Background Nitric oxide (NO) plays an important role in mobilization of endothelial progenitor cells (EPCs). We hypothesized that inhaled NO (iNO) would induce EPC mobilization and therefore promote lung repair in acute respiratory distress syndrome (ARDS). Methodology/Principal Findings Healthy piglets were randomized into four groups (n = 6): Control (Con; mechanical ventilation only); ARDS (established by oleic acid infusion and mechanical ventilation); ARDS plus granulocyte-colony stimulating factor (G-CSF; 10 μg/kg/d subcutaneously); ARDS plus NO inhalation (iNO; 10 ppm). EPCs and mobilizing cytokines were assayed at different time points (baseline, 0, 24, 72 and 168 h) and injury reparation was assessed at 168 h. Compared to the Con group, the levels of EPCs were increased in bone marrow but not in blood in the ARDS group at 24 h. Compared to the ARDS group, inhaled NO induced a rapid elevation in the number of CD34+KDR+, KDR+CD133+ and CD34+KDR+CD133+ EPCs in blood (2163±454 vs. 1094±416, 1302±413 vs. 429±244, 1140±494 vs. 453±273 cells/ml, respectively, P<0.05), and a reduction in the percentage of KDR+CD133+ cells in bone marrow. Lung CD34, CD133, VEGF, VEGF receptor 2, endothelial NO synthase mRNA, and VEGF and VEGF receptor 2 protein expression levels were augmented in the iNO group, but not in the G-CSF group, compared to ARDS. Furthermore, iNO treatment reduced vascular permeability, increased pulmonary vessel density, and alleviated pulmonary edema and inflammation compared to ARDS treatment. Plasma VEGF, stromal cell-derived factor-1 (SDF-1) and bone marrow NO2？/NO3？ were significantly higher in the iNO group compared to the ARDS group at 72 h. Conclusions These results suggest that iNO induces mobilization of EPCs from bone marrow into circulation, contributes to vascular repair, and thereby alleviates lung damage.
References
[1]
Ware LB, Matthay MA (2000) The acute respiratory distress syndrome. N Engl J Med 342: 1334–1349.
[2]
The Acute Respiratory Distress Syndrome Network (2003) Effects of recruitment maneuvers in patients with acute lung injury and acute respiratory distress syndrome ventilated with high positive end-expiratory pressure. Crit Care Med 31: 2592–2597.
[3]
The National Heart, Lung, and Blood Institute Acute Respiratory Distress Syndrome (ARDS) Clinical Trials Network (2006) Efficacy and safety of corticosteroids for persistent acute respiratory distress syndrome. N Engl J Med 354: 1671–1684.
[4]
The National Heart, Lung, and Blood Institute Acute Respiratory Distress Syndrome (ARDS) Clinical Trials Network (2006) Comparison of two fluid management strategies in acute lung injury. N Engl J Med 354: 2564–2575.
[5]
Tomashefski JF Jr, Davies P, Boggis C, Greene R, Zapol WM, et al. (1983) The pulmonary vascular lesions of the adult respiratory distress syndrome. Am J Pathol 112: 112–126.
[6]
Orfanos SE, Mavrommati I, Korovesi I, Roussos C (2004) Pulmonary endothelium in acute lung injury: From basic science to the critically ill. Intensive Care Med 30: 1702–1714.
[7]
Yamada M, Kubo H, Kobayashi S, Ishizawa K, Numasaki M, et al. (2004) Bone marrow-derived progenitor cells are important for lung repair after lipopolysaccharide-induced lung injury. J Immunol 172: 1266–1272.
[8]
Yamada M, Kubo H, Ishizawa K, Kobayashi S, Shinkawa M, et al. (2005) Increased circulating endothelial progenitor cells in patients with bacterial pneumonia: evidence that bone marrow derived cells contribute to lung repair. Thorax 60: 410–413.
[9]
Burnham EL, Taylor WR, Quyyumi AA, Rojas M, Brigham KL, et al. (2005) Increased circulating endothelial progenitor cells are associated with survival in acute lung injury. Am J Respir Crit Care Med 172: 854–860.
[10]
Aicher A, Zeiher AM, Dimmeler S (2005) Mobilizing endothelial progenitor cells. Hypertension 45: 321–325.
[11]
Natori T, Sata M, Washida M, Hirata Y, Nagai R, et al. (2002) G-CSF stimulates angiogenesis and promotes tumor growth: potential contribution of bone marrow-derived endothelial progenitor cells. Biochem Biophys Res Commun 297: 1058–1061.
[12]
Aicher A, Heeschen C, Mildner-Rihm C, Urbich C, Ihling C, et al. (2003) Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. Nat Med 9: 1370–1376.
[13]
Duda DG, Fukumura D, Jain RK (2004) Role of eNOS in neovascularization: NO for endothelial progenitor cells. Trends Mol Med 10: 143–145.
[14]
Leone AM, Valgimigli M, Giannico MB, Zaccone V, Perfetti M, et al. (2009) From bone marrow to the arterial wall: the ongoing tale of endothelial progenitor cells. Eur Heart J 30: 890–899.
[15]
Sasaki K, Heeschen C, Aicher A, Ziebart T, Honold J, et al. (2006) Ex vivo pretreatment of bone marrow mononuclear cells with endothelial NO synthase enhancer AVE9488 enhances their functional activity for cell therapy. Proc Natl Acad Sci USA 103: 14537–14541.
[16]
Segal MS, Shah R, Afzal A, Perrault CM, Chang K, et al. (2006) Nitric oxide cytoskeletal-induced alterations reverse the endothelial progenitor cell migratory defect associated with diabetes. Diabetes 55: 102–109.
[17]
Chen YH, Lin SJ, Lin FY, Wu TC, Tsao CR, et al. (2007) High glucose impairs early and late endothelial progenitor cells by modifying nitric oxide-related but not oxidative stress mediated mechanisms. Diabetes 56: 1559–1568.
[18]
Javanmard SH, Gheisari Y, Soleimani M, Nematbakhsh M, Monajemi A (2010) Effect of L-arginine on circulating endothelial progenitor cells in hypercholesterolemic rabbits. Int J Cardiol 143: 213–216.
[19]
Thomassen MJ, Buhrow LT, Connors MJ, Kaneko FT, Erzurum SC, et al. (1997) Nitric oxide inhibits inflammatory cytokine production by human alveolar macrophages. Am J Respir Cell Mol Biol 17: 279–283.
[20]
Tang JR, Markham NE, Lin YJ, McMurtry IF, Maxey A, et al. (2004) Inhaled nitric oxide attenuates pulmonary hypertension and improves lung growth in infant rats after neonatal treatment with a VEGF receptor inhibitor. Am J Physiol Lung Cell Mol Physiol 287: L344–L351.
[21]
Balasubramaniam V, Maxey AM, Morgan DB, Markham NE, Abman SH (2006) Inhaled NO restores lung structure in eNOS-deficient mice recovering from neonatal hypoxia. Am J Physiol Lung Cell Mol Physiol 291: L119–L127.
[22]
Lin YJ, Markham NE, Balasubramaniam V, Tang JR, Maxey A, et al. (2005) Inhaled nitric oxide enhances distal lung growth after exposure to hyperoxia in neonatal rats. Pediatr Res 58: 22–29.
[23]
Tourneux P, Markham N, Seedorf G, Balasubramaniam V, Abman SH (2009) Inhaled nitric oxide improves lung structure and pulmonary hypertension in a model of bleomycin-induced bronchopulmonary dysplasia in neonatal rats. Am J Physiol Lung Cell Mol Physiol 297: L1103–L1111.
[24]
Hu XW, Guo CB, Bo Sun (2007) Inhaled nitric oxide attenuates hyperoxic and inflammatory injury without alteration of phosphatidylcholine synthesis in rat lungs. Pulm Pharmacol Ther 20: 75–84.
[25]
Song J, Palmer K, Sun B (2010) Effects of inhaled nitric oxide and surfactant with extracorporeal life support in recovery phase of septic acute lung injury in piglets. Pulm Pharmacol Ther 23: 78–87.
[26]
Zhu YR, Guo CB, Cao L, Gong XH, Wang CQ, et al. (2005) Different effects of surfactant and inhaled nitric oxide in modulation of inflammatory injury in ventilated piglet lungs. Pulm Pharmacol Ther 18: 303–313.
[27]
Qian L, Liu H, Yu W, Wang X, Sun Z, et al. (2008) Effects of positive end-expiratory pressure, inhaled nitric oxide and surfactant on expression of proinflammatory cytokines and growth factors in preterm piglet lungs. Pediatr Res 64: 17–23.
[28]
Fadini GP, Sartore S, Albiero M, Baesso I, Murphy E, et al. (2006) Number and function of endothelial progenitor cells as a marker of severity for diabetic vasculopathy. Arterioscler Thromb Vasc Biol 26: 2140–2146.
[29]
Kondo T, Hayashi M, Takeshita K, Numaguchi Y, Kobayashi K, et al. (2004) Smoking cessation rapidly increases circulating progenitor cells in peripheral blood in chronic smokers. Arterioscler Thromb Vasc Biol 24: 1442–1447.
[30]
Zhou ZH, Sun B, Lin K, Zhu LW (2000) Prevention of rabbit acute lung injury by surfactant, inhaled nitric oxide, and pressure support ventilation. Am J Respir Crit Care Med 161: 581–588.
[31]
Hawkes SP, Li H, Taniguchi GT (2001) Zymography and reverse zymography for detecting MMPs, and TIMPs. Clark II (Ed.), Matrix Metalloproteinases Protocols. Humana Press 399–410.
[32]
Timmermans F, Plum J, Yoder MC, Ingram DA, Vandekerckhove B, et al. (2009) Endothelial progenitor cells: identity defined? J Cell Mol Med 13: 87–102.
[33]
Case J, Mead LE, Bessler WK, Prater D, White HA, et al. (2007) Human CD341AC1331VEGFR-2 cells are not endothelial progenitor cells but distinct, primitive hematopoietic progenitors. Exp Hematol 35: 1109–1118.
[34]
Kheirandish-Gozal L, Bhattacharjee R, Kim J, Clair HB, Gozal D (2010) Endothelial progenitor cells and vascular dysfunction in children with obstructive sleep apnea. Am J Respir Crit Care Med 182: 92–97.
[35]
Padfield GJ, Tura O, Haeck ML, Short A, Freyer E, et al. (2010) Circulating endothelial progenitor cells are not affected by acute systemic inflammation. Am J Physiol Heart Circ Physiol 298: H2054–2061.
[36]
Lam CF, Liu YC, Hsu JK, Yeh PA, Su TY, et al. (2008) Autologous Transplantation of Endothelial Progenitor Cells Attenuates Acute Lung Injury in Rabbits. Anesthesiology 108: 392–401.
[37]
K？hler CM, Wechselberger J, Hilbe W, Gschwendtner A, Colleselli D, et al. (2007) Peripheral infusion of rat bone marrow derived endothelial progenitor cells leads to homing in acute lung injury. Respir Res 8: 50.
[38]
Balasubramaniam V, Ryan SL, Seedorf GJ, Roth EV, Heumann TR, et al. (2010) Bone marrow-derived angiogenic cells restore lung alveolar and vascular structure after neonatal hyperoxia in infant mice. Am J Physiol Lung Cell Mol Physiol 298: L315–L323.
[39]
Pitchford SC, Furze RC, Jones CP, Wengner AM, Rankin SM (2009) Differential Mobilization of Subsets of Progenitor Cells from the Bone Marrow. Cell Stem Cell 4: 62–72.
[40]
Fukumura D, Gohongi T, Kadambi A, Izumi Y, Ang J, et al. (2001) Predominant role of endothelial nitric oxide synthase in vascular endothelial growth factor-induced angiogenesis and vascular permeability. Proc Natl Acad Sci USA 98: 2604–2609.
[41]
Kong D, Melo LG, Gnecchi M, Zhang L, Mostoslavsky G, et al. (2004) Cytokine-induced mobilization of circulating endothelial progenitor cells enhances repair of injured arteries. Circulation 110: 2039–2046.
[42]
Dzau VJ, Gnecchi M, Pachori AS, Morello F, Melo LG (2005) Therapeutic potential of endothelial progenitor cells in cardiovascular diseases. Hypertension 46: 7–18.
[43]
Balasubramaniam V, Mervis CF, Maxey AM, Markham NE, Abman SH (2007) Hyperoxia reduces bone marrow, circulating, and lung endothelial progenitor cells in the developing lung: implications for the pathogenesis of bronchopulmonary dysplasia. Am J Physiol Lung Cell Mol Physiol 292: L1073–L1084.
[44]
Voelkel NF, Vandivier RW, Tuder RM (2006) Vascular endothelial growth factor in the lung. Am J Physiol Lung Cell Mol Physiol 290: L209–L221.
[45]
Lopez E, Boucherat O, Franco-Montoya ML, Bourbon JR, Delacourt C, et al. (2006) Nitric oxide donor restores lung growth factor and receptor expression in hyperoxia-exposed rat pups. Am J Respir Cell Mol Biol 34: 738–745.
