IL-10 inhibits inflammatory cytokines released by fetal mouse lung fibroblasts exposed to mechanical stretch
Renda L. Hawwa
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorMichael A. Hokenson
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorYulian Wang
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorZheping Huang
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorSurendra Sharma
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorCorresponding Author
Juan Sanchez-Esteban MD
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, RI 02905.Search for more papers by this authorRenda L. Hawwa
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorMichael A. Hokenson
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorYulian Wang
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorZheping Huang
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorSurendra Sharma
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Search for more papers by this authorCorresponding Author
Juan Sanchez-Esteban MD
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, Rhode Island 02905
Warren Alpert Medical School of Brown University, Providence, Rhode Island 02905
Department of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, RI 02905.Search for more papers by this authorAbstract
Background
Mechanical ventilation plays an important role in the pathogenesis of bronchopulmonary dysplasia. However, the molecular mechanisms by which excessive stretch induces lung inflammation are not well characterized.
Objectives
In this study, we investigated in vitro the contribution of lung mesenchymal cells to the inflammatory response mediated by mechanical stretch and the potential protective role of IL-10.
Methods
Fetal mouse lung fibroblasts isolated during the saccular stage of lung development were exposed to 20% cyclic stretch to simulate mechanical injury. The phenotype of cultured fibroblasts was investigated by red oil O and alpha-smooth muscle actin (α-SMA) staining. Cell necrosis, apoptosis, and inflammation were analyzed by lactate dehydrogenase release, cleaved caspase-3 activation and release of cytokines and chemokines into the supernatant, respectively.
Results
First, we characterized the phenotype of the cultured fibroblasts and found an absence of red oil O staining and 100% positive staining for α-SMA, indicating that cultured fibroblasts were myofibroblasts. Mechanical stretch increased necrosis and apoptosis by two- and three-fold, compared to unstretched samples. Incubation of monolayers with IL-10 prior to stretch did not affect necrosis but significantly decreased apoptosis. Mechanical stretch increased release of pro-inflammatory cytokines and chemokines IL-1β, MCP-1, RANTES, IL-6, KC and TNF-α into the supernatant by 1.5- to 2.5-fold, and administration of IL-10 before stretch blocked that release.
Conclusions
Our data demonstrate that lung interstitial cells may play a significant role in the inflammatory cascade triggered by mechanical stretch. IL-10 protects fetal fibroblasts from injury secondary to stretch. Pediatr. Pulmonol. 2011; 46:640–649. © 2011 Wiley-Liss, Inc.
REFERENCES
- 1 Tibboel D, Jobe AH. Update in pediatric lung disease 2009. Am J Respir Crit Care Med 2010; 181: 661–665.
- 2 Carlton DP, Albertine KH, Cho SC, Lont M, Bland RD. Role of neutrophils in lung vascular injury and edema after premature birth in lambs. J Appl Physiol 1997; 83: 1307–1317.
- 3 Speer CP. Inflammation and bronchopulmonary dysplasia: A continuing story. Semin Fetal Neonat Med 2006; 11: 354–362.
- 4 Speer CP. Inflammation and bronchopulmonary dysplasia. Semin Neonatal 2003; 8: 29–38.
- 5 Hammerschmidt S, Kuhn H, Sack U, Schlenska A, Gessner C, Gillissen A, Wirtz H. Mechanical stretch alters alveolar type II cell mediator release toward a proinflammatory pattern. Am J Respir Cell Mol Biol 2005; 33: 203–210.
- 6 Lee HS, Wang Y, Maciejewski BS, Esho K, Fulton C, Sharma S, Sanchez-Esteban J. Interleukin-10 protects cultured fetal rat type II epithelial cells from injury induced by mechanical stretch. Am J Physiol Lung Cell Mol Physiol 2008; 294: L225–L232.
- 7 Vlahakis NE, Schroeder MA, Limper AH, Hubmayr RD. Stretch induces cytokine release by alveolar epithelial cells in vitro. Am J Physiol 1999; 277: L167–L173.
- 8 Thorley AJ, Ford PA, Giembycz MA, Goldstraw P, Young A, Tetley TD. Differential regulation of cytokine release and leukocyte migration by lipopolysaccharide-stimulated primary human lung alveolar type II epithelial cells and macrophages. J Immunol 2007; 178: 463–473.
- 9 McGowan SE, Torday JS. The pulmonary lipofibroblast (lipid interstitial cell) and its contributions to alveolar development. Annu Rev Physiol 1997; 59: 43–62.
- 10 Gahler A, Stallmach T, Schwaller J, Fey MF, Tobler A. Interleukin-8 expression by fetal and neonatal pulmonary cells in hyaline membrane disease and amniotic infection. Pediatr Res 2000; 48: 299–303.
- 11 Opal SM, DePalo VA. Anti-inflammatory cytokines. Chest 2000; 117: 1162–1172.
- 12 Thaxton JE, Sharma S. Interleukin-10: A multi-faceted agent of pregnancy. Am J Reprod Immunol 2010; 63: 482–491.
- 13 Kwong KY, Jones CA, Cayabyab R, Lecart C, Khuu N, Rhandhawa I, Hanley JM, Ramanathan R, deLemos RA, Minoo P. The effects of IL-10 on proinflammatory cytokine expression (IL-1beta and IL-8) in hyaline membrane disease (HMD). Clin Immunol Immunopathol 1998; 88: 105–113.
- 14 Oei J, Lui K, Wang H, Henry R. Decreased interleukin-10 in tracheal aspirates from preterm infants developing chronic lung disease. Acta Paediatr 2002; 91: 1194–1199.
- 15 Schultz C, Tautz J, Reiss I, Moller JC. Prolonged mechanical ventilation induces pulmonary inflammation in preterm infants. Biol Neonate 2003; 84: 64–66.
- 16 McGowan EC, Kostadinov S, McLean K, Gotsch F, Venturini D, Romero R, Laptook AR, Sharma S. Placental IL-10 dysregulation and association with bronchopulmonary dysplasia risk. Pediatr Res 2009; 66: 455–460.
- 17 Blahnik MJ, Ramanathan R, Riley CR, Minoo P. Lipopolysaccharide-induced tumor necrosis factor-alpha and IL-10 production by lung macrophages from preterm and term neonates. Pediatr Res 2001; 50: 726–731.
- 18 Garingo A, Tesoriero L, Cayabyab R, Durand M, Blahnik M, Sardesai S, Ramanathan R, Jones C, Kwong K, Li C, et al. Constitutive IL-10 expression by lung inflammatory cells and risk for bronchopulmonary dysplasia. Pediatr Res 2007; 61: 197–202.
- 19 Jones CA, Cayabyab RG, Kwong KY, Stotts C, Wong B, Hamdan H, Minoo P. deLemos RA. Undetectable interleukin (IL)-10 and persistent IL-8 expression early in hyaline membrane disease: A possible developmental basis for the predisposition to chronic lung inflammation in preterm newborns. Pediatr Res 1996; 39: 966–975.
- 20 Marchant A, Bruyns C, Vandenabeele P, Ducarme M, Gerard C, Delvaux A, De Groote D, Abramowicz D, Velu T, Goldman M. Interleukin-10 controls interferon-gamma and tumor necrosis factor production during experimental endotoxemia. Eur J Immunol 1994; 24: 1167–1171.
- 21 Wang Y, Maciejewski BS, Soto-Reyes D, Lee HS, Warburton D, Sanchez-Esteban J. Mechanical stretch promotes fetal type II epithelial cell differentiation via shedding of HB-EGF and TGF-alpha. J Physiol 2009; 587: 1739–1753.
- 22 Tschumperlin DJ, Oswari J, Margulies AS. Deformation-induced injury of alveolar epithelial cells. Effect of frequency, duration, and amplitude. Am J Respir Crit Care Med 2000; 162: 357–362.
- 23 Wang Y, Maciejewski BS, Drouillard D, Santos M, Hokenson MA, Hawwa RL, Huang Z, Sanchez-Esteban J. A role for caveolin-1 in mechanotransduction of fetal type II epithelial cells. Am J Physiol Lung Cell Mol Physiol 2010; 298: L775–L783.
- 24 Wang Y, Maciejewski BS, Lee N, Silbert O, McKnight NL, Frangos JA, Sanchez-Esteban J. Strain-induced fetal type II epithelial cell differentiation is mediated via cAMP-PKA-dependent signaling pathway. Am J Physiol Lung Cell Mol Physiol 2006; 291: L820–827.
- 25 Rehan V, Torday J. Hyperoxia augments pulmonary lipofibroblast-to-myofibroblast transdifferentiation. Cell Biochem Biophys 2003; 38: 239–250.
- 26 Bland RD, Ertsey R, Mokres LM, Xu L, Jacobson BE, Jiang S, Alvira CM, Rabinovitch M, Shinwell ES, Dixit A. Mechanical ventilation uncouples synthesis and assembly of elastin and increases apoptosis in lungs of newborn mice. Prelude to defective alveolar septation during lung development? Am J Physiol Lung Cell Mol Physiol 2008; 294: L3–L14.
- 27 Bland RD, Mokres LM, Ertsey R, Jacobson BE, Jiang S, Rabinovitch M, Xu L, Shinwell ES, Zhang F, Beasley MA. Mechanical ventilation with 40% oxygen reduces pulmonary expression of genes that regulate lung development and impairs alveolar septation in newborn mice. Am J Physiol Lung Cell Mol Physiol 2007; 293: L1099–L1110.
- 28 Mokres LM, Parai K, Hilgendorff A, Ertsey R, Alvira CM, Rabinovitch M, Bland RD. Prolonged mechanical ventilation with air induces apoptosis and causes failure of alveolar septation and angiogenesis in lungs of newborn mice. Am J Physiol Lung Cell Mol Physiol 2010; 298: L23–L35.
- 29 Darby IA, Hewitson TD. Fibroblast differentiation in wound healing and fibrosis. Int Rev Cytol 2007; 257: 143–179.
- 30 Bruce MC, Honaker CE, Cross RJ. Lung fibroblasts undergo apoptosis following alveolarization. Am J Respir Cell Mol Biol 1999; 20: 228–236.
- 31 Gessner AM. Transdifferentiation of hepatic stellate cells (Ito cells) to myofibroblasts: A key event in hepatic fibrogenesis. Kidney Int Suppl 1996; 54: S39–S45.
- 32 Vlahakis NE, Hubmayr RD. Cellular stress failure in ventilator-injured lungs. Am J Respir Crit Care Med 2005; 171: 1328–1342.
- 33 Hammerschmidt S, Kuhn H, Grasenack T, Gessner C, Wirtz H. Apoptosis and necrosis induced by cyclic mechanical stretching in alveolar type II cells. Am J Respir Cell Mol Biol 2004; 30: 396–402.
- 34 Mourgeon E, Isowa N, Keshavjee S, Zhang X, Slutsky AS, Liu M. Mechanical stretch stimulates macrophage inflammatory protein-2 secretion from fetal rat lung cells. Am J Physiol Lung Cell Mol Physiol 2000; 279: L699–L706.
- 35 Edwards YS, Sutherland LM, Power JH, Nicholas TE, Murray AW. Cyclic stretch induces both apoptosis and secretion in rat alveolar type II cells. FEBS Lett 1999; 448: 127–130.
- 36 De Paepe ME, Sardesai MP, Johnson BD, Lesieur-Brooks AM, Papadakis K, Luks FI. The role of apoptosis in normal and accelerated lung development in fetal rabbits. J Pediatr Surg 1999; 34: 863–870, discussion 870–861.
- 37 Bustani P, Hodge R, Tellabati A, Li J, Pandya H, Kotecha S. Differential response of the epithelium and interstitium in developing human fetal lung explants to hyperoxia. Pediatr Res 2006; 59: 383–388.
- 38 Sanchez-Esteban J, Wang Y, Cicchiello LA, Rubin LP. Cyclic mechanical stretch inhibits cell proliferation and induces apoptosis in fetal rat lung fibroblasts. Am J Physiol Lung Cell Mol Physiol 2002; 282: L448–L456.
- 39 Numanami H, Koyama S, Sato E, Haniuda M, Nelson DK, Hoyt JC, Freels JL, Habib MP, Robbins RA. Serine protease inhibitors modulate chemotactic cytokine production by human lung fibroblasts in vitro. Am J Physiol Lung Cell Mol Physiol 2003; 284: L882–L890.
- 40 Stancombe BB, Walsh WF, Derdak S, Dixon P, Hensley D. Induction of human neonatal pulmonary fibroblast cytokines by hyperoxia and Ureaplasma urealyticum. Clin Infect Dis 1993; S154–S157.
- 41 Le Bellego F, Perera H, Plante S, Chakir J, Hamid Q, Ludwig MS. Mechanical strain increases cytokine and chemokine production in bronchial fibroblasts from asthmatic patients. Allergy 2009; 64: 32–39.
- 42
Haitsma JJ,
Schultz MJ,
Slutsky AS.
Cytokin release. In:
PJ Papadakos,
B Lachmann, editors.
Mechanical ventilation: Clinical applications and pathophysiology.
Philadelphia, PA:
Saunders, Elsevier, Inc;
2008. pp.
216–224.
10.1016/B978-0-7216-0186-1.50023-5 Google Scholar
- 43 Davidson D, Miskolci V, Clark DC, Dolmaian G, Vancurova I. Interleukin-10 production after pro-inflammatory stimulation of neutrophils and monocytic cells of the newborn. Comparison to exogenous interleukin-10 and dexamethasone levels needed to inhibit chemokine release. Neonatology 2007; 92: 127–133.
- 44 Tryzmel J, Miskolci V, Castro-Alcaraz S, Vancurova I, Davidson D. Interleukin-10 inhibits proinflammatory chemokine release by neutrophils of the newborn without suppression of nuclear factor-kappa B. Pediatr Res 2003; 54: 382–386.
- 45 Mulligan MS, Warner RL, Foreback JL, Shanley TP, Ward PA. Protective effects of IL-4, IL-10, IL-12, and IL-13 in IgG immune complex-induced lung injury: Role of endogenous IL-12. J Immunol 1997; 159: 3483–3489.
- 46 McAuliffe PF, Murday ME, Efron PA, Scumpia PO, Ungaro R, Abouhamze A, Tannahill CL, Hutchins B, LaFace D, Moldawer LL. Dose-dependent improvements in outcome with adenoviral expression of interleukin-10 in a murine model of multisystem organ failure. Gene Therapy 2006; 13: 276–282.
- 47 Hoegl S, Boost KA, Czerwonka H, Dolfen A, Scheiermann P, Muhl H, Zwissler B, Hofstetter C. Inhaled IL-10 reduces biotrauma and mortality in a model of ventilator-induced lung injury. Respir Med 2009; 103: 463–470.
Citing Literature
