Tumor necrosis factor-α requires Ezrin to regulate the cytoskeleton and cause pulmonary microvascular endothelial barrier damage
| العنوان: | Tumor necrosis factor-α requires Ezrin to regulate the cytoskeleton and cause pulmonary microvascular endothelial barrier damage |
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| المؤلفون: | Gengyun Sun, Juan Sun, Na Zhang, Jianjun Jiang, Sihui Tang |
| المصدر: | Microvascular Research |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Male, rac1 GTP-Binding Protein, ARDS, RAC1, macromolecular substances, Ezrin, 030204 cardiovascular system & hematology, Biochemistry, Article, Permeability, Small hairpin RNA, Pathogenesis, Capillary Permeability, Rats, Sprague-Dawley, 03 medical and health sciences, 0302 clinical medicine, Western blot, medicine, PMVEC, Electric Impedance, Animals, Phosphorylation, Cytoskeleton, Lung, Cells, Cultured, medicine.diagnostic_test, Chemistry, Tumor Necrosis Factor-alpha, Endothelial Cells, Cell Biology, medicine.disease, Cell biology, Cytoskeletal Proteins, 030104 developmental biology, Microvessels, Tumor necrosis factor alpha, Cardiology and Cardiovascular Medicine, Rac1 |
| الوصف: | Acute respiratory distress syndrome (ARDS) is a rapidly progressive disease with unknown pathogenesis. Damage of pulmonary microvascular endothelial cells(PMVECs) caused by inflammatory storm caused by cytokines such as TNF- α is the potential pathogenesis of ARDS.In this study, we examined the role of ezrin and Rac1 in TNF-α-related pathways, which regulates the permeability of PMVECs. Primary rat pulmonary microvascular endothelial cells (RPMVECs) were isolated and cultured. RPMVECs were treated with rat TNF - α (0,1,10,100ng / ml), and the cell activity of each group was measured using a CCK8 kit. The integrity of endothelial barrier was measured by transendothelial resistance (TEER) and FITC-BSA flux across RPMVECs membranes. Pulldown assay and WesternBlot was used to detect the activity of RAS-associated C3 botulinum toxin substrate 1 (Rac1) and Ezrin phosphorylation. Short hairpin RNA(shRNA) targeting ezrin and Rac1 was utilized to evaluate the effectof RPMVECs permeability and related pathway. The effects of ezrin and Rac1 on cytoskeleton were confirmed by immunofluorescence.Our results revealed that active Rac1 was essential for protecting the RPMVEC barrier stimulated by TNF-α, while active ezrin could partially destroy the PMVEC barrier by reducing Rac1 activity and regulating the subcellular structure of the cytoskeleton. These findings may be used to create new therapeutic strategies for targeting Rac1 in the treatment of ARDS. Highlights • Ezrin silencing alleviates the damage of the PMVEC barrier caused by TNF- α. • Ezrin silencing partially reversed TNF-α induced cytoskeleton rearrangements and cortactin redistribution in PMVEC. • Ezrin silencing increased the expression of active Rac1 in PMVECs. • Rac1 silencing induced a significant increase in ezrin phosphorylation in PMVECs. |
| تدمد: | 1095-9319 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ef5732771703a567c6d181235913c0fa https://pubmed.ncbi.nlm.nih.gov/33007316 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....ef5732771703a567c6d181235913c0fa |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10959319 |
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