Expression of Inducible Nitric Oxide Synthase and Inflammatory Cytokines in Alveolar Macrophages of ARDS Following Sepsis

Kobayashi, Atsuko; Hashimoto, Satoru; Kooguchi, Kunihiko; Kitamura, Yoshihiro; Onodera, Hidetoshi; Urata, Yoji; Ashihara, Tsukasa

doi:10.1378/chest.113.6.1632

Cited by 102 publications

(53 citation statements)

References 29 publications

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“…We found an increased iNOS mRNA expression in cells that were isolated from urine of both septic patients and healthy volunteers who were treated with endotoxin. Until now, expression of iNOS in humans has been shown only in alveolar macrophages from patients with acute respiratory distress syndrome (32) and in mesenteric arterial smooth muscle and peripheral blood mononuclear cells (33) after sepsis. Another recent study did not detect iNOS in cells or vessels of the systemic circulation in patients with septic shock that was caused by cellulitis but only at the site of infection (34).…”

Section: Discussionmentioning

confidence: 99%

Upregulation of Renal Inducible Nitric Oxide Synthase during Human Endotoxemia and Sepsis Is Associated with Proximal Tubule Injury

Heemskerk

Pickkers

Bouw

et al. 2006

Clinical Journal of the American Society of Nephrology

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The incidence and the mortality of septic acute kidney injury are high, partly because the pathogenesis of sepsis-induced renal dysfunction is not clear. The objective of this study was to investigate the upregulation of renal inducible nitric oxide synthase (iNOS) in human endotoxemia and sepsis and the effect of NO on tubular integrity. Septic patients and endotoxemia that was induced by a bolus injection of 2 ng/kg Escherichia coli LPS in human volunteers were studied. In addition, the effect of co-administration of the selective iNOS inhibitor aminoguanidine was evaluated. The urinary excretion of the cytosolic glutathione-S-transferase-A1 (GSTA1-1) and GSTP1-1, markers for proximal and distal tubule damage, respectively, was determined. In septic patients, an almost 40-fold induction of iNOS mRNA in cells that were isolated from urine was found accompanied by a significant increase in NO metabolites in blood. The mRNA expression of iNOS was induced 34-fold after endotoxin administration. LPS-treated healthy volunteers showed a higher urinary excretion of NO metabolites compared with control subjects. Urinary NO metabolite excretion correlated with urinary GSTA1-1 excretion, indicating proximal tubule damage, whereas no distal tubular damage was observed. Co-administration of aminoguanidine reduced the upregulation of iNOS mRNA, urinary NO metabolite, and GSTA1-1 excretion, indicating that upregulation of iNOS and subsequent NO production may be responsible for renal proximal tubule damage observed.

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Section: Discussionmentioning

confidence: 99%

Upregulation of Renal Inducible Nitric Oxide Synthase during Human Endotoxemia and Sepsis Is Associated with Proximal Tubule Injury

Heemskerk

Pickkers

Bouw

et al. 2006

Clinical Journal of the American Society of Nephrology

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“…There is growing evidence that certain forms of acute and chronic pulmonary inflammation such as bronchial asthma [22], ARDS [5], IPF [3,9], tuberculosis [1], and sarcoidosis [2] are associated with the upregulation of pulmonary NOS2 expression. In addition, an increase of NO in exhaled air is measured in these diseases.…”

Section: Discussionmentioning

confidence: 99%

“…Nitric oxide (NO) is a gas that can be detected in exhaled air and its increase is observed in several pulmonary disorders including tuberculosis, sarcoidosis, idiopathic pulmonary fibrosis (IPF), primary lung cancer, and adult respiratory distress syndrome (ARDS) [1][2][3][4][5]. Exhaled NO has been shown to be an indirect marker of airway inflammation; alveolar macrophages (AM) and airway epithelial cells are assumed to be the main source of the elevated levels of NO [1][2][3][4][5].…”

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confidence: 99%

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Human alveolar epithelial cells induce nitric oxide synthase‐2 expression in alveolar macrophages

Pechkovsky¹,

Zissel²,

Stamme³

et al. 2002

Eur Respir J

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It was hypothesized that cell-to-cell interaction between human alveolar macrophages (AM) and alveolar epithelium, might be an important factor leading to nitric oxide synthase-2 (NOS2) messenger ribonucleic acid (mRNA) and protein expression by constituent cells of the alveolar wall and/or AM.NOS2 mRNA and the protein expression patterns of human AM and alveolar epithelial cells type II (AEC-II) isolated from normal parts of lung resections of patients with pulmonary malignancies were determined. In addition, NOS2 mRNA expression in human AM co-cultured with autologous AEC-II in the presence of pro-inflammatory cytokines interleukin (IL)-1b, tumour necrosis factor (TNF)-a, interferon (IFN)-c or lipopolysaccharide (LPS) was investigated. The effect of human surfactant protein-A (SP-A) on IFN-c-mediated NOS2 mRNA expression in human AM was also studied.Neither NOS2 mRNA nor protein could be detected in freshly isolated, unstimulated or cytokine-stimulated AEC-II. In contrast, freshly isolated AM from bronchoalveolar lavage or lung tissue samples expressed immunoreactivity for NOS2 protein, but no NOS2 mRNA could be detected by reverse transcriptase polymerase chain reaction. All stimuli tested failed to induce NOS2 mRNA expression in human AM in vitro. Only AM-AEC-II co-culture in the presence of IFN-c led to NOS2 mRNA and protein expression. In situ hybridization of NOS2 mRNA on lung tissue explants and immunohistochemical staining of cytospin preparations of AM-AEC-II co-cultures demonstrated that NOS2 is expressed in AM but not in AEC-II. This co-culture effect could not be reproduced by substitution of AEC-II with SP-A.These data give evidence of a regulatory network controlling human nitric oxide synthase-2 expression in the lower respiratory tract.

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“…It may also be possible that the stimuli and culture conditions that we used are inadequate to stimulate human AEC II to express NOS2 mRNA and, therefore, do not represent the complex dynamic processes occurring in vivo, especially during lung inflammation. Although several reports concerning NOS2 expression in normal human lung tissue have provided evidence that peripheral lung parenchyma, as well as resident lung macrophages, do not express NOS2 mRNA (10) and that no specific immunostaining for NOS2 protein can be observed in alveolar compartments (18), the accumulation of NOS2 protein and NO production have been documented in AM isolated from patients with active pulmonary tuberculosis (22) and acute respiratory distress syndrome (17). However, many attempts to induce NOS2 mRNA expression and/or NO production by human AM in vitro failed (5,7,10).…”

Section: Discussionmentioning

confidence: 99%

Pattern of NOS2 and NOS3 mRNA expression in human A549 cells and primary cultured AEC II

Pechkovsky

Zissel

Goldmann

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

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The human alveolar type II epithelium-like cell line A549 expresses nitric oxide synthase type 2 (NOS2), but not NOS3, and produces nitric oxide (NO) upon appropriate stimulation. However, relatively little is known regarding the NOS2 and NOS3 expression of type II human alveolar epithelial cells (AEC II) in primary culture. We detected NOS3 mRNA in freshly isolated AEC II and after 24 h of culture. NOS3 mRNA levels were much higher in AEC II cultured for 24 h with or without interferon-␥, interleukin-1␤, and tumor necrosis factor-␣, compared with freshly isolated cells. Cytokine stimulation did not change the NOS3 mRNA expression level in AEC II compared with unstimulated cells. NOS3 protein expression was verified by Western blot, and measuring nitrate/nitrite revealed that the protein is active. In contrast, neither NOS2 mRNA nor protein could be detected in freshly isolated, unstimulated or cytokine-stimulated human AEC II in 24-or 72-h primary cultures, whereas A549 cells expressed NOS2 message and protein upon stimulation with proinflammatory cytokines. In situ hybridization confirmed that AEC II express NOS3, but not NOS2 mRNA in vivo. These data demonstrate that there are significant differences between primary AEC II and A549 cells in NOS mRNA expression pattern.nitric oxide synthase; mRNA; RT-PCR; interferon-␥; tumor necrosis factor-␣; interleukin-1␤; type II human alveolar epithelial cells; A549 cell line NITRIC OXIDE SYNTHASES (NOS) constitute a family with at least three distinct isoforms; neuronal (nNOS or NOS1), inducible (iNOS or NOS2) and endothelial (eNOS or NOS3) (20, 21). NOS1 and NOS3 are constitutively active and produce small amounts of nitric oxide (NO), whereas NOS2 is induced by inflammatory cytokines and produces a larger amount of NO. All three types of NOS are expressed in human lung, and NOS mRNA expression and enzyme activity have been described in human bronchial epithelial cells, macrophages, endothelial cells, and vascular smooth muscle cells (18,34). However, relatively little is known regarding the expression of NOS2 and NOS3 in type II human alveolar epithelial cells (AEC II) and how these enzymes are regulated. Although several laboratories have reported that rat AEC II express NOS2 mRNA and produce NO in vitro (12,26) and that human alveolar epithelium exhibits an NOS-like enzymatic activity in vivo (18), there has been neither molecular nor biochemical confirmation of the expression of NOS2 and NOS3 in human primary cultured AEC II. Asano et al. (4) and Kwon and George (19) have demonstrated that the human type II alveolar epithelial cell line (A549) expresses NOS2 and produces NO in response to proinflammatory cytokines such as interferon-␥ (IFN-␥), interleukin-1␤ (IL-1␤), and tumor necrosis factor-␣ (TNF-␣). Therefore, the aims of the present study were 1) to determine whether human AEC II express NOS2 and NOS3 mRNA in primary culture, 2) to characterize the regulation of mRNA expression of NOS isoforms in these cells by proinflammatory cytokines, and 3) to compare these...

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Expression of Inducible Nitric Oxide Synthase and Inflammatory Cytokines in Alveolar Macrophages of ARDS Following Sepsis

Cited by 102 publications

References 29 publications

Upregulation of Renal Inducible Nitric Oxide Synthase during Human Endotoxemia and Sepsis Is Associated with Proximal Tubule Injury

Upregulation of Renal Inducible Nitric Oxide Synthase during Human Endotoxemia and Sepsis Is Associated with Proximal Tubule Injury

Human alveolar epithelial cells induce nitric oxide synthase‐2 expression in alveolar macrophages

Pattern of NOS2 and NOS3 mRNA expression in human A549 cells and primary cultured AEC II

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