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Endothelial mitochondria determine rapid barrier failure in chemical lung injury.

ABSTRACT: Acid aspiration, which can result from several etiologies, including postoperative complications, leads to direct contact of concentrated hydrochloric acid (HCl) with the alveolar epithelium. As a result, rapid endothelial activation induces alveolar inflammation, leading to life-threatening pulmonary edema. Because mechanisms underlying the rapid endothelial activation are not understood, here we determined responses in real time through optical imaging of alveoli of live mouse lungs. By alveolar micropuncture, we microinfused concentrated HCl in the alveolar lumen. As expected, acid contact with the epithelium caused rapid, but transient, apical injury. However, there was no concomitant membrane injury to the endothelium. Nevertheless, H2O2-mediated epithelial-endothelial paracrine signaling induced endothelial barrier failure, as detected by microvascular dextran leakage and lung water quantification. Remarkably, endothelial mitochondria regulated the barrier failure by activating uncoupling protein 2 (UCP2), thereby inducing transient mitochondrial depolarization that led to cofilin-induced actin depolymerization. Knockdown, or endothelium-targeted deletion of UCP2 expression, blocked these responses, including pulmonary edema. To our knowledge, these findings are the first to mechanistically implicate endothelial mitochondria in acid-induced barrier deterioration and pulmonary edema. We suggest endothelial UCP2 may be a therapeutic target for acid-induced acute lung injury.

SUBMITTER: Hough RF

PROVIDER: S-EPMC6413830 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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Endothelial mitochondria determine rapid barrier failure in chemical lung injury.

Hough Rebecca F RF Islam Mohammad N MN Gusarova Galina A GA Jin Guangchun G Das Shonit S Bhattacharya Jahar J

JCI insight 20190207 3

Acid aspiration, which can result from several etiologies, including postoperative complications, leads to direct contact of concentrated hydrochloric acid (HCl) with the alveolar epithelium. As a result, rapid endothelial activation induces alveolar inflammation, leading to life-threatening pulmonary edema. Because mechanisms underlying the rapid endothelial activation are not understood, here we determined responses in real time through optical imaging of alveoli of live mouse lungs. By alveol ...[more]

PMID: 30728333

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Project description:BACKGROUND:Endothelial barrier dysfunction is central to the pathogenesis of sepsis-associated acute lung injury (ALI). Microtubule (MT) dynamics in vascular endothelium are crucial for the regulation of endothelial barrier function. Unfractionated heparin (UFH) possesses various biological activities, such as anti-inflammatory activity and endothelial barrier protection during sepsis. METHODS:Here, we investigated the effects and underlying mechanisms of UFH on lipopolysaccharide (LPS)-induced endothelial barrier dysfunction. C57BL/6 J mice were randomized into vehicle, UFH, LPS and LPS?+?UFH groups. Intraperitoneal injection of 30 mg/kg LPS was used to induce sepsis. Mice in the LPS?+?UFH group received intravenous UFH 0.5 h prior to LPS injection. Human pulmonary microvascular endothelial cells (HPMECs) were cultured for analyzing the effects of UFH on LPS-induced and nocodazole-induced hyperpermeability, F-actin remodeling, and LPS-induced p38 MAPK activation. RESULTS:UFH pretreatment significantly attenuated LPS-induced pulmonary histopathological changes, and increased the lung W/D ratio and Evans blue accumulation in vivo. Both in vivo and in vitro studies showed that UFH pretreatment blocked the LPS-induced increase in guanine nucleotide exchange factor (GEF-H1) expression and myosin phosphatase target subunit 1 (MYPT1) phosphorylation, and microtubule (MT) disassembly in LPS-induced ALI mouse model and human pulmonary microvascular endothelial cells (HPMECs). These results suggested that UFH ameliorated LPS-induced endothelial barrier dysfunction by inhibiting MT disassembly and GEF-H1 expression. In addition, UFH attenuated LPS-induced hyperpermeability of HPMECs and F-actin remodeling. In vitro, UFH pretreatment inhibited LPS-induced increase in monomeric tubulin expression and decrease in tubulin polymerization and acetylation. Meanwhile, UFH ameliorates nocodazole-induced MTs disassembly and endothelial barrier dysfunction.Additionally, UFH decreased p38 phosphorylation and activation, which was similar to the effect of the p38 MAPK inhibitor, SB203580. CONCLUSIONS:UFH exert its protective effects on pulmonary microvascular endothelial barrier dysfunction via microtubule stabilization and is associated with the p38 MAPK pathway.

Project description:BackgroundAcute lung injury (ALI)/acute respiratory distress syndrome (ARDS) has long posed challenges in clinical practice, lacking established preventive and therapeutic approaches. Lianhua Qingke (LHQK), a patented traditional Chinese medicine (TCM), has been found to have anti-inflammatory effects for ameliorating ALI/ARDS induced by lipopolysaccharide (LPS). This study aimed to investigate the effects and potential mechanisms of LHQK on endothelial protection in LPS-induced ALI/ARDS in vivo and in LPS-induced human pulmonary microvascular endothelial cells (HPMECs) injury in vitro.MethodsIn the animal experiment, we induced an ALI/ARDS model by intratracheal injection of LPS (5 mg/mL). LHQK (3.7 g/kg/d for low dose and 7.4 g/kg/d for high dose) or dexamethasone (DEX) (5 mg/kg/d) was administered to mice 3 days prior to LPS treatment. In the in vitro experiments, HPMECs were pretreated with LHQK at concentrations of 125 and 250 µg/mL for 2 hours before being stimulated with LPS (10 µg/mL). We employed lung function test, measurement of lung index, hematoxylin and eosin (H&E) staining, bronchoalveolar lavage fluid (BALF) cell counts, and inflammatory cytokine levels to assess the therapeutic effect of LHQK. Additionally, the extravasation assay of fluorescein isothiocyanate-dextran (FITC-dextran) dye and the transmembrane electrical resistance (TEER) assay were used to evaluate endothelial barrier. Barrier integrity and relevant protein validation were assessed using immunofluorescence (IF) and Western blot analyses. Furthermore, network pharmacology analysis and cellular level screening were employed to predict and screen the active ingredients of LHQK.ResultsCompared to the LPS group, LHQK significantly improved lung function, mitigated lung pathological injuries, reduced inflammatory cells and inflammatory cytokines [tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6] levels in BALF, and inhibited the expression of vascular cell adhesion molecule-1 (VCAM-1), attenuated LPS-induced pulmonary oedema and FITC-dextran permeability, and enhanced the expression of vascular endothelial-cadherin (VE-cadherin) and occludin. In vitro, LHQK attenuated LPS-induced HPMECs injury by elevating TEER values and enhancing VE-cadherin and occludin protein levels. Finally, network pharmacology analysis and cellular level validation identified potential active ingredients of LHQK.ConclusionsIn summary, LHQK can mitigate LPS-induced inflammatory infiltration, pulmonary edema, and pulmonary vascular endothelial barrier dysfunction in the context of ALI/ARDS. This is achieved by decreasing the levels of VCAM-1, and increasing the expression levels of barrier-associated junctions, such as VE-cadherin and occludin. Consequently, LHQK exhibits promising therapeutic potential in preventing the progression of ALI/ARDS.

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Endothelial mitochondria determine rapid barrier failure in chemical lung injury.

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Endothelial mitochondria determine rapid barrier failure in chemical lung injury.

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