Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways
Background: The expression of Na-K-2Cl cotransporter 1 (NKCC1) in the alveolar epithelium is responsible for fluid homeostasis in acute lung injury (ALI). Increasing evidence suggests that NKCC1 is associated with inflammation in ALI. We hypothesized that inhibiting NKCC1 would attenuate ALI after i...
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Frontiers Media S.A.
2018-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2018.02049/full |
| _version_ | 1819067529813819392 |
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| author | Chih-Hao Shen Chih-Hao Shen Chih-Hao Shen Jr-Yu Lin Yung-Lung Chang Shu-Yu Wu Chung-Kan Peng Chin-Pyng Wu Kun-Lun Huang Kun-Lun Huang |
| author_facet | Chih-Hao Shen Chih-Hao Shen Chih-Hao Shen Jr-Yu Lin Yung-Lung Chang Shu-Yu Wu Chung-Kan Peng Chin-Pyng Wu Kun-Lun Huang Kun-Lun Huang |
| author_sort | Chih-Hao Shen |
| collection | DOAJ |
| description | Background: The expression of Na-K-2Cl cotransporter 1 (NKCC1) in the alveolar epithelium is responsible for fluid homeostasis in acute lung injury (ALI). Increasing evidence suggests that NKCC1 is associated with inflammation in ALI. We hypothesized that inhibiting NKCC1 would attenuate ALI after ischemia-reperfusion (IR) by modulating pathways that are mediated by tumor necrosis-associated factor 6 (TRAF6).Methods: IR-ALI was induced by producing 30 min of ischemia followed by 90 min of reperfusion in situ in an isolated and perfused rat lung model. The rats were randomly allotted into four groups comprising two control groups and two IR groups with and without bumetanide. Alveolar fluid clearance (AFC) was measured for each group. Mouse alveolar MLE-12 cells were cultured in control and hypoxia-reoxygenation (HR) conditions with or without bumetanide. Flow cytometry and transwell monolayer permeability assay were carried out for each group.Results: Bumetanide attenuated the activation of p-NKCC1 and lung edema after IR. In the HR model, bumetanide decreased the cellular volume and increased the transwell permeability. In contrast, bumetanide increased the expression of epithelial sodium channel (ENaC) via p38 mitogen-activated protein kinase (p38 MAPK), which attenuated the reduction of AFC after IR. Bumetanide also modulated lung inflammation via nuclear factor-κB (NF-κB). TRAF6, which is upstream of p38 MAPK and NF-κB, was attenuated by bumetanide after IR and HR.Conclusions: Inhibition of NKCC1 by bumetanide reciprocally modulated epithelial p38 MAPK and NF-κB via TRAF6 in IR-ALI. This interaction attenuated the reduction of AFC via upregulating ENaC expression and reduced lung inflammation. |
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| id | doaj.art-1102d8f8c58c4c73ab87ae4ddcbb3625 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-21T16:19:42Z |
| publishDate | 2018-09-01 |
| publisher | Frontiers Media S.A. |
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| spelling | doaj.art-1102d8f8c58c4c73ab87ae4ddcbb36252022-12-21T18:57:36ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-09-01910.3389/fimmu.2018.02049396195Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated PathwaysChih-Hao Shen0Chih-Hao Shen1Chih-Hao Shen2Jr-Yu Lin3Yung-Lung Chang4Shu-Yu Wu5Chung-Kan Peng6Chin-Pyng Wu7Kun-Lun Huang8Kun-Lun Huang9Division of Pulmonary and Critical Care, Department of Internal Medicine, Tri-Service General Hospital, Taipei, TaiwanGraduate Institute of Medical Sciences, National Defense Medical Center, Taipei, TaiwanInstitute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, TaiwanInstitute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, TaiwanDepartment of Biochemistry, National Defense Medical Center, Taipei, TaiwanInstitute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, TaiwanDivision of Pulmonary and Critical Care, Department of Internal Medicine, Tri-Service General Hospital, Taipei, TaiwanDepartment of Critical Care Medicine, Landseed Hospital, Taoyuan, TaiwanDivision of Pulmonary and Critical Care, Department of Internal Medicine, Tri-Service General Hospital, Taipei, TaiwanInstitute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, TaiwanBackground: The expression of Na-K-2Cl cotransporter 1 (NKCC1) in the alveolar epithelium is responsible for fluid homeostasis in acute lung injury (ALI). Increasing evidence suggests that NKCC1 is associated with inflammation in ALI. We hypothesized that inhibiting NKCC1 would attenuate ALI after ischemia-reperfusion (IR) by modulating pathways that are mediated by tumor necrosis-associated factor 6 (TRAF6).Methods: IR-ALI was induced by producing 30 min of ischemia followed by 90 min of reperfusion in situ in an isolated and perfused rat lung model. The rats were randomly allotted into four groups comprising two control groups and two IR groups with and without bumetanide. Alveolar fluid clearance (AFC) was measured for each group. Mouse alveolar MLE-12 cells were cultured in control and hypoxia-reoxygenation (HR) conditions with or without bumetanide. Flow cytometry and transwell monolayer permeability assay were carried out for each group.Results: Bumetanide attenuated the activation of p-NKCC1 and lung edema after IR. In the HR model, bumetanide decreased the cellular volume and increased the transwell permeability. In contrast, bumetanide increased the expression of epithelial sodium channel (ENaC) via p38 mitogen-activated protein kinase (p38 MAPK), which attenuated the reduction of AFC after IR. Bumetanide also modulated lung inflammation via nuclear factor-κB (NF-κB). TRAF6, which is upstream of p38 MAPK and NF-κB, was attenuated by bumetanide after IR and HR.Conclusions: Inhibition of NKCC1 by bumetanide reciprocally modulated epithelial p38 MAPK and NF-κB via TRAF6 in IR-ALI. This interaction attenuated the reduction of AFC via upregulating ENaC expression and reduced lung inflammation.https://www.frontiersin.org/article/10.3389/fimmu.2018.02049/fullNa-K-2Cl cotransporter 1bumetanideischemia-reperfusionacute lung injuryepithelial sodium channelp38 mitogen-activated protein kinase |
| spellingShingle | Chih-Hao Shen Chih-Hao Shen Chih-Hao Shen Jr-Yu Lin Yung-Lung Chang Shu-Yu Wu Chung-Kan Peng Chin-Pyng Wu Kun-Lun Huang Kun-Lun Huang Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways Frontiers in Immunology Na-K-2Cl cotransporter 1 bumetanide ischemia-reperfusion acute lung injury epithelial sodium channel p38 mitogen-activated protein kinase |
| title | Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways |
| title_full | Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways |
| title_fullStr | Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways |
| title_full_unstemmed | Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways |
| title_short | Inhibition of NKCC1 Modulates Alveolar Fluid Clearance and Inflammation in Ischemia-Reperfusion Lung Injury via TRAF6-Mediated Pathways |
| title_sort | inhibition of nkcc1 modulates alveolar fluid clearance and inflammation in ischemia reperfusion lung injury via traf6 mediated pathways |
| topic | Na-K-2Cl cotransporter 1 bumetanide ischemia-reperfusion acute lung injury epithelial sodium channel p38 mitogen-activated protein kinase |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2018.02049/full |
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