SN50, a Cell-Permeable Inhibitor of Nuclear Factor-κB, Attenuates Ventilator-Induced Lung Injury in an Isolated and Perfused Rat Lung Model.

High tidal volume (VT) ventilation causes the release of various mediators and results in ventilator-induced lung injury (VILI). SN50, a cell-permeable nuclear factor-κB (NF-κB) inhibitory peptide, attenuates inflammation and acute respiratory distress syndrome. However, the mechanisms associated with the effects of SN50 in VILI have not been fully elucidated. We investigated the cellular and molecular mechanisms for the effects of SN50 treatment in VILI. An isolated and perfused rat lung model was exposed to low (5 mL/kg) or high (15 mL/kg) VT ventilation for 6 h. SN50 was administered in the perfusate at the onset of the high-stretch mechanical ventilation. The hemodynamics, lung histological changes, inflammatory responses, and activation of apoptotic pathways were evaluated. VILI was demonstrated by increased pulmonary vascular permeability and lung weight gain, as well as by increased levels of interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), hydrogen peroxide, and macrophage inflammatory protein-2 in the bronchoalveolar lavage fluid. The lung tissue expression of TNF-α, IL-1ß, mitogen-activated protein kinases (MAPKs), caspase-3, and phosphorylation of serine/threonine-specific protein kinase (p-AKT) was greater in the high VT group than in the low VT group. Upregulation and activation of NF-κB was associated with increased lung injury in VILI. SN50 attenuated the inflammatory responses, including the expression of IL-1ß, TNF-α, MPO, MAPKs, and NF-κB. In addition, the downregulation of apoptosis was evaluated using caspase-3 and p-AKT expression. Furthermore, SN50 mitigated the increases in the lung weights, pulmonary vascular permeability, and lung injury. In conclusion, VILI is associated with inflammatory responses and activation of NF-κB. SN50 inhibits the activation of NF-κB and attenuates VILI.

Spillover of cytokines and reactive oxygen species in ventilator-induced lung injury associated with inflammation and apoptosis in distal organs.

BACKGROUND: The mechanism between ventilator-induced lung injury (VILI) and multiple organ injury is unclear. The aim of our study was to investigate the mechanisms of VILI-induced distal organ injury. METHODS: VILI was induced in rat lungs with high tidal volume (V(T)) ventilation of 40 mL/kg for 6 h. Rats with low V(T) ventilation of 6 mL/kg served as controls. Inflammatory and apoptotic indices in lung and distal organs were assessed. RESULTS: VILI increased lung weight, airway pressure, inflammation, and apoptotic pathologic changes without hemodynamic changes. The white blood cell count and the levels of H2O2, interleukin-1ß (IL-1ß), tumor necrosis factor alpha, and macrophage inflammatory protein-2 in bronchoalveolar lavage fluid were higher in the VILI group compared with the control group. H2O2, IL-1ß, and tumor necrosis factor alpha in blood from the left ventricle were up-regulated. H2O2, IL-1ß, tumor necrosis factor alpha, macrophage inflammatory protein-2, c-Jun N-terminal kinase, p38, nuclear factor kappa B, and caspase-3 in lung, heart, liver, and kidney tissues in the VILI group were up-regulated. Furthermore, the apoptotic score for the kidneys was higher than those for other distal organs in the VILI group. CONCLUSIONS: High V(T) ventilation induces VILI and is associated with inflammation and apoptosis in distal organs. Up-regulation of reactive oxygen species and cytokines in VILI is associated with systemic inflammatory responses. Kidney tissue appears to be more vulnerable than heart and liver tissues following VILI.

Inhibitor of nuclear factor-κB, SN50, attenuates lipopolysaccharide-induced lung injury in an isolated and perfused rat lung model.

NF-κB cell permeable inhibitory peptide (SN50) inhibits translocation of nuclear factor-κB (NF-κB) and production of inflammatory cytokines that are implicated in lipopolysaccharide (LPS)-induced lung injury (LPSLI). However, the protective effect of SN50 in LPSLI is unclear. We explored the cellular and molecular mechanisms of SN50 treatment in LPSLI. LPSLI was induced by intratracheal instillation of 10 mg/kg LPS using an isolated and perfused rat lung model. SN50 was administered in the perfusate 15 minutes before LPS was administered. Hemodynamics, lung histologic change, inflammatory responses, and activation of apoptotic pathways were evaluated. After LPSLI, increased pulmonary vascular permeability and lung weight gain was observed. The levels of interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, myeloperoxidase, and macrophage inflammatory protein-2 increased in bronchoalveolar lavage fluids. Lung-tissue expression of TNF-α, IL-1ß, mitogen-activated protein kinases (MAPKs), caspase-3, p-AKT (serine-threonine kinase, also known as protein kinase B), and plasminogen activator inhibitor-1 (PAI-1) was greater in the LPS group compared with controls. Upregulation and activation of NF-κB was associated with increased lung injury in LPSLI. SN50 attenuated the inflammatory responses, including expression of IL-1ß, TNF-α, myeloperoxidase, MAPKs, PAI-1, and NF-κB; downregulation of apoptosis indicated by caspase-3 and p-AKT expression was also observed. In addition, SN50 mitigated the increase in the lung weight, pulmonary vascular permeability, and lung injury. In conclusion, LPSLI is associated with inflammatory responses, apoptosis, and coagulation. NF-κB is an important therapeutic target in the treatment of LPSLI. SN50 inhibits translocation of NF-κB and attenuates LPSLI.

Transforming growth factor-ß1 and tumor necrosis factor-α are associated with clinical severity and airflow limitation of COPD in an additive manner.

BACKGROUND: The role of tumor necrosis factor-α (TNF-α) and transforming growth factor-ß1 (TGF-ß1) in chronic obstructive pulmonary disease (COPD) is controversial. The purpose of this study was to assess the relationships among polymorphisms, clinical phenotypes, and the serum levels of TNF-α and TGF-ß1. METHODS: Polymorphisms of promoters of TNF-α (rs 361525 and rs 1800629) and TGF-ß1 (rs 1800469) in 110 COPD patients, 110 nonsmoker health controls without COPD, and 34 smokers were evaluated. Pulmonary functions, chest computed tomography, TGF-ß1, and TNF-α were assessed. RESULTS: The genetic polymorphism of TNF-α (rs 361525) was associated with COPD. More severe COPD patients had higher serum levels of TNF-α and TGF-ß1; moreover, serum levels of TGF-ß1of mild COPD patients were higher than normal controls. All of the studied subjects were divided into four groups by the 95th percentile value of control as cutoff serum value of TGF-ß1 (224.35 ρg/ml) or TNF-α (17.56 ρg/ml) to define the high value of TGF-ß1 or TNF-α, which are higher than those cutoff of values (>224.35 or 17.56 ρg/ml). The FEV1 of the group with high TGF-ß1 + low TNF-α or low TGF-ß1 + high TNF-α or high TNF-α + high TGF-ß1 was lower than the group with low TGF-ß1 + low TNF-α group. Moreover, the lowest value of FEV1 was in the group with high TNF-α + high TGF-ß1. CONCLUSIONS: The genetic polymorphism of the TNF-α is associated with COPD. Both TGF-ß1 and TNF-α modulate clinical severity and airflow limitation in an additive manner.

Effects of guideline-oriented pharmacotherapy in patients with newly diagnosed COPD: a prospective study.

BACKGROUND: Whether guideline-oriented pharmacotherapy prevents the decline in pulmonary function or reduces systemic inflammation associated with chronic obstructive pulmonary disease (COPD) is uncertain. OBJECTIVES: The aim of this study was to assess the outcome of COPD in clinical practice under real-world conditions in Taiwan as measured by pulmonary function and systemic inflammation parameters (C-reactive protein (CRP) or white blood cell (WBC)) after initiation of guideline-oriented pharmacotherapy. METHODS: Newly diagnosed COPD patients were enrolled and prospectively observed in real-world outpatient practice following initiation of pharmacotherapy of COPD. Pulmonary function, WBC and neutrophil counts, and CRP level of COPD patients were assessed annually. This study enrolled 566 patients and 263 returned for follow-up visits. RESULTS: Significantly higher postbronchodilator FVC, FEV1, and FEV1/FVC but lower DLCO were found at 1 year compared to baseline values. During 4-year follow-up period, FVC and FEV1 remained stable. DLCO progressively declined compared to baseline. No significant changes were seen in CRP and neutrophil count over a 3-year period. Values of CRP, WBC, and neutrophil count correlated inversely with FEV1, FVC, FEV1/FVC, and DLCO. CONCLUSIONS: Guideline-oriented pharmacotherapy of COPD improves airflow limitation but does not prevent the alveolar destruction and systemic inflammation under real-world conditions in Taiwan.

Genetic polymorphism of transforming growth factor ß1 and tumor necrosis factor α is associated with asthma and modulates the severity of asthma.

BACKGROUND: The role of transforming growth factor ß1 (TGF-ß1) and tumor necrosis factor α (TNF-α) in asthma is unclear. The aim of this study was to assess the relationships among polymorphisms, clinical phenotypes, and the serum levels of TGF-ß1 and TNF-α. METHODS: Polymorphisms of promoter of TGF-ß1 (C-509T locus) and TNF-α (G-308 A locus; rs 1800629) in 217 asthmatic patients and 110 healthy controls were evaluated. Pulmonary function, total immunoglobulin E (IgE), specific IgE antibodies, total eosinophil counts, TGF-ß1, and TNF-α were assessed. RESULTS: The genetic polymorphisms of TGF-ß1 promoter and TNF-α were significantly associated with asthma. Subjects with more severe asthma had higher serum levels of TGF-ß1 and TNF-α. In asthmatic subjects the TGF-ß1 of atopic subjects was higher than those without atopy. All studied subjects (asthma plus control) were divided into 4 groups by mean value of TGF-ß1 or TNF-α. The high values of TGF-ß1 or TNF-α were defined by higher than the mean values of the studied subjects of TGF-ß1 (392.42 pg/mL) and TNF-α (55.86 pg/mL). The FEV1 of the group with high TGF-ß1 plus low TNF-α was lower than that in the group with low TGF-ß1 plus low TNF-α. The lowest FEV1 was in the group with high TNF-α and high TGF-ß1. CONCLUSIONS: The genetic polymorphisms of TGF-ß1 and TNF-α are associated with asthma. TGF-ß1 modulates atopy. Both TGF-ß1 and TNF-α modulate clinical severity and airway obstruction, in an additive manner.

Apocynin attenuates lipopolysaccharide-induced lung injury in an isolated and perfused rat lung model.

Apocynin (Apo) suppresses the generation of reactive oxygen species that are implicated in lipopolysaccharide (LPS)-induced lung injury (LPSLI). We thus hypothesized that Apo may attenuate LPSLI. In addition, we explored the cellular and molecular mechanisms of Apo treatment in LPSLI. Lipopolysaccharide-induced lung injury was induced by intratracheal instillation of 10 mg/kg LPS in isolated and perfused rat lung model. Apocynin was administered in the perfusate at 15 min before LPS was administered. Hemodynamics, lung injury indices, inflammatory responses, and activation of apoptotic pathways were assessed. There was an increase in lung vascular permeability associated with lung weight gain after LPS exposure. The levels of interleukin 1ß (IL-1ß), tumor necrosis factor α (TNF-α), macrophage inflammatory protein 2, H2O2, and albumin increased in the bronchoalveolar lavage fluid. Adhesion molecule of neutrophil (CD31) was upregulated. The expression of TNF-α, IL-1ß, glutathione, myeloperoxidase, JNK, P38, caspase 3, p-AKT, and plasminogen activator inhibitor 1 in lung tissue was greater in the LPS groups when compared with the control group. Upregulation and activation of nuclear factor κB occurred along with increased histopathologic lung injury score in LPSLI. The Apo attenuated these inflammatory responses including the levels of CD31, H2O2, TNF-α, IL-1ß, myeloperoxidase, P38, and nuclear factor κB along with downregulation of apoptosis as reflected by caspase 3 and p-AKT. In addition, Apo attenuated the increase in lung weight, bronchoalveolar lavage fluid albumin content, and the histopathologic lung injury score. In conclusion, LPSLI is associated with increased inflammatory responses, apoptosis, and coagulation. The administration of Apo attenuates LPSLI through downregulation of the inflammatory responses and apoptosis.

N-acetylcysteine attenuates ventilator-induced lung injury in an isolated and perfused rat lung model.

N-acetylcysteine (NAC) suppresses the generation of reactive oxygen species (ROS) that are implicated in ventilator-induced lung injury (VILI). We thus hypothesised that NAC attenuates VILI. VILI was induced by mechanical ventilation with a tidal volume (Vt) of 15mlkg(-1) in isolated and perfused rat lung. NAC was administered in the perfusate prior to the onset of mechanical ventilation. A group ventilated with low Vt of 5mlkg(-1) served as control. Haemodynamics, lung injury indices, inflammatory responses and activation of apoptotic pathways were determined upon completion of the mechanical ventilation. There was an increase in lung permeability and lung weight gain after mechanical ventilation with high Vt, compared to low Vt. The levels of inflammatory cytokines including interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α) and macrophage inflammatory protein-2 (MIP-2) increased in lung lavage fluids; the concentrations of H(2)O(2) were higher in lung lavage fluids, and the expression of myeloperoxidase (MPO), JNK, P38, pAKT and caspase-3 in lung tissue was greater in the high Vt than in the low Vt group. The concentrations of glutathione (GSH) in lung tissue were higher in low Vt than those in high Vt. The administration of NAC increased GSH, attenuated ROS, cytokines, MPO, JNK, pAKT and caspase-3 and lung permeability associated with decreased activation of nuclear factor-κB. VILI is associated with inflammatory responses including the generation of ROS, cytokines and the activation of mitogen-activated protein kinase cascade. The administration of NAC attenuates the inflammatory responses, apoptosis and VILI in the isolated, perfused rat lung model.

Apocynin attenuates ischemia-reperfusion lung injury in an isolated and perfused rat lung model.

Apocynin suppresses the generation of reactive oxygen species (ROS) that are implicated in ischemia-reperfusion (I/R) lung injury. We thus hypothesized that apocynin attenuates I/R. Furthermore, we explored the mechanisms by which apocynin may attenuate I/R. I/R was induced in an isolated and perfused rat lung model with ischemia for 1 h followed by reperfusion for 1 h. Apocynin was administered in the circulating perfusate at the onset of ischemia. Hemodynamics, lung injury indices, inflammatory responses, and activation of apoptotic pathways were determined. An increase in lung permeability and lung weight gain was noted after I/R. Peak airway pressure was increased, and pH of circulating perfusate was decreased. The adhesion molecule of neutrophil (CD31) in perfusate was upregulated. The levels of albumin, white blood cell count, and inflammatory cytokines including interleukin-1ß, tumor necrosis factor-α, and macrophage inflammatory protein-2 increased in lung lavage fluid; the concentrations of carbonyl and thiobarbituric acid reactive substances were greater in the circulating perfusate; and the expression of myeloperoxidase, JNK, P38, and caspase-3 in lung tissue was greater in the control group. Upregulation and activation of nuclear factor-κB (NF-κB) in nuclei were found in I/R. The administration of apocynin attenuated these inflammatory responses and lung permeability associated with decreased activation of NF-κB. We conclude that I/R is associated with inflammatory responses including the generation of ROS, adhesion protein of neutrophil, cytokines, and the activation of mitogen-activated protein kinase and NF-κB cascade. The administration of apocynin attenuates the inflammatory responses and I/R in the isolated, perfused rat lung model.

Apocynin attenuates ventilator-induced lung injury in an isolated and perfused rat lung model.

RATIONALE: Apocynin suppresses the generation of reactive oxygen species (ROS) that are implicated in ventilator-induced lung injury (VILI). We thus hypothesized that apocynin attenuates VILI. METHODS: VILI was induced by mechanical ventilation with tidal volume (V(t)) of 15 ml/kg in isolated and perfused rat lung. Apocynin was administered in the perfusate at onset of mechanical ventilation. A group ventilated with low V(t) of 5 ml/kg served as control. Hemodynamics, lung injury indices, inflammatory responses, and activation of apoptotic pathways were determined upon completion of mechanical ventilation. RESULTS: There was an increase in lung permeability and lung weight gain after mechanical ventilation with high V(t), compared with low V (t). Levels of inflammatory cytokines including interleukin-1ß (IL-1ß), tumor necrosis factor-alpha (TNF-α), and macrophage inflammatory protein-2 (MIP-2) increased in lung lavage fluids; concentrations of carbonyl, thiobarbituric acid reactive substances, and H(2)O(2) were higher in perfusates and lung lavage fluids, and expression of myeloperoxidase, JNK, p38, and caspase-3 in lung tissue was greater in the high-V(t) than in the low-V(t) group. Administration of apocynin attenuated these inflammatory responses and lung permeability associated with decreased activation of nuclear factor-κB. CONCLUSIONS: VILI is associated with inflammatory responses including generation of ROS, cytokines, and activation of mitogen-activated protein kinase cascades. Administration of apocynin at onset of mechanical ventilation attenuates inflammatory responses and VILI in the isolated, perfused rat lung model.

Ventilator-induced lung injury (VILI) promotes ischemia/reperfusion lung injury (I/R) and NF-kappaB antibody attenuates both injuries.

RATIONALE: Whether the ventilator-induced lung injury (VILI) superimposed on ischemia/reperfusion injury (I/R) causes synergistic damage has not been well explored. Whether nuclear factor-kappa B (NF-kappaB) antibody has protective effects for both injuries is also unknown. METHODS: I/R and VILI were produced in an isolated rat lung model. Hemodynamics, lung weight gain (LWG), capillary filtration coefficient (K(fc)), cytokines, and lung pathology were assessed. RESULTS: VILI or I/R produced similar permeability pulmonary edema which was reflected by increasing K(fc) and LWG. Cytokine (IL-1beta) up-regulation occurred in both injuries. Pathologic examination showed edema and inflammatory cell infiltration in VILI or I/R. In addition, the alveoli were overdistended and even ruptured because of marked inhomogeneity of inflation in VILI. Furthermore, combined I/R and VILI produced further increases in K(fc), LWG, IL-1beta, as well as more severe pathologic changes. Conversely, less permeability pulmonary edema, pathologic changes and IL-1 expression were found in groups pretreated with anti-NF-kappaB antibody. CONCLUSION: VILI and I/R cause synergistic damage on the lung. I/R or VILI alone or combined can be attenuated by NF-kappaB antibody. NF-kappaB plays an important role in both forms of lung injury. We propose anti-NF-kappaB antibody pretreatment to be beneficial for VILI, I/R and lung transplantation.