Hyperoxia disrupts pulmonary epithelial barrier in newborn rats via the deterioration of occludin and ZO-1

Respiratory Research

Xindong Xue²  Zhiling Yu²  Xiaohong Yue¹  Shuyan Xu²  Jianhua Fu²  Xuewen Xu³  Kai You² 

[1] Department of Pediatrics, Fengtian Hospital Affiliated to Shenyang Medical College, Shenyang, 110024, China;Department of Pediatrics, Shengjing Hospital of China Medical University, Shenyang, 110004, China;Department of Urology, Shengjing Hospital of China Medical University, Shenyang, 110004, China

关键词: Tight Junction;    Permeability;    Newborn;    Hyperoxia;    Acute lung injury;   

Others  :  796726
DOI  :  10.1186/1465-9921-13-36

 received in 2012-02-03, accepted in 2012-05-04,  发布年份 2012

【 摘 要 】

Background

Prolonged exposure to hyperoxia in neonates can cause hyperoxic acute lung injury (HALI), which is characterized by increased pulmonary permeability and diffuse infiltration of various inflammatory cells. Disruption of the epithelial barrier may lead to altered pulmonary permeability and maintenance of barrier properties requires intact epithelial tight junctions (TJs). However, in neonatal animals, relatively little is known about how the TJ proteins are expressed in the pulmonary epithelium, including whether expression of TJ proteins is regulated in response to hyperoxia exposure. This study determines whether changes in tight junctions play an important role in disruption of the pulmonary epithelial barrier during hyperoxic acute lung injury.

Methods

Newborn rats, randomly divided into two groups, were exposed to hyperoxia (95% oxygen) or normoxia for 1–7 days, and the severity of lung injury was assessed; location and expression of key tight junction protein occludin and ZO-1 were examined by immunofluorescence staining and immunobloting; messenger RNA in lung tissue was studied by RT-PCR; transmission electron microscopy study was performed for the detection of tight junction morphology.

Results

We found that different durations of hyperoxia exposure caused different degrees of lung injury in newborn rats. Treatment with hyperoxia for prolonged duration contributed to more serious lung injury, which was characterized by increased wet-to-dry ratio, extravascular lung water content, and bronchoalveolar lavage fluid (BALF):serum FD4 ratio. Transmission electron microscopy study demonstrated that hyperoxia destroyed the structure of tight junctions and prolonged hyperoxia exposure, enhancing the structure destruction. The results were compatible with pathohistologic findings. We found that hyperoxia markedly disrupted the membrane localization and downregulated the cytoplasm expression of the key tight junction proteins occludin and ZO-1 in the alveolar epithelium by immunofluorescence. The changes of messenger RNA and protein expression of occludin and ZO-1 in lung tissue detected by RT-PCR and immunoblotting were consistent with the degree of lung injury.

Conclusions

These data suggest that the disruption of the pulmonary epithelial barrier induced by hyperoxia is, at least in part, due to massive deterioration in the expression and localization of key TJ proteins.

【 授权许可】

   
2012 You et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Jobe AH, Bancalari E: Bronchopulmonary dysplasia. Am J Respir Crit Care Med 2001, 163:1723-1729.
• [2]Bancalari E, Claure N, Sosenko IR: Bronchopulmonary dysplasia: changes in pathogenesis, epidemiology and definition. Semin Neonatol 2003, 8:63-71.
• [3]Smith VC, Zupancic JA, McCormick MC, Croen LA, Greene J, Escobar GJ, Richardson DK: Rehospitalization in the first year of life among infants with bronchopulmonary dysplasia. J Pediatr 2004, 144:799-803.
• [4]Doyle LW: Respiratory function at age 8–9 years in extremely low birthweight/very preterm children born in Victoria in 1991–1992. Pediatr Pulmonol 2006, 41:570-576.
• [5]Doyle LW, Faber B, Callanan C, Freezer N, Ford GW, Davis NM: Bronchopulmonary dysplasia in very low birth weight subjects and lung function in late adolescence. Pediatrics 2006, 118:108-113.
• [6]Short EJ, Klein NK, Lewis BA, Fulton S, Eisengart S, Kercsmar C, Baley J, Singer LT: Cognitive and academic consequences of bronchopulmonary dysplasia and very low birth weight: 8-year-old outcomes. Pediatrics 2003, 112:e359.
• [7]Anderson PJ, Doyle LW: Neurodevelopmental outcome of bronchopulmonary dysplasia. Semin Perinatol 2006, 30:227-232.
• [8]Kotecha S, Chan B, Azam N, Silverman M, Shaw RJ: Increase in interleukin-8 and soluble intercellular adhesion molecule-1 in bronchoalveolar lavage fluid from premature infants who develop chronic lung disease. Arch Dis Child Fetal Neonatal Ed 1995, 72:F90-F96.
• [9]Coalson JJ: Pathology of new bronchopulmonary dysplasia. Semin Neonatol 2003, 8:73-81.
• [10]Warner BB, Stuart LA, Papes RA, Wispe JR: Functional and pathological effects of prolonged hyperoxia in neonatal mice. Am J Physiol 1998, 275:L110-L117.
• [11]Corne J, Chupp G, Lee CG, Homer RJ, Zhu Z, Chen Q, Ma B, Du Y, Roux F, McArdle J, et al.: IL-13 stimulates vascular endothelial cell growth factor and protects against hyperoxic acute lung injury. J Clin Invest 2000, 106:783-791.
• [12]Altemeier WA, Sinclair SE: Hyperoxia in the intensive care unit: why more is not always better. Curr Opin Crit Care 2007, 13:73-78.
• [13]Crandall ED, Matthay MA: Alveolar epithelial transport. Basic science to clinical medicine. Am J Respir Crit Care Med 2001, 163:1021-1029.
• [14]Ware LB, Matthay MA: Alveolar fluid clearance is impaired in the majority of patients with acute lung injury and the acute respiratory distress syndrome. Am J Respir Crit Care Med 2001, 163:1376-1383.
• [15]Schneeberger EE, Lynch RD: Structure, function, and regulation of cellular tight junctions. Am J Physiol 1992, 262:L647-L661.
• [16]Gonzalez-Mariscal L, Betanzos A, Nava P, Jaramillo BE: Tight junction proteins. Prog Biophys Mol Biol 2003, 81:1-44.
• [17]McCarthy KM, Skare IB, Stankewich MC, Furuse M, Tsukita S, Rogers RA, Lynch RD, Schneeberger EE: Occludin is a functional component of the tight junction. J Cell Sci 1996, 109(Pt 9):2287-2298.
• [18]Chen Y, Merzdorf C, Paul DL, Goodenough DA: COOH terminus of occludin is required for tight junction barrier function in early Xenopus embryos. J Cell Biol 1997, 138:891-899.
• [19]Denker BM, Nigam SK: Molecular structure and assembly of the tight junction. Am J Physiol 1998, 274:F1-F9.
• [20]Ohtake K, Maeno T, Ueda H, Ogihara M, Natsume H, Morimoto Y: Poly-L-arginine enhances paracellular permeability via serine/threonine phosphorylation of ZO-1 and tyrosine dephosphorylation of occludin in rabbit nasal epithelium. Pharm Res 2003, 20:1838-1845.
• [21]Ye L, Martin TA, Parr C, Harrison GM, Mansel RE, Jiang WG: Biphasic effects of 17-beta-estradiol on expression of occludin and transendothelial resistance and paracellular permeability in human vascular endothelial cells. J Cell Physiol 2003, 196:362-369.
• [22]Chasiotis H, Wood CM, Kelly SP: Cortisol reduces paracellular permeability and increases occludin abundance in cultured trout gill epithelia. Mol Cell Endocrinol 2010, 323:232-238.
• [23]Raleigh DR, Boe DM, Yu D, Weber CR, Marchiando AM, Bradford EM, Wang Y, Wu L, Schneeberger EE, Shen L, Turner JR: Occludin S408 phosphorylation regulates tight junction protein interactions and barrier function. J Cell Biol 2011, 193:565-582.
• [24]Han X, Fink MP, Yang R, Delude RL: Increased iNOS activity is essential for intestinal epithelial tight junction dysfunction in endotoxemic mice. Shock 2004, 21:261-270.
• [25]Noth R, Lange-Grumfeld J, Stuber E, Kruse ML, Ellrichmann M, Hasler R, Hampe J, Bewig B, Rosenstiel P, Schreiber S, Arlt A: Increased intestinal permeability and tight junction disruption by altered expression and localization of occludin in a murine graft versus host disease model. BMC Gastroenterol 2011, 11:109. BioMed Central Full Text
• [26]Gon Y, Wood MR, Kiosses WB, Jo E, Sanna MG, Chun J, Rosen H: S1P3 receptor-induced reorganization of epithelial tight junctions compromises lung barrier integrity and is potentiated by TNF. Proc Natl Acad Sci U S A 2005, 102:9270-9275.
• [27]Mazzon E, Cuzzocrea S: Role of TNF-alpha in lung tight junction alteration in mouse model of acute lung inflammation. Respir Res 2007, 8:75. BioMed Central Full Text
• [28]Zhang YL, Li QQ, Guo W, Huang Y, Yang J: Effects of chronic ethanol ingestion on tight junction proteins and barrier function of alveolar epithelium in the rat. Shock 2007, 28:245-252.
• [29]Matute-Bello G, Downey G, Moore BB, Groshong SD, Matthay MA, Slutsky AS, Kuebler WM: An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals. Am J Respir Cell Mol Biol 2011, 44:725-738.
• [30]Li T, Koshy S, Folkesson HG: Involvement of {alpha}ENaC and Nedd4-2 in the conversion from lung fluid secretion to fluid absorption at birth in the rat as assayed by RNA interference analysis. Am J Physiol Lung Cell Mol Physiol 2007, 293:L1069-L1078.
• [31]Song Y, Fukuda N, Bai C, Ma T, Matthay MA, Verkman AS: Role of aquaporins in alveolar fluid clearance in neonatal and adult lung, and in oedema formation following acute lung injury: studies in transgenic aquaporin null mice. J Physiol 2000, 525(Pt 3):771-779.
• [32]Ben-Ari J, Makhoul IR, Dorio RJ, Buckley S, Warburton D, Walker SM: Cytokine response during hyperoxia: sequential production of pulmonary tumor necrosis factor and interleukin-6 in neonatal rats. Isr Med Assoc J 2000, 2:365-369.
• [33]Auten RL, Davis JM: Oxygen toxicity and reactive oxygen species: the devil is in the details. Pediatr Res 2009, 66:121-127.
• [34]Doyle LW, Ehrenkranz RA, Halliday HL: Dexamethasone treatment in the first week of life for preventing bronchopulmonary dysplasia in preterm infants: a systematic review. Neonatology 2010, 98:217-224.
• [35]Dani C, Cecchi A, Bertini G: Role of oxidative stress as physiopathologic factor in the preterm infant. Minerva Pediatr 2004, 56:381-394.
• [36]Comhair SA, Erzurum SC: Antioxidant responses to oxidant-mediated lung diseases. Am J Physiol Lung Cell Mol Physiol 2002, 283:L246-L255.
• [37]Deng H, Mason SN, Auten RL: Lung inflammation in hyperoxia can be prevented by antichemokine treatment in newborn rats. Am J Respir Crit Care Med 2000, 162:2316-2323.
• [38]Reutershan J, Basit A, Galkina EV, Ley K: Sequential recruitment of neutrophils into lung and bronchoalveolar lavage fluid in LPS-induced acute lung injury. Am J Physiol Lung Cell Mol Physiol 2005, 289:L807-L815.
• [39]Jankov RP, Johnstone L, Luo X, Robinson BH, Tanswell AK: Macrophages as a major source of oxygen radicals in the hyperoxic newborn rat lung. Free Radic Biol Med 2003, 35:200-209.
• [40]Yi M, Jankov RP, Belcastro R, Humes D, Copland I, Shek S, Sweezey NB, Post M, Albertine KH, Auten RL, Tanswell AK: Opposing effects of 60% oxygen and neutrophil influx on alveologenesis in the neonatal rat. Am J Respir Crit Care Med 2004, 170:1188-1196.
• [41]Ahamed K, Epaud R, Holzenberger M, Bonora M, Flejou JF, Puard J, Clement A, Henrion-Caude A: Deficiency in type 1 insulin-like growth factor receptor in mice protects against oxygen-induced lung injury. Respir Res 2005, 6:31. BioMed Central Full Text
• [42]Lozon TI, Eastman AJ, Matute-Bello G, Chen P, Hallstrand TS, Altemeier WA: PKR-dependent CHOP induction limits hyperoxia-induced lung injury. Am J Physiol Lung Cell Mol Physiol 2011, 300:L422-L429.
• [43]Wang M, Luo Z, Liu S, Li L, Deng X, Huang F, Shang L, Jian C, Yue S: Glutamate mediates hyperoxia-induced newborn rat lung injury through N-methyl-D-aspartate receptors. Am J Respir Cell Mol Biol 2009, 40:260-267.
• [44]Aggarwal NR, D'Alessio FR, Tsushima K, Files DC, Damarla M, Sidhaye VK, Fraig MM, Polotsky VY, King LS: Moderate oxygen augments lipopolysaccharide-induced lung injury in mice. Am J Physiol Lung Cell Mol Physiol 2010, 298:L371-L381.
• [45]Rao RK, Basuroy S, Rao VU, Karnaky KJ, Gupta A: Tyrosine phosphorylation and dissociation of occludin-ZO-1 and E-cadherin-beta-catenin complexes from the cytoskeleton by oxidative stress. Biochem J 2002, 368:471-481.