help button home button
AJRCCM
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Rahman, I.
Right arrow Articles by De Boer, W. I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rahman, I.
Right arrow Articles by De Boer, W. I.
American Journal of Respiratory and Critical Care Medicine Vol 166. pp. 490-495, (2002)
© 2002 American Thoracic Society

Original Article

4-Hydroxy-2-Nonenal, a Specific Lipid Peroxidation Product, Is Elevated in Lungs of Patients with Chronic Obstructive Pulmonary Disease

Irfan Rahman, Annemarie A. M. van Schadewijk, Ann J. L. Crowther, Pieter S. Hiemstra, Jan Stolk, William MacNee and Willem I. De Boer

Respiratory Medicine Unit, ELEGI Laboratory, University of Edinburgh Medical School, Edinburgh, United Kingdom; Department of Pulmonology, Leiden University Medical Centre, Leiden; and Department of Pulmonology, Erasmus University, Rotterdam, The Netherlands

Correspondence and requests for reprints should be addressed to Dr. I. Rahman, ELEGI Laboratory, Respiratory Medicine Unit, University of Edinburgh Medical School, Wilkie Building, Teviot Place, Edinburgh EH8 9AG, UK. E-mail: irfan.rahman{at}ed.ac.uk


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Cigarette smoking results in oxidative stress and inflammation in the lungs, which are involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). 4-Hydroxy-2-nonenal (4-HNE), a highly reactive diffusible product of lipid peroxidation, is a key mediator of oxidant-induced cell signaling and apoptosis. 4-HNE has a high affinity toward cysteine, histidine, and lysine groups and forms direct protein adducts. We investigated the presence of 4-HNE–modified proteins in lung tissue obtained from subjects with and without COPD. We studied 23 current or ex-smokers with similar smoking histories with COPD (n = 11; FEV1 < 70% predicted) or without COPD (n = 12; FEV1 > 84% predicted) who had undergone lung resection. As 4-HNE and transforming growth factor-ß1 (TGF-ß1) can modulate {gamma}-glutamylcysteine synthetase ({gamma}-GCS) mRNA levels in lung cells, we assessed the relations between 4-HNE–modified protein levels, FEV1, {gamma}-GCS, and TGF-ß1. 4-HNE–modified protein levels were elevated in airway and alveolar epithelial cells, endothelial cells, and neutrophils in subjects with COPD, compared with the levels in subjects without COPD (p < 0.01). We also observed a significant inverse correlation between the levels of 4-HNE adducts in alveolar epithelium, airway endothelium, and neutrophils and FEV1 (p < 0.05) and a positive correlation between 4-HNE adducts and TGF-ß1 protein and mRNA as well as {gamma}-GCS mRNA levels in airway and alveolar epithelium (p < 0.01). The elevated levels of 4-HNE may play a role in the signaling events in lung inflammation leading to the imbalance of the expression of both proinflammatory mediators and protective antioxidant genes in COPD.

Key Words: 4-hydroxy-2-nonenal • glutathione • transforming growth factor-ß1 • lungs • chronic obstructive pulmonary disease


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Chronic obstructive pulmonary disease (COPD) is a condition characterized by airway inflammation and progressive and largely irreversible airway obstruction (1–3). Cigarette smoking is a major risk factor for COPD (1, 2). However, only 15 to 20% of cigarette smokers appear to be susceptible to its effects and show a rapid decline in FEV1 and develop the disease (1). The reason why only some cigarette smokers are susceptible is unclear at present but may relate to differences between their responses to cigarette smoke. The creation of an imbalance between oxidants and antioxidants (oxidative stress) is considered to be an important event in the pathogenesis of COPD (4). This may be related to a "susceptibility" to the oxidative effects of cigarette smoke and hence to the inflammatory response that ensues.

The sources of the increased oxidative stress in patients with COPD derive from the increased burden of oxidants present in cigarette smoke (5) and from the increased amounts of reactive oxygen species (ROS) released from leukocytes and macrophages, both in the airspaces and in the blood (reviewed in [6–8]). A consequence of oxidative stress is membrane lipid peroxidation in the lungs, primarily involving polyunsaturated fatty acids. The levels of lipid peroxidation products are increased in exhaled air condensate in smokers and in patients with COPD (9–12) (reviewed in [6, 7]). There is increasing evidence that aldehydes, generated endogenously during the process of lipid peroxidation, are involved in many of the pathophysiologic effects associated with oxidative stress in cells and tissues (13). A specific and stable end product of lipid peroxidation, the aldehyde 4-hydroxy-2-nonenal (4-HNE), can diffuse within or even escape from the cell and attack targets far from the site of the original free radical event (14, 15). 4-HNE is a potent alkylating agent that reacts with DNA and proteins, generating various forms of adducts (cysteine, lysine, and histidine residues) (14) that are capable of inducing specific cellular stress responses such as cell signaling and apoptosis (14–16). The lipid aldehyde 4-HNE can be produced from arachidonic acid, linoleic acid, or their hydroperoxides in concentrations of 1 µM to 5 mM, in response to oxidative insults, and is believed to be responsible for many of the effects during oxidative stress in vivo (14). These include transcription of proto-oncogenes, including c-jun, and activation of activator protein 1 (AP-1) via mitogen-activated protein kinase pathways (17, 18). Studies in rat liver epithelial cells have shown that 4-HNE can enter cells and become bound to proteins within the cytosol (15). However, little is known about the localization of 4-HNE in the human lungs.

Our hypothesis is that the interaction of oxidative components of cigarette smoke with cell membrane phospholipids induces alteration in lipid peroxidation products in lung cells in COPD. The degree of formation of the lipid peroxidation product 4-HNE in response to smoking may be a factor in the susceptibility to the development of enhanced airspace inflammation in COPD.

Oxidative stress has been implicated in the expression of both proinflammatory and protective antioxidant genes. Transforming growth factor ß1 (TGF-ß1) is a multifunctional growth factor that modulates cellular proliferation, differentiation, and tissue repair (19). Previously, we have shown an increase in TGF-ß1 expression in bronchiolar and alveolar epithelium in subjects with COPD (20). TGF-ß1 has been shown to decrease glutathione synthesis associated with increased ROS production in human alveolar epithelial cells and pulmonary artery endothelial cells in vitro (21–23). Hence, TGF-ß1 may cause oxidative stress or play a role in membrane lipid peroxidation. In contrast, 4-HNE has been shown to induce expression of the protective antioxidant gene {gamma}-glutamylcysteine synthetase ({gamma}-GCS) mRNA in alveolar epithelial cells (24). We have also shown increased {gamma}-GCS expression in patients with COPD (25). Hence, 4-HNE may play a role in the regulation of TGF-ß1 and {gamma}-GCS gene expression in patients with COPD. We hypothesized that the increased membrane lipid peroxidation (generation of 4-HNE) may be one factor that may provide a signal for the expression of TGF-ß1 and {gamma}-GCS in lungs of patients with COPD. We therefore examined the relationship of levels of 4-HNE to TGF-ß1, {gamma}-GCS, and airflow obstruction in vivo in human lung tissue obtained from subjects with and without COPD. We used immunohistochemistry to investigate the localization and differences in levels of 4-HNE adducts in lungs of subjects with and without COPD. This is the first study to demonstrate the localization of a specific lipid peroxidation molecule in the lungs in inflammatory lung disease.


   METHODS
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Subjects
In this study, we obtained lung tissue specimens from current or ex-smokers with or without COPD who underwent lung resection for lung cancer as described previously (20, 25). Compared with the previous study (20), we included five patients less as the tissue sections were completely used in previous studies. Eleven subjects with COPD (FEV1 < 70% of predicted value before bronchodilatation; seven ex-smokers and four current smokers) and 12 subjects without COPD (FEV1 > 84% predicted; eight ex-smokers and four current smokers) were studied. A summary of the data on lung function tests of these patients is presented in Table 1. The smoking history of the subjects with and without COPD was similar (p > 0.05) (Table 1). The ex-smokers stopped smoking at least 1 year before surgery. All subjects showed less than 13% reversibility of the FEV1 after inhalation of 400 µg of salbutamol. Exclusion criteria included (1) diffuse pulmonary inflammation associated with lung fibrosis, (2) absence of tumor-free or pneumonia-free lung tissue specimens, and (3) obstruction of central bronchi due to the tumor. These exclusion criteria have been used in our previous studies (20, 25, 26). The histologic type of lung cancers was equally distributed in both subject groups. Preoperatively, none of the patients had clinical evidence of an upper respiratory tract infection and none had received antibiotics 4 weeks before operation or glucocorticoids 3 months before operation, with the exception of three patients who received oral glucocorticoids only perioperatively.


View this table:
[in this window]
[in a new window]
  		TABLE 1. A summary of the clinical characteristics of subjects with and without chronic obstructive pulmonary disease*

 
4-HNE Adduct Immunohistochemistry
Lung sections (3-µm thick) were deparaffinized, rehydrated, and pretreated before immunohistochemistry. The sections were refixed, and immunostaining was performed using a mouse monoclonal antibody specific for the 4-HNE adducts (obtained from Japan Institute for the Control of Aging, Fukuroi City, Shizuoka Prefecture, Japan) followed by the avidin–biotin–peroxidase complex (ABC) method, as described by Toyokuni (27). NovaRED (Vector Labs, Burlingame, CA) was used as chromogen. Lung tissue from a patient with adult respiratory distress syndrome was used as a positive control. Omitting the first antibody served as a negative control and resulted in no tissue staining.

The assessment of immunostaining intensity was performed semiquantitatively and in a blinded fashion as described previously (20, 25, 26): 0 = no staining; 1 = weak staining; 2 = moderate staining; and 3 = intense staining. Inflammatory cells were identified by immunostaining for CD68+ cells (for macrophages), CD3 (for T cells), or elastase (for neutrophils). Neutrophils were also identified by the presence of a three-lobed nucleus.

Statistical Analysis
The data are expressed as mean ± SEM. Differences between subject groups were compared by Student's t test. If variances differed, then Welch's correction was applied. Mann–Whitney analysis provided comparable data. For all the data, the distribution was Gaussian. Correlation of the 4-HNE adduct levels with the FEV1, TGF-ß1 mRNA, or {gamma}-GCS mRNA expression was performed using the Pearson correlation test. At p values less than 0.05, differences were considered to be statistically significant.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
In all the subjects, 4-HNE adducts were localized predominantly in the cytoplasm of bronchial, bronchiolar and alveolar epithelial cells, endothelial cells, neutrophils, and CD68+ cells (which are regarded to be macrophages) (Figure 1) . Subepithelial cells (fibroblasts, smooth muscle cells) and lymphocytes (CD3+) were less intensely stained or were not stained. The levels of 4-HNE–modified proteins were higher in bronchial (p = 0.02), but not bronchiolar, and in alveolar epithelial cells (both pneumocytes types I and II; p = 0.001) as well as in bronchial endothelial cells (p = 0.043) and neutrophils (p = 0.005) of subjects with COPD when compared with the levels in subjects without COPD (Figure 2) . The increased level of 4-HNE adducts in alveolar epithelium was inversely correlated with the FEV1 (r = -0.76, p < 0.05) if the results in all subjects were analyzed (Figure 3) . However, a trend toward an inverse relationship was found between the levels of 4-HNE in bronchial epithelium and FEV1 (r = -0.51; p = 0.075). Furthermore, the level of 4-HNE in bronchial endothelium and neutrophils correlated significantly with FEV1 (r = -0.61, p = 0.028; r = -0.56, p = 0.048, respectively), in an analysis of the data from all subjects.



View larger version (106K):
[in this window]
[in a new window]
  		Figure 1. Photographs from immunostaining for 4-HNE in lung tissue from subjects with and without COPD. (A) Non-COPD, bronchial; (B) Non-COPD, alveolar; (C) COPD, bronchial; (D) COPD, alveolar. Thin arrows point to pneumocyte types I and II. Thick arrows indicate alveolar macrophages. L = airway lumen; BV = blood vessel. Note the intensely stained neutrophils in the blood vessels in Figure 1C. Original magnification: x200.

 


View larger version (13K):
[in this window]
[in a new window]
  		Figure 2. Immunostaining scores for 4-HNE adducts per subject and cell type in bronchial (br) and alveolar (alv) tissue. (A) Epithelial cells and (B) endothelial cells and neutrophils. Open circles represent subjects without COPD (N), and closed circles represent subjects with COPD (O). The mean is indicated; significance levels (p values) for differences between the indicated groups are also shown.

 


View larger version (14K):
[in this window]
[in a new window]
  		Figure 3. Correlations between the levels of 4-HNE adducts in alveolar epithelium with FEV1 levels in subjects with (closed circles) and without (open circles) airway obstruction. Correlation (r) and significance level (p value) are indicated.

 
Recently, we showed in the same subject groups, on tissue sections adjacent to the sections used in this study, enhanced levels of TGF-ß1 and {gamma}-GCS mRNA in bronchiolar and alveolar epithelium in patients with COPD (20, 25). For the present study, historical data for TGF-ß1 and {gamma}-GCS expression from these studies were used. Upon correction for the number of patients included in the present study, these levels remained significantly enhanced in COPD for TGF-ß1 mRNA (1.6 times, p < 0.001) and protein (2.9 times, p = 0.005) as well as {gamma}-GCS mRNA (1.9 times, p = 0.024) (data not shown). Because TGF-ß1 was reported to increase ROS production (21, 22) and ROS has been shown to induce {gamma}-GCS-heavy subunit (HS) mRNA in alveolar epithelial cells (28), we examined the relationship of 4-HNE adduct levels with TGF-ß1 and {gamma}-GCS mRNA levels in airway and in alveolar epithelium. We found a significant correlation between the levels of 4-HNE adducts in bronchial and alveolar epithelium and TGFß1 mRNA (r = 0.62 and r = 0.51, respectively, p < 0.05) (Figures 4A and 4B) , and bronchial protein levels (r = 0.61, p = 0.015) (Figure 5) , as well as alveolar {gamma}-GCS mRNA (r = 0.63, p < 0.004) (Figure 6) in an analysis of data from all subjects. No significant correlation was found between 4-HNE and {gamma}-GCS mRNA in bronchial epithelium (r = 0.42, p = 0.16). Similarly, there was no significant correlation between 4-HNE adducts and TGFß1 protein levels in alveolar epithelium (r = 0.47, p = 0.1).




View larger version (27K):
[in this window]
[in a new window]
  		Figure 4. Correlations between the levels of 4-HNE adducts and TGF-ß1 mRNA expression both in bronchial (A) and alveolar (B) epithelium. Open circles represent data from subjects without COPD, and closed circles represent data from subjects with COPD. Correlation (r) and significance level (p value) are given.

 


View larger version (13K):
[in this window]
[in a new window]
  		Figure 5. Correlations between the levels of 4-HNE adducts and TGF-ß1 protein expression in bronchial epithelium. Open circles represent data from subjects without COPD, and closed circles represent data from subjects with COPD. Correlation (r) and significance level (p value) are given.

 


View larger version (12K):
[in this window]
[in a new window]
  		Figure 6. Correlations between the levels of 4-HNE adducts and {gamma}-GCS mRNA expression in alveolar epithelium. Open circles represent data from subjects without COPD, and closed circles represent data from subjects with COPD. Correlation (r) and significance level (p value) are given.

 

   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
The oxidant burden in the lungs is enhanced in smokers and in patients with COPD due to oxidants in cigarette smoke and by the release of ROS from airspace leukocytes (6, 7). A consequence of this increased burden may be lipid peroxidation in the lungs. 4-HNE is a highly reactive and specific diffusible end product of lipid peroxidation. In this study, we showed increased levels of 4-HNE adducts in lung epithelial and endothelial cells as well as neutrophils in subjects with COPD, compared with levels in subjects without COPD. We also found that the increased level of 4-HNE adducts in alveolar epithelium, airway endothelium, and neutrophils was inversely correlated with FEV1. We have previously shown high levels of lipid peroxidation products (thiobarbituric acid reactive substances) in blood and lung epithelial lining fluid of patients with COPD (29, 30). The levels of thiobarbituric acid reactive substances have also been shown to be elevated in breath condensate and lungs of smokers and patients with stable COPD (31–34). Further confirmation of oxidative stress and lipid peroxidation in patients with COPD comes from reports on elevated levels of 8-isoprostanes and hydrocarbons such as ethane and pentane in exhaled air condensate in healthy smokers and in patients with COPD (9–12). Isoprostanes (a member of F2-isoprostane) are stable end products of nonenzymatic lipid peroxidation of arachidonic acid (35). The increased levels of F2-isoprostane in exhaled breath condensate were found to be inversely correlated with airflow obstruction (9). Taken together, these data suggest a role for lipid peroxidation (specifically, the generation of 4-HNE adducts and F2-isoprostane) in airway obstruction in COPD.

The results from the present study show that increased levels of 4-HNE adducts are associated with increased levels of both TGF-ß1 protein and mRNA. We, and others, have shown that TGF-ß1 is localized mainly in bronchiolar and alveolar epithelium and macrophages, and that the epithelial TGF-ß1 expression is higher in subjects with COPD, compared with those without COPD (20, 36). TGF-ß1 is a multifunctional growth factor that modulates cellular proliferation, differentiation, and tissue repair (19). TGF-ß1 is also a chemoattractant and mitogen for fibroblasts and fibroblast-like cells, and it stimulates the synthesis and deposition of extracellular matrix. TGF-ß1 has been suggested to increase oxidative stress, leading to the generation of lipid peroxidation products, on the basis of the observation that it increases the cellular release of hydrogen peroxide from endothelial cells (21, 22). Vice versa, lipid peroxidation products may also affect the expression of TGF-ß1, as has been shown in experiments with 4-HNE (37). 4-HNE has been shown to induce cellular stress responses such as cell signaling via the mitogen-activated protein kinase pathways leading to the induction of AP-1–mediated genes (16, 17). In vitro studies showed that 4-HNE increased TGF-ß1 expression by a mechanism dependent on the activation of AP-1 in macrophages (37). Hence, it is likely that lipid peroxidation induces TGF-ß1 expression in lungs of patients with COPD. However, there are other confounding factors such as differences in local tumor necrosis factor-{alpha} levels, which have been shown to be higher in patients with COPD (38), that may be associated with increased oxidative stress (formation of 4-HNE) and {gamma}-GCS expression (25, 28, 39).

In turn, TGF-ß1 also causes a marked decrease in glutathione levels in endothelial and alveolar epithelial cells and downregulates {gamma}-GCS mRNA levels in vitro in alveolar epithelial cells (23). 4-HNE has also been shown to induce {gamma}-GCS mRNA in alveolar epithelial cells (24). We also showed increased {gamma}-GCS expression in lungs of patients with COPD (25). 4-HNE–mediated induction of {gamma}-GCS gene was associated with the mitogen-activated protein kinase signaling pathways (24). It is also known that cigarette smoke induces {gamma}-GCS gene expression via the activation of AP-1 in alveolar epithelial cells (40). In this study, we found a significant correlation between 4-HNE adduct levels and {gamma}-GCS mRNA in airway or alveolar epithelium in subjects without or with COPD. The induction of {gamma}-GCS may be an important adaptive response of the alveolar epithelium to oxidative stress. This suggests that 4-HNE is a second messenger that may play a role in the regulation of expression of the protective {gamma}-GCS gene and also a variety of other genes like TGF-ß1, cyclooxygenase 2, and monocyte chemoattractant protein-1 that were reported to be implicated in the pathogenesis of COPD (41, 42) (Figure 7) . An imbalance of an array of redox-regulated antioxidant versus proinflammatory genes might be associated with the susceptibility or tolerance to disease (34).



View larger version (22K):
[in this window]
[in a new window]
  		Figure 7. Proposed model of the mechanism of cigarette smoke–mediated oxidative stress (formation of 4-HNE) in lung inflammation.

 
In conclusion, this study showed that in smokers with and without COPD, 4-HNE is formed in airway epithelial cells, endothelial cells, as well as neutrophils and macrophages. Higher 4-HNE adduct levels in epithelial cells were found in subjects with COPD, compared with levels in subjects without COPD. Thus, elevated levels of 4-HNE may be the end product of oxidant stress imposed by cigarette smoking, which appears to be more pronounced in those who developed COPD. Furthermore, we found a significant correlation of 4-HNE adduct levels with TGF-ß1 protein and mRNA, {gamma}-GCS mRNA in airway or alveolar epithelium, and FEV1 in subjects without or with COPD. This indicates that the generation of 4-HNE, TGF-ß1 mRNA, and {gamma}-GCS may be associated with the imbalance of proinflammatory and protective antioxidant responses that occurs in lungs of patients with COPD. In turn, this points to a potential role for 4-HNE in the signaling events involved in lung inflammation leading to the development of COPD.


   FOOTNOTES
 
This work was supported by Aventis Pharmaceuticals, UK, the Cunningham Trust, UK, the Netherlands Asthma Foundation (Grant 98.12), and the European Union–funded COPD GENE SCAN project no. QRLT-2000-01012.

Received in original form October 29, 2001; accepted in final form May 7, 2002


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. American Thoracic Society. Standards for the diagnosis and care of patients with chronic obstructive pulmonary disease. Am J Respir Crit Care Med 1995;152:S77–S120.
  2. British Thoracic Society. Guidelines for the management of chronic obstructive pulmonary disease. Thorax 1997;52(Suppl 5):S1–S28.[Medline]
  3. Saetta M. Airway inflammation in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 1999;160:S17–S20.[Abstract/Free Full Text]
  4. Rahman I, MacNee W. Role of oxidants/antioxidants in smoking-induced airways diseases. Free Radic Biol Med 1996;21:669–681.[CrossRef][Medline]
  5. Pryor WA, Stone K. Oxidants in cigarette smoke: radicals, hydrogen peroxides, peroxynitrate, and peroxynitrite. Ann NY Acad Sci 1993; 686:12–28.[Medline]
  6. Rahman I, MacNee W. Lung glutathione and oxidative stress: implications in cigarette smoke-induced airway disease. Am J Physiol 1999; 277:L1067–L1088.[Abstract/Free Full Text]
  7. Repine JE, Bast A, Lankhorst I, and the Oxidative Stress Study Group. Oxidative stress in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 1997;156:341–357.[Free Full Text]
  8. Cross CE, van der Vliet A, Eiserich JP. Cigarette smokers and oxidant stress: a continuing mystery. Am J Clin Nutr 1998;67:184–185.[Medline]
  9. Montuschi P, Collins JV, Ciabattoni G, Lazzeri N, Corradi M, Kharitonov SA, Barnes PJ. Exhaled 8-isoprostane as an in vivo biomarker of lung oxidative stress in patients with COPD and healthy smokers. Am J Respir Crit Care Med 2000;162:1175–1177.[Abstract/Free Full Text]
  10. Paredi P, Kharitonov SA, Leak D, Ward S, Cramer D, Barnes PJ. Exhaled ethane, a marker of lipid peroxidation, is elevated in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 2000;162:369–373.[Abstract/Free Full Text]
  11. Habib MP, Clements NC, Garewal HS. Cigarette smoking and ethane exhalation in humans. Am J Respir Crit Care Med 1995;151:1368–1372.[Abstract]
  12. Euler DE, Dave SJ, Guo H. Effect of cigarette smoking on pentane excretion in alveolar breath. Clin Chem 1996;42:303–308.[Abstract/Free Full Text]
  13. Gutteridge JML. Lipid peroxidation and antioxidants as biomarkers of tissue damage. Clin Chem 1995;41:1819–1828.[Abstract/Free Full Text]
  14. Esterbauer H, Schaur RJ, Zollner H. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med 1991;11:81–128.[CrossRef][Medline]
  15. Uchida K, Szweda LI, Chae HZ, Stadtman ER. Immunochemical detection of 4-hydroxynonenal protein adducts in oxidized hepatocytes. Proc Natl Acad Sci USA 1993;90:8742–8746.[Abstract/Free Full Text]
  16. Uchida K, Shiraishi M, Naito Y, Torii Y, Nakamura Y, Osawa T. Activation of stress signaling pathways by the end product of lipid peroxiation. J Biol Chem 1999;274:2234–2242.[Abstract/Free Full Text]
  17. Leonarduzzi G, Arkan MC, Basaga H, Chiarpotto E, Sevanian A, Poli G. Lipid oxidation products in cell signaling. Free Radic Biol Med 2000;28:1760–1768.
  18. Parola M, Bellomo G, Robino G, Barrera G, Dianzani MU. 4-Hydroxynonenal as a biological signal: molecular basis and pathophysiological implications. Antioxid Redox Signal 1999;1:255–284.[Medline]
  19. Border WA, Ruoslahti E. Transforming growth factor-ß1 in disease: the dark side of tissue repair. J Clin Invest 1992;90:1–7.
  20. de Boer WI, van Schadewijk A, Sont JK, Sharma HS, Stolk J, Hiemstra PS, van Krieken JHJM. Transforming growth factor-ß1 and recruitment of macrophages and mast cells in airways in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 1998;158:1951–1957.[Abstract/Free Full Text]
  21. Das SK, Fanburg BL. TGF-ß1 produces a ‘prooxidant’ effect on bovine pulmonary artery endothelial cells in culture. Am J Physiol 1991; 262:L249–L254.
  22. White AC, Das SK, Fanburg BL. Reduction of glutathione is associated with growth restriction and enlargement of bovine pulmonary artery endothelial cells produced by transforming growth factor-ß1. Am J Respir Cell Mol Biol 1992;6:364–368.
  23. Arsalane K, Dubois CM, Muanza T, Begin R, Boudreau F, Asselin C, Cantin AM. Transforming growth factor-ß1 is a potent inhibitor of glutathione synthesis in the lung epithelial cell line A549: transcriptional effect on the GSH rate-limiting enzyme {gamma}-glutamylcysteine synthetase. Am J Respir Cell Mol Biol 1997;17:599–697.[Abstract/Free Full Text]
  24. Liu RM, Borok Z, Forman HJ. 4-Hydroxy-2-nonenal increases {gamma}-glutamylcysteine synthetase gene expression in alveolar epithelial cells. Am J Respir Cell Mol Biol 2001;24:499–505.[Abstract/Free Full Text]
  25. Rahman I, Van Schadewijk AAM, Hiemstra PS, Stolk J, Van Krieken JHJM, MacNee W, De Boer WI. Localisation of {gamma}-glutamylcysteine synthetase messenger RNA expression in lungs of smokers and patients with chronic obstructive pulmonary disease. Free Radic Biol Med 2000;28:920–925.[CrossRef][Medline]
  26. De Boer WI, Sont JK, van Schadewijk A, Stolk J, van Krieken JH, Hiemstra PS. Monocyte chemoattractant protein 1, interleukin 8, and chronic airways inflammation in COPD. J Pathol 2000;190:619–626.[CrossRef][Medline]
  27. Toyokuni S. Reactive oxygen species-induced molecular damage and its application in pathology. Pathol Int 1999;49:91–102.[CrossRef][Medline]
  28. Rahman I, MacNee W. Oxidative stress and regulation of glutathione synthesis in lung inflammation. Eur Respir J 2000;16:534–554.[Abstract]
  29. Rahman I, Morrison D, Donaldson K, MacNee W. Systemic oxidative stress in asthma, COPD, and smokers. Am J Respir Crit Care Med 1996;154:1055–1060.[Abstract]
  30. Morrison D, Rahman I, Lannan S, MacNee W. Epithelial permeability, inflammation, and oxidant stress in the airspaces of smokers. Am J Respir Crit Care Med 1999;159:473–479.[Abstract/Free Full Text]
  31. Lapenna D, Gioia SD, Mezzetti A, Ciofani G, Consoli A, Marzio L, Cuccurullo F. Cigarette smoke, ferritin, and lipid peroxidation. Am J Respir Crit Care Med 1995;151:431–435.[Abstract]
  32. Nowak D, Kasielski M, Antczak A, Pietras T, Bialasiewicz P. Increased content of thiobarbiturate reactive acid substances in hydrogen peroxide in the expired breath condensate of patients with stable chronic obstructive pulmonary disease: no significant effect of cigarette smoking. Respir Med 1999;93:389–396.[CrossRef][Medline]
  33. Fahn H, Wang L, Kao S, Chang S, Huang M, Wei Y. Smoking-associated mitochondrial DNA mutation and lipid peroxidation in human lung tissue. Am J Respir Cell Mol Biol 1998;19:901–909.[Abstract/Free Full Text]
  34. MacNee W, Rahman I. Oxidants and antioxidant in COPD: therapeutic targets. Am J Respir Crit Care Med 1999;160:S1–S8.[Free Full Text]
  35. Lawson JA, Rokach J, FitzGerald GA. Isoprostanes: formation, analysis and use as indices of lipid peroxidation in vivo. J Biol Chem 1999; 274:24441–24444.[Free Full Text]
  36. Takizawa H, Tanaka M, Takami K, Ohtoshi T, Ito K, Satoh M, Okada Y, Yamasawa F, Nakahara K, Umeda A. Increased expression of transforming growth factor-ß1 in small airway epithelium from tobacco smokers and patients with chronic obstructive pulmonary disease (COPD). Am J Respir Crit Care Med 2001;163:1476–1483.[Abstract/Free Full Text]
  37. Leonarduzzi G, Scavazza A, Biasi F, Chiarpotto E, Camandola S, Vogl S, Dargel R, Poli G. The lipid peroxidation end product 4-hydroxy-2,3-nonenal up-regulates transforming growth factor-ß1 expression in the macrophage lineage: a link between oxidative injury and fibrosclerosis. FASEB J 1997;11:851–857.[Abstract]
  38. Keatings VM, Collins PD, Scott DN, Barnes PJ. Differences in interleukin-8 and tumor necrosis factor-alpha in induced sputum from patients with chronic obstructive pulmonary disease or asthma. Am J Respir Crit Care Med 1992;153:530–534.[Abstract]
  39. Rahman I, Antonicelli F, MacNee W. Molecular mechanism of the regulation of glutathione synthesis by tumour necrosis factor-{alpha} and dexamethasone in human alveolar epithelial cells. J Biol Chem 1999;274: 5088–5096.[Abstract/Free Full Text]
  40. Rahman I, Smith CAD, Lawson MF, Harrison DJ, MacNee W. Induction of {gamma}-glutamylcysteine synthetase by cigarette smoke is associated with AP-1 in human alveolar epithelial cells. FEBS Lett 1996;396:21–25.[CrossRef][Medline]
  41. Bellocq A, Azoulay E, Marullo S, Flahault A, Fouqueray B, Philippe C, Cadranel J, Baud L. Reactive oxygen and nitrogen intermediates increase transforming growth factor-ß1 release from human epithelial alveolar cells through two different mechanisms. Am J Respir Cell Mol Biol 1999;21:128–136.[Abstract/Free Full Text]
  42. Kumagai T, Kawamoto Y, Nakamura Y, Hatayama I, Satoh K, Osawa T, Uchida K. 4-hydroxy-2-nonenal, the end product of lipid peroxidation, is a specific inducer of cyclooxygenase-2 gene expression. Biochem Biophys Res Commun 2000;273:437–441.[CrossRef][Medline]



This article has been cited by other articles:


Home page
 BloodHome page
P. Margutti, P. Matarrese, F. Conti, T. Colasanti, F. Delunardo, A. Capozzi, T. Garofalo, E. Profumo, R. Rigano, A. Siracusano, et al.
Autoantibodies to the C-terminal subunit of RLIP76 induce oxidative stress and endothelial cell apoptosis in immune-mediated vascular diseases and atherosclerosis
Blood, May 1, 2008; 111(9): 4559 - 4570.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
S. Rajendrasozhan, S.-R. Yang, V. L. Kinnula, and I. Rahman
SIRT1, an Antiinflammatory and Antiaging Protein, Is Decreased in Lungs of Patients with Chronic Obstructive Pulmonary Disease
Am. J. Respir. Crit. Care Med., April 15, 2008; 177(8): 861 - 870.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. Trevisani, J. Siemens, S. Materazzi, D. M. Bautista, R. Nassini, B. Campi, N. Imamachi, E. Andre, R. Patacchini, G. S. Cottrell, et al.
4-Hydroxynonenal, an endogenous aldehyde, causes pain and neurogenic inflammation through activation of the irritant receptor TRPA1
PNAS, August 14, 2007; 104(33): 13519 - 13524.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. Rev.Home page
T. Yoshida and R. M. Tuder
Pathobiology of Cigarette Smoke-Induced Chronic Obstructive Pulmonary Disease
Physiol Rev, July 1, 2007; 87(3): 1047 - 1082.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
J.-i. Machiya, Y. Shibata, K. Yamauchi, N. Hirama, T. Wada, S. Inoue, S. Abe, N. Takabatake, M. Sata, and I. Kubota
Enhanced Expression of MafB Inhibits Macrophage Apoptosis Induced by Cigarette Smoke Exposure
Am. J. Respir. Cell Mol. Biol., April 1, 2007; 36(4): 418 - 426.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol PatholHome page
J.L. Wright and A. Churg
Current Concepts in Mechanisms of Emphysema
Toxicol Pathol, January 1, 2007; 35(1): 111 - 115.
[Abstract] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
A. Singh, T. Rangasamy, R. K. Thimmulappa, H. Lee, W. O. Osburn, R. Brigelius-Flohe, T. W. Kensler, M. Yamamoto, and S. Biswal
Glutathione Peroxidase 2, the Major Cigarette Smoke-Inducible Isoform of GPX in Lungs, Is Regulated by Nrf2
Am. J. Respir. Cell Mol. Biol., December 1, 2006; 35(6): 639 - 650.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. V. Usatyuk, N. L. Parinandi, and V. Natarajan
Redox Regulation of 4-Hydroxy-2-nonenal-mediated Endothelial Barrier Dysfunction by Focal Adhesion, Adherens, and Tight Junction Proteins
J. Biol. Chem., November 17, 2006; 281(46): 35554 - 35566.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
R. M. Tuder
Aging and Cigarette Smoke: Fueling the Fire
Am. J. Respir. Crit. Care Med., September 1, 2006; 174(5): 490 - 491.
[Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
T. Sato, K. Seyama, Y. Sato, H. Mori, S. Souma, T. Akiyoshi, Y. Kodama, T. Mori, S. Goto, K. Takahashi, et al.
Senescence Marker Protein-30 Protects Mice Lungs from Oxidative Stress, Aging, and Smoking
Am. J. Respir. Crit. Care Med., September 1, 2006; 174(5): 530 - 537.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
M.-C. L. Machado, J. A. Krishnan, S. A. Buist, A. L. Bilderback, G. P. Fazolo, M. G. Santarosa, F. Queiroga Jr., and W. M. Vollmer
Sex Differences in Survival of Oxygen-dependent Patients with Chronic Obstructive Pulmonary Disease
Am. J. Respir. Crit. Care Med., September 1, 2006; 174(5): 524 - 529.
[Abstract] [Full Text] [PDF]

Home page
 Proc Am Thorac SocHome page
R. M. Tuder, T. Yoshida, W. Arap, R. Pasqualini, and I. Petrache
State of the Art. Cellular and Molecular Mechanisms of Alveolar Destruction in Emphysema: An Evolutionary Perspective
Proceedings of the ATS, August 1, 2006; 3(6): 503 - 510.
[Abstract] [Full Text] [PDF]

Home page
 Eur Respir JHome page
I. Rahman and I. M. Adcock
Oxidative stress and redox regulation of lung inflammation in COPD.
Eur. Respir. J., July 1, 2006; 28(1): 219 - 242.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
C. J. Baglole, S. M. Bushinsky, T. M. Garcia, A. Kode, I. Rahman, P. J. Sime, and R. P. Phipps
Differential induction of apoptosis by cigarette smoke extract in primary human lung fibroblast strains: implications for emphysema
Am J Physiol Lung Cell Mol Physiol, July 1, 2006; 291(1): L19 - L29.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
S.-R. Yang, A. S. Chida, M. R. Bauter, N. Shafiq, K. Seweryniak, S. B. Maggirwar, I. Kilty, and I. Rahman
Cigarette smoke induces proinflammatory cytokine release by activation of NF-{kappa}B and posttranslational modifications of histone deacetylase in macrophages
Am J Physiol Lung Cell Mol Physiol, July 1, 2006; 291(1): L46 - L57.
[Abstract] [Full Text] [PDF]

Home page
 ThoraxHome page
K Nagai, T Betsuyaku, T Kondo, Y Nasuhara, and M Nishimura
Long term smoking with age builds up excessive oxidative stress in bronchoalveolar lavage fluid
Thorax, June 1, 2006; 61(6): 496 - 502.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
I. Dalle-Donne, R. Rossi, R. Colombo, D. Giustarini, and A. Milzani
Biomarkers of Oxidative Damage in Human Disease
Clin. Chem., April 1, 2006; 52(4): 601 - 623.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
R. F. Foronjy, O. Mirochnitchenko, O. Propokenko, V. Lemaitre, Y. Jia, M. Inouye, Y. Okada, and J. M. D'Armiento
Superoxide Dismutase Expression Attenuates Cigarette Smoke- or Elastase-generated Emphysema in Mice
Am. J. Respir. Crit. Care Med., March 15, 2006; 173(6): 623 - 631.
[Abstract] [Full Text] [PDF]

Home page
 Am J EpidemiolHome page
H. M. Ochs-Balcom, B. J. B. Grant, P. Muti, C. T. Sempos, J. L. Freudenheim, R. W. Browne, M. Trevisan, L. Iacoviello, P. A. Cassano, and H. J. Schunemann
Oxidative Stress and Pulmonary Function in the General Population
Am. J. Epidemiol., December 15, 2005; 162(12): 1137 - 1145.
[Abstract] [Full Text] [PDF]

Home page
 Proc Am Thorac SocHome page
W. MacNee
Pathogenesis of Chronic Obstructive Pulmonary Disease
Proceedings of the ATS, November 1, 2005; 2(4): 258 - 266.
[Abstract] [Full Text] [PDF]

Home page
 Proc Am Thorac SocHome page
I. M. Adcock and K. Ito
Glucocorticoid Pathways in Chronic Obstructive Pulmonary Disease Therapy
Proceedings of the ATS, November 1, 2005; 2(4): 313 - 319.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
R. D. Wang, J. L. Wright, and A. Churg
Transforming Growth Factor-{beta}1 Drives Airway Remodeling in Cigarette Smoke-Exposed Tracheal Explants
Am. J. Respir. Cell Mol. Biol., October 1, 2005; 33(4): 387 - 393.
[Abstract] [Full Text] [PDF]

Home page
 ERRHome page
W. MacNee
Treatment of stable COPD: antioxidants
Eur. Respir. Rev., September 1, 2005; 14(94): 12 - 22.
[Abstract] [Full Text] [PDF]

Home page
 Eur Respir JHome page
P. J. Barnes and R. A. Stockley
COPD: current therapeutic interventions and future approaches
Eur. Respir. J., June 1, 2005; 25(6): 1084 - 1106.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Crit. Care Med.Home page
H. Yasuda, M. Yamaya, K. Nakayama, S. Ebihara, T. Sasaki, S. Okinaga, D. Inoue, M. Asada, M. Nemoto, and H. Sasaki
Increased Arterial Carboxyhemoglobin Concentrations in Chronic Obstructive Pulmonary Disease
Am. J. Respir. Crit. Care Med., June 1, 2005; 171(11): 1246 - 1251.
[Abstract] [Full Text] [PDF]

Home page
 Proc Am Thorac SocHome page
N. Voelkel and L. Taraseviciene-Stewart
Emphysema: An Autoimmune Vascular Disease?
Proceedings of the ATS, April 1, 2005; 2(1): 23 - 25.
[Abstract] [Full Text] [PDF]

Home page
 Proc Am Thorac SocHome page
W. MacNee
Pulmonary and Systemic Oxidant/Antioxidant Imbalance in Chronic Obstructive Pulmonary Disease
Proceedings of the ATS, April 1, 2005; 2(1): 50 - 60.
[Abstract] [Full Text] [PDF]

Home page
 Eur Respir JHome page
P. J. Barnes, I. M. Adcock, and K. Ito
Histone acetylation and deacetylation: importance in inflammatory lung diseases
Eur. Respir. J., March 1, 2005; 25(3): 552 - 563.
[Abstract]