus, we generated Ogt1-conditional knockout mice and found that Ogt1 deficiency aggravated ferroptosis, as evidenced by lipid reactive oxygen species (lipid ROS), malondialdehyde (MDA), Fe2+, as well as alterations in critical protein expression glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11). Consistently, we found that elevated O-GlcNAcylation inhibited ferroptosis sensitivity in hypoxia/reoxygenation (H/R) injury-induced TC-1 cells via O-GlcNAcylated NF-E2-related factor-2 (Nrf2). Furthermore, both the chromatin immunoprecipitation (ChIP) assay and the dual-luciferase reporter assay indicated that Nrf2 could bind with translation start site (TSS) of glucose-6-phosphate dehydrogenase (G6PDH) and promote its transcriptional activity. As an important rate-limiting enzyme in the pentose phosphate pathway (PPP), elevated G6PDH provided a mass of nicotinamide adenine dinucleotide phosphate (NADPH) to improve the redox state of glutathione (GSH) and eventually led to ferroptosis resistance. Rescue experiments proved that Nrf2 knockdown or Nrf2-T334A (O-GlcNAcylation site) mutation abolished the protective effect of ferroptosis resistance. Conclusions In summary, we revealed that O-GlcNAcylation could protect against I/R lung injury by reducing ferroptosis sensitivity via the Nrf2/G6PDH pathway. Our work will provide a new basis for clinical therapeutic strategies for pulmonary ischemia–reperfusion-induced acute lung injury. |
| format | Article |
| id | doaj-art-93f6b23e0e494e14a00893391a9c1524 |
| institution | Kabale University |
| issn | 1741-7007 |
| language | English |
| publishDate | 2025-02-01 |
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| spelling | doaj-art-93f6b23e0e494e14a00893391a9c15242025-02-09T12:54:24ZengBMCBMC Biology1741-70072025-02-0123112310.1186/s12915-025-02126-wO-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathwayLiuqing Yang0Hexiao Tang1Jin Wang2Dawei Xu3Rui Xuan4Songping Xie5Pengfei Xu6Xinyi Li7Department of Anesthesiology, Zhongnan Hospital of Wuhan UniversityDepartment of Thoracic Surgery, Zhongnan Hospital of Wuhan UniversityDepartment of Anesthesiology, Zhongnan Hospital of Wuhan UniversityDepartment of Anesthesiology, Zhongnan Hospital of Wuhan UniversityDepartment of Anesthesiology, Zhongnan Hospital of Wuhan UniversityDepartment of Thoracic Surgery, Renmin Hospital of Wuhan UniversityDepartment of Hepatobiliary and Pancreatic Surgery, School of Pharmaceutical Sciences, Zhongnan Hospital of Wuhan University, Wuhan UniversityDepartment of Anesthesiology, Zhongnan Hospital of Wuhan UniversityAbstract Background Lung ischemia–reperfusion (I/R) injury is a common clinical pathology associated with high mortality. The pathophysiology of lung I/R injury involves ferroptosis and elevated protein O-GlcNAcylation levels, while the effect of O-GlcNAcylation on lung I/R injury remains unclear. This research aimed to explore the effect of O-GlcNAcylation on reducing ferroptosis in pulmonary epithelial cells caused by I/R. Results First, we identified O-GlcNAc transferase 1 (Ogt1) as a differentially expressed gene in lung epithelial cells of acute lung injury/acute respiratory distress syndrome (ALI/ARDS) patients, using single-cell sequencing, and Gene Ontology analysis (GO analysis) revealed the enrichment of the ferroptosis process. We found a time-dependent dynamic alteration in lung O-GlcNAcylation during I/R injury. Proteomics analysis identified the differentially expressed proteins enriched in ferroptosis and multiple redox-related pathways based on KEGG annotation. Thus, we generated Ogt1-conditional knockout mice and found that Ogt1 deficiency aggravated ferroptosis, as evidenced by lipid reactive oxygen species (lipid ROS), malondialdehyde (MDA), Fe2+, as well as alterations in critical protein expression glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11). Consistently, we found that elevated O-GlcNAcylation inhibited ferroptosis sensitivity in hypoxia/reoxygenation (H/R) injury-induced TC-1 cells via O-GlcNAcylated NF-E2-related factor-2 (Nrf2). Furthermore, both the chromatin immunoprecipitation (ChIP) assay and the dual-luciferase reporter assay indicated that Nrf2 could bind with translation start site (TSS) of glucose-6-phosphate dehydrogenase (G6PDH) and promote its transcriptional activity. As an important rate-limiting enzyme in the pentose phosphate pathway (PPP), elevated G6PDH provided a mass of nicotinamide adenine dinucleotide phosphate (NADPH) to improve the redox state of glutathione (GSH) and eventually led to ferroptosis resistance. Rescue experiments proved that Nrf2 knockdown or Nrf2-T334A (O-GlcNAcylation site) mutation abolished the protective effect of ferroptosis resistance. Conclusions In summary, we revealed that O-GlcNAcylation could protect against I/R lung injury by reducing ferroptosis sensitivity via the Nrf2/G6PDH pathway. Our work will provide a new basis for clinical therapeutic strategies for pulmonary ischemia–reperfusion-induced acute lung injury.https://doi.org/10.1186/s12915-025-02126-wFerroptosisG6PDHLung ischemia–reperfusionNrf2Ogt1O-GlcNAcylation |
| spellingShingle | Liuqing Yang Hexiao Tang Jin Wang Dawei Xu Rui Xuan Songping Xie Pengfei Xu Xinyi Li O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway BMC Biology Ferroptosis G6PDH Lung ischemia–reperfusion Nrf2 Ogt1 O-GlcNAcylation |
| title | O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway |
| title_full | O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway |
| title_fullStr | O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway |
| title_full_unstemmed | O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway |
| title_short | O-GlcNAcylation attenuates ischemia-reperfusion-induced pulmonary epithelial cell ferroptosis via the Nrf2/G6PDH pathway |
| title_sort | o glcnacylation attenuates ischemia reperfusion induced pulmonary epithelial cell ferroptosis via the nrf2 g6pdh pathway |
| topic | Ferroptosis G6PDH Lung ischemia–reperfusion Nrf2 Ogt1 O-GlcNAcylation |
| url | https://doi.org/10.1186/s12915-025-02126-w |
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