Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression
Abstracts Background Many neurological diseases involve neuroinflammation, during which overproduction of cytokines by immune cells, especially microglia, can aggregate neuronal death. Ferroptosis is a recently discovered cell metabolism-related form of cell death and RSL3 is a well-known inducer of...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-10-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12974-021-02231-x |
| _version_ | 1818732532175208448 |
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| author | Yu Cui Zhaolong Zhang Xin Zhou Zhiyuan Zhao Rui Zhao Xiangyu Xu Xiangyi Kong Jinyang Ren Xujin Yao Qian Wen Feifei Guo Shengli Gao Jiangdong Sun Qi Wan |
| author_facet | Yu Cui Zhaolong Zhang Xin Zhou Zhiyuan Zhao Rui Zhao Xiangyu Xu Xiangyi Kong Jinyang Ren Xujin Yao Qian Wen Feifei Guo Shengli Gao Jiangdong Sun Qi Wan |
| author_sort | Yu Cui |
| collection | DOAJ |
| description | Abstracts Background Many neurological diseases involve neuroinflammation, during which overproduction of cytokines by immune cells, especially microglia, can aggregate neuronal death. Ferroptosis is a recently discovered cell metabolism-related form of cell death and RSL3 is a well-known inducer of cell ferroptosis. Here, we aimed to investigate the effects of RSL3 in neuroinflammation and sensitivity of different type of microglia and macrophage to ferroptosis. Methods Here, we used quantitative RT-PCR analysis and ELISA analysis to analyze the production of proinflammatory cytokine production of microglia and macrophages after lipopolysaccharides (LPS) stimulation. We used CCK8, LDH, and flow cytometry analysis to evaluate the sensitivity of different microglia and macrophages to RSL3-induced ferroptosis. Western blot was used to test the activation of inflammatory signaling pathway and knockdown efficiency. SiRNA-mediated interference was conducted to knockdown GPX4 or Nrf2 in BV2 microglia. Intraperitoneal injection of LPS was performed to evaluate systemic inflammation and neuroinflammation severity in in vivo conditions. Results We found that ferroptosis inducer RSL3 inhibited lipopolysaccharides (LPS)-induced inflammation of microglia and peritoneal macrophages (PMs) in a cell ferroptosis-independent manner, whereas cell ferroptosis-conditioned medium significantly triggered inflammation of microglia and PMs. Different type of microglia and macrophages showed varied sensitivity to RSL3-induced ferroptosis. Mechanistically, RSL3 induced Nrf2 protein expression to inhibit RNA Polymerase II recruitment to transcription start site of proinflammatory cytokine genes to repress cytokine transcription, and protect cells from ferroptosis. Furthermore, simultaneously injection of RSL3 and Fer-1 ameliorated LPS-induced neuroinflammation in in vivo conditions. Conclusions These data revealed the proinflammatory role of ferroptosis in microglia and macrophages, identified RSL3 as a novel inhibitor of LPS-induced inflammation, and uncovered the molecular regulation of microglia and macrophage sensitivity to ferroptosis. Thus, targeting ferroptosis in diseases by using RSL3 should consider both the pro-ferroptosis effect and the anti-inflammation effect to achieve optimal outcome. |
| first_indexed | 2024-12-17T23:35:04Z |
| format | Article |
| id | doaj.art-a3d79428444946f09e810a57eaceb5a0 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-12-17T23:35:04Z |
| publishDate | 2021-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-a3d79428444946f09e810a57eaceb5a02022-12-21T21:28:35ZengBMCJournal of Neuroinflammation1742-20942021-10-0118111510.1186/s12974-021-02231-xMicroglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expressionYu Cui0Zhaolong Zhang1Xin Zhou2Zhiyuan Zhao3Rui Zhao4Xiangyu Xu5Xiangyi Kong6Jinyang Ren7Xujin Yao8Qian Wen9Feifei Guo10Shengli Gao11Jiangdong Sun12Qi Wan13Institute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityThe Affiliated Hospital of Qingdao UniversityInstitute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityThe Affiliated Hospital of Qingdao UniversityThe Affiliated Hospital of Qingdao UniversityThe Affiliated Hospital of Qingdao UniversityInstitute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityInstitute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityInstitute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityDepartment of Biomedical Center, Qingdao UniversitySchool of Basic Medicine, Qingdao UniversitySchool of Basic Medicine, Qingdao UniversityInstitute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityInstitute of Neuroregeneration & Neurorehabilitation, Department of Pathophysiology, Qingdao UniversityAbstracts Background Many neurological diseases involve neuroinflammation, during which overproduction of cytokines by immune cells, especially microglia, can aggregate neuronal death. Ferroptosis is a recently discovered cell metabolism-related form of cell death and RSL3 is a well-known inducer of cell ferroptosis. Here, we aimed to investigate the effects of RSL3 in neuroinflammation and sensitivity of different type of microglia and macrophage to ferroptosis. Methods Here, we used quantitative RT-PCR analysis and ELISA analysis to analyze the production of proinflammatory cytokine production of microglia and macrophages after lipopolysaccharides (LPS) stimulation. We used CCK8, LDH, and flow cytometry analysis to evaluate the sensitivity of different microglia and macrophages to RSL3-induced ferroptosis. Western blot was used to test the activation of inflammatory signaling pathway and knockdown efficiency. SiRNA-mediated interference was conducted to knockdown GPX4 or Nrf2 in BV2 microglia. Intraperitoneal injection of LPS was performed to evaluate systemic inflammation and neuroinflammation severity in in vivo conditions. Results We found that ferroptosis inducer RSL3 inhibited lipopolysaccharides (LPS)-induced inflammation of microglia and peritoneal macrophages (PMs) in a cell ferroptosis-independent manner, whereas cell ferroptosis-conditioned medium significantly triggered inflammation of microglia and PMs. Different type of microglia and macrophages showed varied sensitivity to RSL3-induced ferroptosis. Mechanistically, RSL3 induced Nrf2 protein expression to inhibit RNA Polymerase II recruitment to transcription start site of proinflammatory cytokine genes to repress cytokine transcription, and protect cells from ferroptosis. Furthermore, simultaneously injection of RSL3 and Fer-1 ameliorated LPS-induced neuroinflammation in in vivo conditions. Conclusions These data revealed the proinflammatory role of ferroptosis in microglia and macrophages, identified RSL3 as a novel inhibitor of LPS-induced inflammation, and uncovered the molecular regulation of microglia and macrophage sensitivity to ferroptosis. Thus, targeting ferroptosis in diseases by using RSL3 should consider both the pro-ferroptosis effect and the anti-inflammation effect to achieve optimal outcome.https://doi.org/10.1186/s12974-021-02231-xFerroptosisRSL3NeuroinflammationMicrogliaMacrophageLPS |
| spellingShingle | Yu Cui Zhaolong Zhang Xin Zhou Zhiyuan Zhao Rui Zhao Xiangyu Xu Xiangyi Kong Jinyang Ren Xujin Yao Qian Wen Feifei Guo Shengli Gao Jiangdong Sun Qi Wan Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression Journal of Neuroinflammation Ferroptosis RSL3 Neuroinflammation Microglia Macrophage LPS |
| title | Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression |
| title_full | Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression |
| title_fullStr | Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression |
| title_full_unstemmed | Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression |
| title_short | Microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after RSL3 stimulation via increasing Nrf2 expression |
| title_sort | microglia and macrophage exhibit attenuated inflammatory response and ferroptosis resistance after rsl3 stimulation via increasing nrf2 expression |
| topic | Ferroptosis RSL3 Neuroinflammation Microglia Macrophage LPS |
| url | https://doi.org/10.1186/s12974-021-02231-x |
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