Fasudil alleviates LPS-induced lung injury by restoring aquaporin 5 expression and inhibiting inflammation in lungs
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摘要
Fasudil, a selective rho kinase(ROCK) inhibitor, has been reported to play a beneficial role in systemic inflammation in acute lung injury, but its mechanism for ameliorating pulmonary edema and inflammation remains unclear. Using hematoxylin-and-eosin(H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay,quantitative real time PCR and Western blotting, we found that fasudil attenuated LPS-induced lung injury, decreased lung edema, and suppressed inflammatory responses including leukocyte infiltration and IL-6 production. Further,fasudil upregulated LPS-induced aquaporin 5 reduction and inhibited NF-κB activation in the lungs of mice. Our results suggest that fasudil could restore the expression of aquaporin 5 to eliminate LPS-induced lung edema and prevent LPS-induced pulmonary inflammation by blocking the inflammatory pathway. Collectively, blockade of the ROCK pathway by fasudil may be a potential strategy for the treatment of acute lung injury.
Fasudil, a selective rho kinase(ROCK) inhibitor, has been reported to play a beneficial role in systemic inflammation in acute lung injury, but its mechanism for ameliorating pulmonary edema and inflammation remains unclear. Using hematoxylin-and-eosin(H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay,quantitative real time PCR and Western blotting, we found that fasudil attenuated LPS-induced lung injury, decreased lung edema, and suppressed inflammatory responses including leukocyte infiltration and IL-6 production. Further,fasudil upregulated LPS-induced aquaporin 5 reduction and inhibited NF-κB activation in the lungs of mice. Our results suggest that fasudil could restore the expression of aquaporin 5 to eliminate LPS-induced lung edema and prevent LPS-induced pulmonary inflammation by blocking the inflammatory pathway. Collectively, blockade of the ROCK pathway by fasudil may be a potential strategy for the treatment of acute lung injury.
Fasudil, a selective rho kinase(ROCK) inhibitor, has been reported to play a beneficial role in systemic inflammation in acute lung injury, but its mechanism for ameliorating pulmonary edema and inflammation remains unclear. Using hematoxylin-and-eosin(H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay,quantitative real time PCR and Western blotting, we found that fasudil attenuated LPS-induced lung injury, decreased lung edema, and suppressed inflammatory responses including leukocyte infiltration and IL-6 production. Further,fasudil upregulated LPS-induced aquaporin 5 reduction and inhibited NF-κB activation in the lungs of mice. Our results suggest that fasudil could restore the expression of aquaporin 5 to eliminate LPS-induced lung edema and prevent LPS-induced pulmonary inflammation by blocking the inflammatory pathway. Collectively, blockade of the ROCK pathway by fasudil may be a potential strategy for the treatment of acute lung injury.
引文
[1] Sapru A, Flori H, Quasney MW, et al., and the Pediatric Acute Lung Injury Consensus Conference Group. Pathobiology of acute respiratory distress syndrome. Pediatr Crit Care Med,2015, 16(5 Suppl 1):S6-S22.
[2] Standiford TJ, Ward PA. Therapeutic targeting of acute lung injury and acute respiratory distress syndrome. Transl Res,2016, 167(1):183-191.
[3] Matthay MA, Ware LB, Zimmerman GA. The acute respiratory distress syndrome. J Clin Invest, 2012, 122(8):2731-2740.
[4] Song Y, Jayaraman S, Yang B, et al. Role of aquaporin water channels in airway fluid transport, humidification, and surface liquid hydration. J Gen Physiol, 2001, 117(6):573-582.
[5] Dobbs LG, Gonzalez R, Matthay MA, et al. Highly waterpermeable type I alveolar epithelial cells confer high water permeability between the airspace and vasculature in rat lung.Proc Natl Acad Sci U S A, 1998, 95(6):2991-2996.
[6] Effros R M, Darin C, Jacobs E R, et al. Water transport and the distribution of aquaporin-1 in pulmonary air spaces. J Appl Physiol(1985), 1997, 83(3):1002-1016.
[7]Nielsen S, King LS, Christensen BM, et al. Aquaporins in complex tissues.Ⅱ. Subcellular distribution in respiratory and glandular tissues of rat Am J Physiol, 1997, 273(5 Pt 1):C1549-C1561.
[8] Song Y, Verkman AS. Aquaporin-5 dependent fluid secretion in airway submucosal glands. J Biol Chem, 2001, 276(44):41288-41292.
[9] Sun CY, Zhao YX, Zhong W, et al. The expression of aquaporins 1 and 5 in rat lung after thoracic irradiation. J Radiat Res, 2014, 55(4):683-689.
[10] Xu J, Huang B, Wang Y, et al. Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the upregulated expressions of AQP1 and AQP5 in lung. Clin Exp Pharmacol Physiol, 2016, 43(11):1071-1079.
[11] Towne JE, Harrod KS, Krane CM, et al. Decreased expression of aquaporin(AQP)1 and AQP5 in mouse lung after acute viralinfection. Am JRespir Cell Mol Biol, 2000, 22(1):34-44.
[12] Zhang ZQ, Song YL, Chen ZH, et al. Deletion of aquaporin 5aggravates acute lung injury induced by Pseudomonas aeruginosa. J Trauma, 2011, 71(5):1305-1311.
[13] Cernuda-Morollón E, Ridley AJ. Rho GTPases and leukocyte adhesion receptor expression and function in endothelial cells.Circ Res, 2006, 98(6):757-767.
[14] Li Y, Wu Y, Wang Z, et al. Fasudil attenuates lipopolysaccharide-induced acute lung injury in mice through the Rho/Rho kinase pathway. Med Sci Monit, 2010, 16(4):BR112-BR118.
[15] Ding RY, Zhao DM, Zhang ZD, et al. Pretreatment of Rho kinase inhibitor inhibits systemic inflammation and prevents endotoxin-induced acute lung injury in mice. J Surg Res, 2011,171(2):e209-e214.
[16] Sznajder JI. Alveolar edema must be cleared for the acute respiratory distress syndrome patient to survive. Am J Respir Crit Care Med, 2001, 163(6):1293-1294.
[17] Sartori C, Matthay MA. Alveolar epithelial fluid transport in acute lung injury:new insights. Eur Respir J, 2002, 20(5):1299-1313.
[18] Vadász I, Raviv S, Sznajder JI. Alveolar epithelium and Na,KATPase in acute lung injury. Intensive Care Med, 2007, 33(7):1243-1251.
[19] Verkman AS. Role of aquaporins in lung liquid physiology.Respir Physiol Neurobiol, 2007, 159(3):324-330.
[20] Walker D C, Mackenzie A L, Wiggs B R, et al. Assessment of tight junctions between pulmonary epithelial and endothelial cells. JAppl Physiol(1985), 1988, 64(6):2348-2356.
[21] McCarthy KM, Skare IB, Stankewich MC, et al. Occludin is a functional component of the tight junction.J Cell Sci, 1996,109(Pt 9):2287-2298.
[22] Chen Y, Merzdorf C, Paul DL, et al. COOH terminus of occludin is required for tight junction barrier function in early Xenopus embryos. J Cell Biol, 1997, 138(4):891-899.
[23] Guttman JA, Finlay BB. Tight junctions as targets of infectious agents. Biochim Biophys Acta, 2009, 1788(4):832-841.
[24] Capaldo CT, Nusrat A. Cytokine regulation of tight junctions.Biochim Biophys Acta, 2009, 1788(4):864-871.
[25] Schreibelt G, Kooij G, Reijerkerk A, et al. Reactive oxygen species alter brain endothelial tight junction dynamics via RhoA, PI3 kinase, and PKB signaling. FASEB J, 2007,21(13):3666-3676.
[26] Xu R, Feng X, Xie X, et al. HIV-1 Tat protein increases the permeability of brain endothelial cells by both inhibiting occludin expression and cleaving occludin via matrix metalloproteinase-9. Brain Res, 2012, 1436:13-19.
[27] Kawedia JD, Nieman ML, Boivin GP, et al. Interaction between transcellular and paracellular water transport pathways through Aquaporin 5 and the tight junction complex. Proc Natl Acad Sci U S A, 2007, 104(9):3621-3626.
[28] Dolinay T, Kim YS, Howrylak J, et al. Inflammasomeregulated cytokines are critical mediators of acute lung injury.Am J Respir Crit Care Med, 2012, 185(11):1225-1234.
[29] Mikacenic C, Hansen EE, Radella F, et al. Interleukin-17A Is Associated With Alveolar Inflammation and Poor Outcomes in Acute Respiratory Distress Syndrome. Crit Care Med, 2016,44(3):496-502.
[30] Mikkelsen M E, Shah C V, Scherpereel A, et al. Lower serum endocan levels are associated with the development of acute lung injury after major trauma. J Crit Care, 2012, 27(5):522e511-527.
[31] Strieter RM, Belperio JA, Keane MP. Cytokines in innate host defense in the lung. J Clin Invest, 2002, 109(6):699-705.
[32] Diamond G, Legarda D, Ryan LK. The innate immune response of the respiratory epithelium. Immunol Rev, 2000, 173:27-38.
[33] Mong PY, Wang Q. Activation of Rho kinase isoforms in lung endothelial cells during inflammation. J Immunol, 2009, 182(4):2385-2394.
[34] Li Q, Verma IM. NF-kappaB regulation in the immune system.Nat Rev Immunol, 2002, 2(10):725-734.
[35] Skerrett SJ, Liggitt HD, Hajjar AM, et al. Respiratory epithelial cells regulate lung inflammation in response to inhaled endotoxin. Am J Physiol Lung Cell Mol Physiol, 2004, 287(1):L143-L152.
[36] 17227815segain J P, Raingeard De La Bletiere D, Sauzeau V,et al. Rho kinase blockade prevents inflammation via nuclear factor kappa B inhibition:evidence in Crohn's disease and experimental colitis. Gastroenterology, 2003, 124(5):1180-1187.
[37] Yao C, Purwanti N, Karabasil MR, et al. Potential downregulation of salivary gland AQP5 by LPS via cross-coupling of NF-kappaB and p-c-Jun/c-Fos. Am J Pathol, 2010, 177(2):724-734.
[2] Standiford TJ, Ward PA. Therapeutic targeting of acute lung injury and acute respiratory distress syndrome. Transl Res,2016, 167(1):183-191.
[3] Matthay MA, Ware LB, Zimmerman GA. The acute respiratory distress syndrome. J Clin Invest, 2012, 122(8):2731-2740.
[4] Song Y, Jayaraman S, Yang B, et al. Role of aquaporin water channels in airway fluid transport, humidification, and surface liquid hydration. J Gen Physiol, 2001, 117(6):573-582.
[5] Dobbs LG, Gonzalez R, Matthay MA, et al. Highly waterpermeable type I alveolar epithelial cells confer high water permeability between the airspace and vasculature in rat lung.Proc Natl Acad Sci U S A, 1998, 95(6):2991-2996.
[6] Effros R M, Darin C, Jacobs E R, et al. Water transport and the distribution of aquaporin-1 in pulmonary air spaces. J Appl Physiol(1985), 1997, 83(3):1002-1016.
[7]Nielsen S, King LS, Christensen BM, et al. Aquaporins in complex tissues.Ⅱ. Subcellular distribution in respiratory and glandular tissues of rat Am J Physiol, 1997, 273(5 Pt 1):C1549-C1561.
[8] Song Y, Verkman AS. Aquaporin-5 dependent fluid secretion in airway submucosal glands. J Biol Chem, 2001, 276(44):41288-41292.
[9] Sun CY, Zhao YX, Zhong W, et al. The expression of aquaporins 1 and 5 in rat lung after thoracic irradiation. J Radiat Res, 2014, 55(4):683-689.
[10] Xu J, Huang B, Wang Y, et al. Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the upregulated expressions of AQP1 and AQP5 in lung. Clin Exp Pharmacol Physiol, 2016, 43(11):1071-1079.
[11] Towne JE, Harrod KS, Krane CM, et al. Decreased expression of aquaporin(AQP)1 and AQP5 in mouse lung after acute viralinfection. Am JRespir Cell Mol Biol, 2000, 22(1):34-44.
[12] Zhang ZQ, Song YL, Chen ZH, et al. Deletion of aquaporin 5aggravates acute lung injury induced by Pseudomonas aeruginosa. J Trauma, 2011, 71(5):1305-1311.
[13] Cernuda-Morollón E, Ridley AJ. Rho GTPases and leukocyte adhesion receptor expression and function in endothelial cells.Circ Res, 2006, 98(6):757-767.
[14] Li Y, Wu Y, Wang Z, et al. Fasudil attenuates lipopolysaccharide-induced acute lung injury in mice through the Rho/Rho kinase pathway. Med Sci Monit, 2010, 16(4):BR112-BR118.
[15] Ding RY, Zhao DM, Zhang ZD, et al. Pretreatment of Rho kinase inhibitor inhibits systemic inflammation and prevents endotoxin-induced acute lung injury in mice. J Surg Res, 2011,171(2):e209-e214.
[16] Sznajder JI. Alveolar edema must be cleared for the acute respiratory distress syndrome patient to survive. Am J Respir Crit Care Med, 2001, 163(6):1293-1294.
[17] Sartori C, Matthay MA. Alveolar epithelial fluid transport in acute lung injury:new insights. Eur Respir J, 2002, 20(5):1299-1313.
[18] Vadász I, Raviv S, Sznajder JI. Alveolar epithelium and Na,KATPase in acute lung injury. Intensive Care Med, 2007, 33(7):1243-1251.
[19] Verkman AS. Role of aquaporins in lung liquid physiology.Respir Physiol Neurobiol, 2007, 159(3):324-330.
[20] Walker D C, Mackenzie A L, Wiggs B R, et al. Assessment of tight junctions between pulmonary epithelial and endothelial cells. JAppl Physiol(1985), 1988, 64(6):2348-2356.
[21] McCarthy KM, Skare IB, Stankewich MC, et al. Occludin is a functional component of the tight junction.J Cell Sci, 1996,109(Pt 9):2287-2298.
[22] Chen Y, Merzdorf C, Paul DL, et al. COOH terminus of occludin is required for tight junction barrier function in early Xenopus embryos. J Cell Biol, 1997, 138(4):891-899.
[23] Guttman JA, Finlay BB. Tight junctions as targets of infectious agents. Biochim Biophys Acta, 2009, 1788(4):832-841.
[24] Capaldo CT, Nusrat A. Cytokine regulation of tight junctions.Biochim Biophys Acta, 2009, 1788(4):864-871.
[25] Schreibelt G, Kooij G, Reijerkerk A, et al. Reactive oxygen species alter brain endothelial tight junction dynamics via RhoA, PI3 kinase, and PKB signaling. FASEB J, 2007,21(13):3666-3676.
[26] Xu R, Feng X, Xie X, et al. HIV-1 Tat protein increases the permeability of brain endothelial cells by both inhibiting occludin expression and cleaving occludin via matrix metalloproteinase-9. Brain Res, 2012, 1436:13-19.
[27] Kawedia JD, Nieman ML, Boivin GP, et al. Interaction between transcellular and paracellular water transport pathways through Aquaporin 5 and the tight junction complex. Proc Natl Acad Sci U S A, 2007, 104(9):3621-3626.
[28] Dolinay T, Kim YS, Howrylak J, et al. Inflammasomeregulated cytokines are critical mediators of acute lung injury.Am J Respir Crit Care Med, 2012, 185(11):1225-1234.
[29] Mikacenic C, Hansen EE, Radella F, et al. Interleukin-17A Is Associated With Alveolar Inflammation and Poor Outcomes in Acute Respiratory Distress Syndrome. Crit Care Med, 2016,44(3):496-502.
[30] Mikkelsen M E, Shah C V, Scherpereel A, et al. Lower serum endocan levels are associated with the development of acute lung injury after major trauma. J Crit Care, 2012, 27(5):522e511-527.
[31] Strieter RM, Belperio JA, Keane MP. Cytokines in innate host defense in the lung. J Clin Invest, 2002, 109(6):699-705.
[32] Diamond G, Legarda D, Ryan LK. The innate immune response of the respiratory epithelium. Immunol Rev, 2000, 173:27-38.
[33] Mong PY, Wang Q. Activation of Rho kinase isoforms in lung endothelial cells during inflammation. J Immunol, 2009, 182(4):2385-2394.
[34] Li Q, Verma IM. NF-kappaB regulation in the immune system.Nat Rev Immunol, 2002, 2(10):725-734.
[35] Skerrett SJ, Liggitt HD, Hajjar AM, et al. Respiratory epithelial cells regulate lung inflammation in response to inhaled endotoxin. Am J Physiol Lung Cell Mol Physiol, 2004, 287(1):L143-L152.
[36] 17227815segain J P, Raingeard De La Bletiere D, Sauzeau V,et al. Rho kinase blockade prevents inflammation via nuclear factor kappa B inhibition:evidence in Crohn's disease and experimental colitis. Gastroenterology, 2003, 124(5):1180-1187.
[37] Yao C, Purwanti N, Karabasil MR, et al. Potential downregulation of salivary gland AQP5 by LPS via cross-coupling of NF-kappaB and p-c-Jun/c-Fos. Am J Pathol, 2010, 177(2):724-734.
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