Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance
Objective To investigate the role of transcription factor EB (TFEB) in endotoxin-tolerant macrophages. Methods The RAW264.7 cells were divided into blank group (DMEM medium), LPS 5 group (5 ng/mL LPS treatment for 4 h), LPS 100 group (100 ng/mL LPS treatment for 4 h), and tolerance group (5 ng/mL...
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Editorial Office of Journal of Army Medical University
2025-04-01
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| Series: | 陆军军医大学学报 |
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| Online Access: | https://aammt.tmmu.edu.cn/html/202410042.html |
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| author | YANG Ting YANG Ting LIU Xin |
| author_facet | YANG Ting YANG Ting LIU Xin |
| author_sort | YANG Ting |
| collection | DOAJ |
| description | Objective To investigate the role of transcription factor EB (TFEB) in endotoxin-tolerant macrophages. Methods The RAW264.7 cells were divided into blank group (DMEM medium), LPS 5 group (5 ng/mL LPS treatment for 4 h), LPS 100 group (100 ng/mL LPS treatment for 4 h), and tolerance group (5 ng/mL LPS for 12 h followed by 100 ng/mL LPS for 4 h). The releases of inflammatory factors TNF-α and IL-6 were measured using ELISA. Western blotting and immunofluorescence assay were used to evaluate the distribution of autophagy-related proteins LC3 and P62, as well as TFEB in the cytoplasm and nucleus. Lentiviral overexpression of TFEB or siRNA-mediated knockdown of TFEB were performed to observe the changes in autophagy levels and bacterial clearance ability in the tolerant cells. Results The cells in the tolerance group had significantly lower contents of TNF-α and IL-6, as well as reduced bacterial clearance ability (P<0.01), down-regulated LC3 expression while up-regulated P62 level, and decreased expression of TFEB in both the cytoplasm and nucleus (P<0.01) when compared with the cells of the LPS 100 group. Overexpression of TFEB significantly increased LC3 level, reduced P62 level, and enhanced bacterial clearance ability in the endotoxin-tolerant cells (P<0.01). In contrast, siRNA-mediated knockdown of TFEB had no significant impacts on LC3 and P62 expression levels or bacterial clearance ability. Conclusion Overexpression of TFEB can restore the autophagy of endotoxin-tolerant cells and enhance their bacterial clearance capacity, thereby alleviating the immunosuppressive state of sepsis. These findings suggest that TFEB holds promise as a potential therapeutic target for the prevention and treatment of sepsis.
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| format | Article |
| id | doaj-art-7ad5f6c36f174e8ebb1a929d8fd83f32 |
| institution | DOAJ |
| issn | 2097-0927 |
| language | zho |
| publishDate | 2025-04-01 |
| publisher | Editorial Office of Journal of Army Medical University |
| record_format | Article |
| series | 陆军军医大学学报 |
| spelling | doaj-art-7ad5f6c36f174e8ebb1a929d8fd83f322025-08-20T03:14:19ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272025-04-0147879480610.16016/j.2097-0927.202410042Transcription factor EB enhances macrophage autophagy and reverses endotoxin toleranceYANG Ting0YANG Ting1LIU Xin2Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology， Faculty of Pharmacology， College of Pharmacy， Chongqing Medical University， ChongqingClinical Research Center， First Affiliated Hospital， Army Medical University （Third Military Medical University）， Chongqing， ChinaClinical Research Center， First Affiliated Hospital， Army Medical University （Third Military Medical University）， Chongqing， ChinaObjective To investigate the role of transcription factor EB (TFEB) in endotoxin-tolerant macrophages. Methods The RAW264.7 cells were divided into blank group (DMEM medium), LPS 5 group (5 ng/mL LPS treatment for 4 h), LPS 100 group (100 ng/mL LPS treatment for 4 h), and tolerance group (5 ng/mL LPS for 12 h followed by 100 ng/mL LPS for 4 h). The releases of inflammatory factors TNF-α and IL-6 were measured using ELISA. Western blotting and immunofluorescence assay were used to evaluate the distribution of autophagy-related proteins LC3 and P62, as well as TFEB in the cytoplasm and nucleus. Lentiviral overexpression of TFEB or siRNA-mediated knockdown of TFEB were performed to observe the changes in autophagy levels and bacterial clearance ability in the tolerant cells. Results The cells in the tolerance group had significantly lower contents of TNF-α and IL-6, as well as reduced bacterial clearance ability (P<0.01), down-regulated LC3 expression while up-regulated P62 level, and decreased expression of TFEB in both the cytoplasm and nucleus (P<0.01) when compared with the cells of the LPS 100 group. Overexpression of TFEB significantly increased LC3 level, reduced P62 level, and enhanced bacterial clearance ability in the endotoxin-tolerant cells (P<0.01). In contrast, siRNA-mediated knockdown of TFEB had no significant impacts on LC3 and P62 expression levels or bacterial clearance ability. Conclusion Overexpression of TFEB can restore the autophagy of endotoxin-tolerant cells and enhance their bacterial clearance capacity, thereby alleviating the immunosuppressive state of sepsis. These findings suggest that TFEB holds promise as a potential therapeutic target for the prevention and treatment of sepsis. https://aammt.tmmu.edu.cn/html/202410042.htmlsepsisimmunosuppressionendotoxin tolerancetranscription factor ebautophagy |
| spellingShingle | YANG Ting YANG Ting LIU Xin Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance 陆军军医大学学报 sepsis immunosuppression endotoxin tolerance transcription factor eb autophagy |
| title | Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance |
| title_full | Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance |
| title_fullStr | Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance |
| title_full_unstemmed | Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance |
| title_short | Transcription factor EB enhances macrophage autophagy and reverses endotoxin tolerance |
| title_sort | transcription factor eb enhances macrophage autophagy and reverses endotoxin tolerance |
| topic | sepsis immunosuppression endotoxin tolerance transcription factor eb autophagy |
| url | https://aammt.tmmu.edu.cn/html/202410042.html |
| work_keys_str_mv | AT yangting transcriptionfactorebenhancesmacrophageautophagyandreversesendotoxintolerance AT yangting transcriptionfactorebenhancesmacrophageautophagyandreversesendotoxintolerance AT liuxin transcriptionfactorebenhancesmacrophageautophagyandreversesendotoxintolerance |