Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis
BackgroundNotch signaling is critical for regulating the function of vascular endothelial cells (ECs). However, the effect of the intracellular domain of Notch1 (NICD) on EC injury in sepsis remains unclear.MethodsWe established a cell model of vascular endothelial dysfunction and induced sepsis in...
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| Format: | Article |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1134556/full |
| _version_ | 1827956009601073152 |
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| author | Tingyan Liu Caiyan Zhang Jiayun Ying Yaodong Wang Gangfeng Yan Yufeng Zhou Yufeng Zhou Guoping Lu |
| author_facet | Tingyan Liu Caiyan Zhang Jiayun Ying Yaodong Wang Gangfeng Yan Yufeng Zhou Yufeng Zhou Guoping Lu |
| author_sort | Tingyan Liu |
| collection | DOAJ |
| description | BackgroundNotch signaling is critical for regulating the function of vascular endothelial cells (ECs). However, the effect of the intracellular domain of Notch1 (NICD) on EC injury in sepsis remains unclear.MethodsWe established a cell model of vascular endothelial dysfunction and induced sepsis in a mouse model via lipopolysaccharide (LPS) injection and cecal ligation and puncture (CLP). Endothelial barrier function and expression of endothelial-related proteins were determined using CCK-8, permeability, flow cytometry, immunoblot, and immunoprecipitation assays. The effect of NICD inhibition or activation on endothelial barrier function was evaluated in vitro. Melatonin was used for NICD activation in sepsis mice. The survival rate, Evans blue dye of organs, vessel relaxation assay, immunohistochemistry, ELISA, immunoblot were used to explore the specific role of melatonin for sepsis induced vascular dysfunction in vivo.ResultsWe found that LPS, interleukin 6, and serum collected from septic children could inhibit the expression of NICD and its downstream regulator Hes1, which impaired endothelial barrier function and led to EC apoptosis through the AKT pathway. Mechanistically, LPS decreased the stability of NICD by inhibiting the expression of a deubiquitylating enzyme, ubiquitin-specific proteases 8 (USP8). Melatonin, however, upregulated USP8 expression, thus maintaining the stability of NICD and Notch signaling, which ultimately reduced EC injury in our sepsis model and elevated the survival rate of septic mice.ConclusionsWe found a previously uncharacterized role of Notch1 in mediating vascular permeability during sepsis, and we showed that inhibition of NICD resulted in vascular EC dysfunction in sepsis, which was reversed by melatonin. Thus, the Notch1 signaling pathway is a potential target for the treatment of sepsis. |
| first_indexed | 2024-04-09T14:56:51Z |
| format | Article |
| id | doaj.art-d7ba1d58066348cbb806069d8afcdc52 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-09T14:56:51Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-d7ba1d58066348cbb806069d8afcdc522023-05-02T05:09:19ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-05-011410.3389/fimmu.2023.11345561134556Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsisTingyan Liu0Caiyan Zhang1Jiayun Ying2Yaodong Wang3Gangfeng Yan4Yufeng Zhou5Yufeng Zhou6Guoping Lu7Department of Critical Care Medicine, Children’s Hospital of Fudan University, Shanghai, ChinaDepartment of Critical Care Medicine, Children’s Hospital of Fudan University, Shanghai, ChinaDepartment of Critical Care Medicine, Children’s Hospital of Fudan University, Shanghai, ChinaDepartment of Critical Care Medicine, Children’s Hospital of Fudan University, Shanghai, ChinaDepartment of Critical Care Medicine, Children’s Hospital of Fudan University, Shanghai, ChinaInstitute of Pediatrics, Children’s Hospital of Fudan University, National Children’s Medical Center, and the Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Institutes of Biomedical Sciences, Fudan University, Shanghai, ChinaNational Health Commission (NHC) Key Laboratory of Neonatal Diseases, Fudan University, Shanghai, ChinaDepartment of Critical Care Medicine, Children’s Hospital of Fudan University, Shanghai, ChinaBackgroundNotch signaling is critical for regulating the function of vascular endothelial cells (ECs). However, the effect of the intracellular domain of Notch1 (NICD) on EC injury in sepsis remains unclear.MethodsWe established a cell model of vascular endothelial dysfunction and induced sepsis in a mouse model via lipopolysaccharide (LPS) injection and cecal ligation and puncture (CLP). Endothelial barrier function and expression of endothelial-related proteins were determined using CCK-8, permeability, flow cytometry, immunoblot, and immunoprecipitation assays. The effect of NICD inhibition or activation on endothelial barrier function was evaluated in vitro. Melatonin was used for NICD activation in sepsis mice. The survival rate, Evans blue dye of organs, vessel relaxation assay, immunohistochemistry, ELISA, immunoblot were used to explore the specific role of melatonin for sepsis induced vascular dysfunction in vivo.ResultsWe found that LPS, interleukin 6, and serum collected from septic children could inhibit the expression of NICD and its downstream regulator Hes1, which impaired endothelial barrier function and led to EC apoptosis through the AKT pathway. Mechanistically, LPS decreased the stability of NICD by inhibiting the expression of a deubiquitylating enzyme, ubiquitin-specific proteases 8 (USP8). Melatonin, however, upregulated USP8 expression, thus maintaining the stability of NICD and Notch signaling, which ultimately reduced EC injury in our sepsis model and elevated the survival rate of septic mice.ConclusionsWe found a previously uncharacterized role of Notch1 in mediating vascular permeability during sepsis, and we showed that inhibition of NICD resulted in vascular EC dysfunction in sepsis, which was reversed by melatonin. Thus, the Notch1 signaling pathway is a potential target for the treatment of sepsis.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1134556/fullNICDvascular endothelial dysfunctionmelatoninLPS (lipopolysaccharide)sepsis |
| spellingShingle | Tingyan Liu Caiyan Zhang Jiayun Ying Yaodong Wang Gangfeng Yan Yufeng Zhou Yufeng Zhou Guoping Lu Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis Frontiers in Immunology NICD vascular endothelial dysfunction melatonin LPS (lipopolysaccharide) sepsis |
| title | Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis |
| title_full | Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis |
| title_fullStr | Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis |
| title_full_unstemmed | Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis |
| title_short | Inhibition of the intracellular domain of Notch1 results in vascular endothelial cell dysfunction in sepsis |
| title_sort | inhibition of the intracellular domain of notch1 results in vascular endothelial cell dysfunction in sepsis |
| topic | NICD vascular endothelial dysfunction melatonin LPS (lipopolysaccharide) sepsis |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1134556/full |
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