miR-144–mediated Inhibition of ROCK1 Protects against LPS-induced Lung Endothelial Hyperpermeability
| Autor: | Suhail Akhtar, Mohd Shahid, Mohammad Tauseef, Thomas P. Shanley, M. Rizwan Siddiqui, Kelli McDonough |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Lipopolysaccharides
Male 0301 basic medicine Pulmonary and Respiratory Medicine Clinical Biochemistry Inflammation Vascular permeability Lung injury Permeability Proinflammatory cytokine Mice Myosin-Light-Chain Phosphatase 03 medical and health sciences 0302 clinical medicine Electric Impedance medicine Animals Humans Lung cancer Lung Molecular Biology rho-Associated Kinases Chemistry Microcirculation Endothelial Cells Cell Biology medicine.disease Pulmonary edema Mice Inbred C57BL Endothelial stem cell MicroRNAs 030104 developmental biology medicine.anatomical_structure 030228 respiratory system Reperfusion Injury Liposomes Cancer research medicine.symptom Signal Transduction |
| Zdroj: | American Journal of Respiratory Cell and Molecular Biology. 61:257-265 |
| ISSN: | 1535-4989 1044-1549 |
| DOI: | 10.1165/rcmb.2018-0235oc |
| Popis: | Dysfunctional endothelial cell (EC) barrier and increased lung vascular permeability is a cardinal feature of acute lung injury and sepsis that may result in a pathophysiological condition characterized by alveolar flooding, pulmonary edema, and subsequent hypoxemia. In lung ECs, activation of Rho-associated kinase-1 (ROCK1) phosphorylates myosin light chain (MLC)-associated phosphatase at its inhibitory site, which favors phosphorylation of MLC, stress fiber formation, and hyperpermeability during acute lung injury. The role of microRNA-144 (miR-144) has been well investigated in many human diseases, including cardiac ischemia/reperfusion-induced injury, lung cancer, and lung viral infection; however, its role in pulmonary EC barrier regulation remains obscure. Here, we investigated the miR-144-mediated mechanism in the protection of endothelial barrier function in an LPS-induced lung injury model. By using transendothelial electrical resistance and transwell permeability assay to examine in vitro permeability and immunofluorescence microscopy to determine barrier integrity, we showed that ectopic expression of miR-144 effectively blocked lung EC barrier disruption and hyperpermeability in response to proinflammatory agents. Furthermore, using a gain-and-loss-of-function strategy, overexpression of miR-144 significantly decreased ROCK1 expression. Concomitantly, miR-144 inhibits ROCK1-mediated phosphorylation of MLC phosphataseThr853 and thus phosphorylation of MLCThr18/Ser19 to counteract stress fiber formation in LPS-activated EC. Finally, in LPS-challenged mice, intranasal delivery of miR-144 mimic via liposomes attenuated endotoxemia-induced increases in lung wet/dry ratio, vascular permeability, and inflammation. In conclusion, these data suggest that miR-144-attenuated activation of inflammatory ROCK1/MLC pathway in vascular ECs is a promising therapeutic strategy to counter inflammatory lung injury. |
| Databáze: | OpenAIRE |
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