This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Koyama, S. Articles by Izumi, T. Search for Related Content PubMed PubMed Citation Articles by Koyama, S. Articles by Izumi, T.

Decreased Level of Vascular Endothelial Growth Factor in Bronchoalveolar Lavage Fluid of Normal Smokers and Patients with Pulmonary Fibrosis

National Chuushin Matsumoto Hospital, Matsumoto; JA Shinnmachi Hospital, Shinnmachi; The Second Department of Surgery, Shinshu University School of Medicine, Matsumoto; and Department of Respiratory Medicine, Graduate School of Medicine, Kyoto University, Kyoto, Japan

Correspondence and requests for reprints should be addressed to Sekiya Koyama, M.D., Pulmonary Section, National Chuushin Matsumoto Hospital, 811 Kotobuki Toyooka, Matsumoto 399-0021, Japan. E-mail: koyama{at}ka3.so-net.ne.jp

	ABSTRACT

TOP ABSTRACT INTRODUCTION METHODS RESULTS DISCUSSION REFERENCES

 
Vascular endothelial growth factor (VEGF) plays multifunctional roles in both the development of vasculature and the maintenance of vascular function. A decrease in VEGF reduces angiogenesis and induces apoptosis of vascular endothelial cells. Inhibition of the VEGF receptor causes endothelial cell apoptosis and emphysema. We postulated that VEGF concentrations might be reduced in patients with chronic lung diseases. The level of VEGF was evaluated by enzyme-liked immunosorbant assay in bronchoalveolar lavage fluid (BALF) from normal smokers, nonsmoking volunteers, idiopathic pulmonary fibrosis, pulmonary fibrosis associated with a connective tissue disease, and sarcoidosis. The isoforms of VEGF in BALF were determined by high-performance liquid chromatography. VEGF in nonsmoking volunteers was detectable at a high concentration. In contrast, VEGF in most of the normal smokers was below the detectable limit. The VEGF found in nonsmoking volunteers BALF was VEGF165. VEGF was significantly decreased in idiopathic pulmonary fibrosis, pulmonary fibrosis associated with a connective tissue disease, and sarcoidosis compared with nonsmoking volunteers. The smoking patients showed a further decrease in VEGF. These data suggest that the decrease in VEGF in smokers and patients with chronic lung diseases may reduce angiogenesis and induce apoptosis of vascular endothelial cells.

Key Words: vascular endothelial growth factor • bronchoalveolar lavage • smoking

	INTRODUCTION

TOP ABSTRACT INTRODUCTION METHODS RESULTS DISCUSSION REFERENCES

 
Vascular endothelial growth factor (VEGF) is a dimorphic glycoprotein with a molecular weight of 34,000 to 42,000, consisting of two disulfide-linked peptide chains having identical N termini (1, 2). VEGF possesses potent vascular permeability-enhancing activity. On a molar basis, it is 50,000 times more potent than histamine (1). VEGF was first identified in tumor cell-conditioned media (3) and has been purified to homogeneity from guinea pig and human sources (1, 4). VEGF acts directly on cultured vascular endothelium to induce a rapid increase in free cytosole calcium, apparently by activating a phosphoinositide-specific phospholipase C (5). It stimulates the release of von Willebrand factor from endothelial cells (5) and induces the expression of endothelial cell tissue factor activity (6). Concurrently with these studies, a protein selectively mitogenic for vascular endothelial cells was purified from cell culture, and this protein turned out to be the same substance that induced vascular permeability-enhancing activity (7, 8). VEGF is chemotactic for endothelial cells (9) and enhances collagenase (10) and urokinase receptor expression in endothelial cells (11). The existence of multiple activities embodied in the same protein suggests that VEGF may play multifunctional roles in both the development of vasculature and the maintenance of vascular structure and function.

The gene for human VEGF is organized into eight exons. As a result of alternative splicing, at least four transcripts have been detected, encoding mature monomeric VEGF containing 121, 165, 189, and 206 amino acid residues (VEGF121, VEGF165, VEGF189, and VEGF206), each preceded by a 26-amino acid signal peptide (12). VEGF121 and VEGF165 are diffusible proteins that are secreted into medium. VEGF189 and VEGF206 have high affinity for heparin and are mostly bound to heparin-containing proteoglycans in the extracellular matrix (13). Transfection studies demonstrate that VEGF121 and VEGF165 are secreted and have both mitogenic and peameability-inducing activities. In contrast, VEGF189 and VEGF206 remain primarily cell associated and lack the mitogenic activity of the smaller form (14, 15), suggesting that VEGF165 and VEGF121 are pivotal forms of VEGF. Among several peptide growth factors implicated in angiogenesis, including transforming growth factor-ß, platelet-derived growth factor, and VEGF, only VEGF is specifically mitogenic for endothelial cells, having no mitogenic activity for other cell types (16). VEGF mRNA has been found in embryonic tissues, including lung, kidney, and placenta, during organ angiogenesis (14, 17). Although VEGF transcripts are located in epithelial cells, smooth muscle cells, and macrophages in various tissues, in normal adult lung, expression has been mainly in alveolar epithelial cells (18).

The mechanisms that maintain lung alveolar capillary structure and function are not completely known. Recently, Kasahara and coworkers reported that inhibition of VEGF receptors causes endothelial cell apoptosis and emphysema, suggesting that VEGF receptor signaling is required for maintenance of the alveolar structure and that alveolar septal cell apoptosis contributes to the pathogenesis of emphysema (19). If VEGF is necessary for the maintenance of healthy pulmonary capillary endothelium and blocking its receptor causes emphysema, then a reduced level of VEGF in the epithelial lining fluid from smoking may be a mechanism whereby cigarette smoking causes emphysema by inducing endothelial apoptosis and alveolar wall destruction. Thus, in this study, we evaluated whether smoking and other chronic lung diseases are associated with a reduction in the concentration of VEGF in bronchoalveolar lavage fluid (BALF).

	METHODS

TOP ABSTRACT INTRODUCTION METHODS RESULTS DISCUSSION REFERENCES

 
Normal Control Subjects
Analysis of VEGF and cellular constituents of BALF was performed in 27 healthy volunteers and in patients with idiopathic pulmonary fibrosis (IPF), pulmonary fibrosis associated with a connective tissue disease (PF-CTD), and sarcoidosis (Sar) (Table 1) . Sixteen were never-smokers, and 11 were current smokers. Smoking history varied from 5.5 to 13.4 years (8.9 ± 3.8 years, average). They smoked 1.2 ± 0.3 packs per day. These subjects had no respiratory symptoms, normal chest radiographs, and normal spirometric test results, lung volume, and diffusing capacity for carbon monoxide. None of the subjects had evidence of hypersensitivity pneumonia, left ventricular dysfunction, cardiac valvular diseases, or significant occupational exposures. Informed consent was obtained from each patient and healthy volunteer, and the protocol was approved by the Institutional Human Subjects Review Committee.

Patients with IPF
Fourteen patients with IPF were evaluated (Table 1). IPF was diagnosed on the basis of a comparable history, physical examination, chest radiograph, and pulmonary physiologic evaluation. A confirmatory open-lung biopsy was performed in all 14 subjects. Thirteen out of 14 patients had usual interstitial pneumonia (UIP), and one of the patients had nonspecific interstitial pneumonia. Two of the patients with IPF were receiving treatment at the time of lavage (prednisolone alone). Eleven patients were either never-smokers or ex-smokers (quit at least 6 months before), and there were three current smokers.

Patients with PF-CTD
Twenty-one patients with PF-CTD were examined. Seven patients had progressive systemic sclerosis (seven, UIP). Six had dermatomyositis/polymyositis (four, UIP; two, nonspecific interstitial pneumonia). One had mixed connective tissue disease (one, UIP). Two had rheumatoid arthritis (one, UIP; one, bronchiolitis obliterance organizing pneumonia), and five had Sjogren's syndrome (four, UIP; one, bronchiolitis obliterance organizing pneumonia). Ten patients with PF-CTD were receiving treatment at the time of lavage (prednisolone alone). Sixteen patients were never-smokers or were ex-smokers (quit at least 6 months before), and five patients were current smokers.

Patients with Sar
Fourteen patients with Sar participated in this study. These patients were diagnosed on the basis of a comparable history, physical examination, chest radiograph, typical BALF lymphocyte surface markers, and pulmonary physiologic evaluation. A confirmatory open-lung biopsy was performed in 14 subjects. Two patients were treated with oral steroids. Nine patients were never-smokers or ex-smokers (quit at least 6 months before), and five patients were current smokers.

Bronchoalveolar Lavage
Bronchoalveolar lavage was performed in an outpatient setting using previously reported methods (20). Differential counts on 400 cells were performed on Wright-Giemsa–stained preparations made from the pooled lavage fluids before centrifugation. The pooled lavage fluid was centrifuged at 400 x g (2,000 rpm) for 10 minutes to separate the cells from the supernatant. The supernatant was stored at -80°C until assayed.

Measurement of VEGF Protein in BALF
The concentrations of VEGF and albumin in the unconcentrated BALF were assayed by an enzyme-linked immunosorbant assay kit (Amersham, Little Chalfont, UK) and the bromocresol green method (Seiken, Tokyo, Japan), respectively. The minimum concentration of VEGF detected by this method was 15.6 pg/ml. Then we calculated VEGF concentration divided by albumin concentration (pg/mg) in BALF.

Determination of the Isoform of VEGF in BALF by High-Performance Liquid Chromatography
To determine the isoforms of VEGF in BALF, we used high-performance liquid chromatography (Waters 2690 separations Module; Waters Corp., Milford, MA) by using a TSK gel-G3000SW column (Tosoh Corporation, Tokyo, Japan) to separate each isoform of VEGF containing 121, 165, 189, and 206 amino acids by molecular weight difference. The supernatant was filtered by Samprep LH4 (0.5 µm; Millipore, Bedford, MA) before high-performance liquid chromatography separation. The sample was injected into the TSK gel-G3000SW column, and the column was eluted with 0.1 M phosphate buffer with 0.2 M sodium chloride, pH 7.2, at a flow rate of 0.5 ml/minute. The VEGF protein concentration of every other fraction sample was measured by enzyme-linked immunosorbant assay. Chymotripsinogen (25 kD; Serva Electrophoresis, Heidelberg, Germany) and egg albumin (45 kD; Serva Electrophoresis) were used as molecular markers. The human recombinant VEGF165 was also eluted under the same conditions, and the VEGF protein concentration of each fraction was measured to assess the fraction profile of VEGF165.

Statistics
The differences between groups were tested for significance using the Kruskal-Wallis test for VEGF comparison. The differences were tested by the values above the detectable limit of VEGF. In all cases, a p value of less than 0.05 was considered significant. Data in figures and tables are expressed as means ± SEM.

	RESULTS

TOP ABSTRACT INTRODUCTION METHODS RESULTS DISCUSSION REFERENCES

 
BALF Findings in Nonsmoking Volunteers and Normal Smokers
The total cell count in BALF was increased in normal smokers, IPF, PF-CTD, and Sar (Table 2) . The percentage of neutrophils was not increased in normal smokers compared with nonsmoking volunteers (Table 2). However, the percentage of neutrophils was increased in IPF and PF-CTD (Table 2). In IPF, PF-CTD, and Sar, there was a decrease in the percentage of macrophages and an increase in lymphocytes (Table 2).

View larger version (18K): [in this window] [in a new window]   Figure 1. The concentration of VEGF in the BALF of normal volunteers and patients with IPF (n = 14), PF-CTD (n = 21), and Sar (n = 14). Open circles represent nonsmokers. Open squares represent current smokers. Values are means ± SEM; p values were determined using the data from nonsmokers. Values under the detectable range were estimated at 156 pg/mg albumin for comparison.

	DISCUSSION

TOP ABSTRACT INTRODUCTION METHODS RESULTS DISCUSSION REFERENCES

 
It is reported that the level of VEGF in the epithelial lining fluid was even higher than that in the plasma of healthy volunteers (21). Our results also demonstrate that VEGF protein was present in epithelial lining fluid at high levels in healthy subjects. As determined by Northern blotting on a per actin basis, the order of relative abundance of VEGF mRNA in adult tissue is lung, kidney, liver, brain, and spleen in human, guinea pig, mouse, and rat tissues (22–24), suggesting that the function of VEGF in lung tissue may be critical to normal physiology and, thus, expression is highly conserved. In humans, guinea pigs, mice, and rats (22–24), cellular localization in the lung was observed primarily in the alveolar epithelial cells. Thus, these data support the concept that VEGF is highly compartmentalized in the lung as previously reported (21).

A more complete understanding of the functions of VEGF in the lung depends on several factors: the size and bioactivity of VEGF expressed, the presence of VEGF receptors on adjacent endothelial cells, and the knowledge of mechanisms of both mitogenic and permeability responses. Among them, the size of VEGF secreted is of importance. Although direct analysis of which VEGF forms are expressed has not been systematically performed in the all of the tissues and species in which VEGF has been identified, in human tissues, VEGF165 seems to be the predominant form, followed by VEGF121 and VEGF189 (25, 26). In this study, the isoform of VEGF was VEGF165, and other forms of VEGF were not detected in BALF. Because VEGF165 shows bioactivity and diffusibility to the receptors of endothelial cells (14, 15), VEGF165 in the epithelial lining fluid may regulate many endothelial cell functions.

VEGF is thought to be involved in the regulation of capillary function, mitogenesis, and permeability-enhancing effects of the lung. However, the finding that expression of VEGF mRNA occurs diffusely in the brain and is concentrated in the cerebellar granule cell layer suggests that neither the angiogenesis nor the permeability-inducing activity of VEGF is of primary importance in the brain. VEGF has been reported to stimulate hexose transport in isolated peripheral endothelial cells (27). VEGF may regulate the transport of small molecules (e.g., glucose and amino acids) to support the increased requirement for energy of epithelial cells. The fact that the decreased level of VEGF not only reduces endothelial cell growth but also induces endothelial cell apoptosis (28) may support mechanisms underlying the decrease in the lung vascular volume observed in pulmonary emphysema. The decreased level of VEGF in normal smokers suggests that the smokers' lungs may have impaired capillary function, mitogenesis, and energy support.

The present authors previously reported that airway epithelial cells express and secrete VEGF165 in response to a variety of stimuli, including interleukin-1ß, tumor necrosis factor-, smoke extract, and neutrophil elastase (29). Thus, it is expected that normal smokers and patients with pulmonary fibrosis would have a higher concentration of VEGF in BALF. However, the lower levels of VEGF in pulmonary fibrosis were consistent with the previous report (30). Although the mechanisms of the decreased levels of VEGF in BALF are uncertain, the epithelial cell apoptosis and cellular injury observed in normal smokers and patients with pulmonary fibrosis (31–33) may be one of the underlying mechanisms that explains the decrease in the release of VEGF. To support this hypothesis, Klekamp and colleagues reported that hyperoxia reduced the level of VEGF in the lungs (34). The reduction of VEGF was correlated with a reduction in the percentage of apoptotic cells.

Another possible explanation for the decreased level of VEGF is due to proteolytic degradation. Proteolytic activity degrades VEGF in a chronic skin wound (35). Whereas plasmin and trypsin both cause VEGF degradation, thrombin, elastase, and collagenase do not cleave VEGF (36). Because smoking and interstitial lung diseases are associated with chronic inflammation in the lungs, it is possible that these protease activities may degrade VEGF, leading to the decreased level of VEGF in BALF. In contrast, Beinert and coworkers reported that oxidant caused upregulation of VEGF in BALF in patients with lung cancer receiving chemoradiotherapy (37). In their report, increased levels of oxidized methionine indicated that these patients suffered from severe pulmonary oxidative stress, but the VEGF concentration in epithelial lining fluid was significantly elevated, suggesting that its antigeneity is not affected by ambient redox potentials.

Aging is another factor that influences the levels of VEGF in BALF, because BALF VEGF levels decline significantly with advancing age (30). In this study, age was not significantly different among the groups, and thus, age alone cannot explain the reduced level of VEGF in BALF.

In conclusion, VEGF in BALF are significantly reduced in subjects who smoke cigarettes and in patients with IPF, PF-CTD, and Sar. Reduced concentrations of VEGF may be important in the pathogenesis of lung diseases in these patients.

Received in original form March 26, 2001; accepted in final form April 12, 2002

	REFERENCES

TOP ABSTRACT INTRODUCTION METHODS RESULTS DISCUSSION REFERENCES

 

1. Senger DR, Connolly DT, Van De Water L, Feder J, Dvorak HF. Purification and NH2-terminal amino acid sequence of guinea pig tumor-secreted vascular permeability factor. Cancer Res 1990;50:1774–1778.[Abstract/Free Full Text]
2. Yeo T-K, Senger DR, Dvrrak HF, Freter L, Yeo K-T. Glycosylation is essential for efficient secretion but not for permeability-enhancing activity of vascular permeability factor (vascular endothelial growth factor). Biochem Biophys Res Commun 1991;179:1568–1575.[CrossRef][Medline]
3. Senger DR, Galli SJ, Dvorak AM, Perruzzi CA, Harvey VS, Dvorak HF. Tumor cells secret a vascular permeability factor that promotes accumulation of ascites fluid. Science 1983;219:983–985.[Abstract/Free Full Text]
4. Connolly DT, Olander JV, Heuvelman D, Nelson R, Monsell R, Siegal N, Haymore BL, Leimgruber R, Freder J. Human vascular permeability factor: isolation from U937 cells. J Biol Chem 1989;264:20017–20024.[Abstract/Free Full Text]
5. Brock TA, Dvorak HF, Senger DR. Tumor-secreted vascular permeability factor increases cytosolic Ca2+ and von Willebrand factor release in human endothelial cells. Am J Pathol 1991;138:213–221.[Abstract]
6. Clauss M, Gerlach M, Gerlach H, Brett J, Wang F, Familletti PC, Pan Y-CE, Olander JV, Connolly DT, Stern D. Vascular permeability factor: a tumor-derived polypeptide that induces endothelial cell and monocyte procoagulant activity, and promote monocyte migration. J Exp Med 1990;172:1535–1545.[Abstract/Free Full Text]
7. Leung DW, Cachianes G, Kuang W-J, Goeddel DV, Ferrara N. Vascular endothelial growth factor is a secreted angiogenic mitogen. Science 1989;246:1306–1309.[Abstract/Free Full Text]
8. Conn G, Bayne ML, Soderman DD, Kwok PW, Sullivan KA, Palisi TM, Hope DA, Thomas KA. Amino acid and cDNA sequence of a vascular endothelial cell mitogen that is homologous to platelet-derived growth factor. Proc Natl Acad Sci USA 1990;87:2628–2632.[Abstract/Free Full Text]
9. Koch AH, Harlow LA, Haines GK, Amento EP, Unemori EU, Wong WL, Pope RM, Ferrara N. Vascular endothelial growth factor: a cytokine modulating endothelial function in rheumatoid arthritis. J Immunol 1994;152:4149–4156.[Abstract]
10. Unemori EN, Ferrara N, Bauer EA, Amento EP. Vascular endothelial growth factor induces interstitial collagenase expression in human endothelial cells. J Cell Physiol 1992;153:557–562.[CrossRef][Medline]
11. Mandriota SJ, Segherri G, Vassali JD, Ferrara N, Wasi S, Mazzieri R, Mignatti P, Pepper MS. Vascular endothelial growth factor increases urokinase receptor expression in vascular endothelial cells. J Biol Chem 1995;270:9709–9716.[Abstract/Free Full Text]
12. Tischer E, Mitche R, Hartman T, Silva M, Gospodarowicz D, Fiddes JC, Abraham JA. The human gene for vascular endothelial growth factor: multiple protein forms are encoded through alternative splicing. J Biol Chem 1991;266:11947–11954.[Abstract/Free Full Text]
13. Houkt KA, Ferrara N, Winer J, Cachianes G, Li B, Leung DW. The vascular endothelial growth factor family: identification of a fourth molecular species and characterization of alternative splicing of RNA. Mol Endocrinol 1991;5:1806–1814.[CrossRef][Medline]
14. Breier G, Albrecht U, Sterrer S, Risau W. Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. Development 1992;114:521–532.[Abstract]
15. Houck KA, Ferrera N, Winer J, Cachianes G, Li B, Leung DW. The vascular endothelial growth factor family: identification of a fourth molecular species and characterization of alternative splicing of RNA. Mol Endocrinol 1991;5:1806–1814.
16. Thomas KA. Vascular endothelial growth factor, a potent and selective angiogenic agent. J Biol Chem 1996;271:603–606.[Free Full Text]
17. Shifren JL, Doldi N, Ferrara N, Mesiano S, Jeffe RB. In the human fetus, vascular endothelial growth factor is expressed in epithelial cells and myocytes, but not vascular endothelium-implication for mode of action. J Clin Endocrinol Metab 1994;79:316–322.[Abstract]
18. Monacci WT, Merrill MJ, Oldfield EH. Expression of vascular permeability factor/endothelial growth factor in normal rat tissue. Am J Physiol 1993;264:C995–C1002.[Abstract/Free Full Text]
19. Kasahara Y, Tuder RM, Laimute Tarasevicience-Stewart L, Le Cras TD, Abman S, Hirth PK, Waltenberger J, Voelkel N. Inhibition of VEGF receptors causes lung cell apoptosis and emphysema. J Clin Invest 2000;106:1311–1319.[Medline]
20. Nagai S, Takeuchi M, Watanabe K, Aung H, Izumi T. Smoking and interleukin-1 activity released from human alveolar macrophages in healthy subjects. Chest 1988;94:694–700.[Abstract/Free Full Text]
21. Kaner RJ, Crystal RG. Compartmentalization of vascular endothelial growth factor to the epithelial surface of the human lung. Mol Med 2001;7:240–246.[Medline]
22. Monacci WT, Merrill MJ, Oldfield EH. Expression of vascular permeability factor/vascular endothelial growth factor in normal rat tissue. Am J Physiol 1993;264:C995–C1002.
23. Berse B, Brown LF, Van De Water L, Dvorak HF, Senger DR. Vascular permeability factor (vascular endothelial growth factor) gene is expressed differently in normal tissues, macrophages, and tumors. Mol Biol Cell 1993;3:211–220.[Abstract]
24. Clarffey KP, Wilkinson WO, Spiegelman BM. Vascular endothelial growth factor: regulation by cell differentiation and activated second messenger pathways. J Biol Chem 1992;267:16317–16322.[Abstract/Free Full Text]
25. Berkman RA, Merrill MJ, Reinhold WC, Monacci WT, Saxena A, Clark WC, Robertson JT, Ali IU, Oldfield EH. Expression of the vascular permeability growth factor/vascular endothelial growth factor gene in central nervous system neoplasmas. J Clin Invest 1993;91:153–159.
26. Connolly DT, Olander JV, Heuvelman D, Nelson R, Monsell JR, Siegel N, Haymore BL, Leingruber RM, Feder J. Human vascular permeability factor: isolation from U937 cells. J Biol Chem 1989;264:20017–20024.
27. Pekala P, Marlow M, Heuvelman D, Connolly D. Regulation of hexose transport in aortic endothelial cells by vascular permeability factor and tumor necrosis factor-, but not by insulin. J Biol Chem 1990;265:18051–18054.[Abstract/Free Full Text]
28. Shaheen RM, Davis DW, Liu W, Zebrowski BK, Wilson MR, Bucane CD, McMaaahon DJ, Ellis LM. Antiangiogenic therapy targeting the tyrosine kinase inhibitor for vascular endothelial growth factor receptor inhibits the growth of colon cancer liver metastasis and induces tumor and endothelial cell apoptosis. Cancer Res 1999;59: 5412–5416.[Abstract/Free Full Text]
29. Koyama S, Sato E, Nomura H, Masubuchi T, Takamizawa A, Kubo K, Nagai S, Izumi T. Gene regulation and protein secretion of vascular endothelial growth factor from airway epithelial cells [abstract]. Am J Respir Crit Care Med 2000;161:A149.
30. Meyer KC, Cardoni A, Xiang ZZ. Vascular endothelial growth factor in bronchoalveolar lavage from normal subjects and patients with diffuse parenchymal lung disease. J Lab Clin Med 2000;135:332–338.[CrossRef][Medline]
31. Barbas-Filho JV, Ferreira MA, Sesso A, Kairalla RA, Carvalho CR, Capelozzi VL. Evidence of type II pneumocyte apoptosis in the pathogenesis of idiopathic pulmonary fibrosis (IFP)/usual interstitial pneumonia (UIP). J Clin Pathol 2001;54:132–138.[Abstract/Free Full Text]
32. Bamzet N, Francois D, Polla BS. Tobacco smoke induces mitochondrial depolarization along with cell death: effects of antioxidanta. Redox Rep 1999;4:229–236.[CrossRef][Medline]
33. Vayssier M, Banzet N, Francois D, Bellmann K, Polla BS. Tobacco smoke induces both apoptosis and necrosis in mammalian cells: different effects of HSP70. Am J Physiol 1998;275:L771–L779.[Abstract/Free Full Text]
34. Klekamp JG, Jarzecka K, Perkett EA. Exposure to hyperoxia decreases the expression of vascular endothelial growth factor and its receptors in adult rat lungs. Am J Pathol 1999;154:823–832.[Abstract/Free Full Text]
35. Lauer G, Sollberg S, Cole M, Flamme I, Sturzenbecher J, Mann K, Krieg T, Eming SA. Expression and proteolysis of vascular endothelial growth factor is increased in chronic wounds. J Invest Dermatol 2000;115:12–18.[CrossRef][Medline]
36. Keyt BA, Berleau LT, Nguyen HV, Chen H, Heinsohn H, Vandlen R, Ferrara N. The carboxyl-terminal domain (111–165) of vascular endothelial growth factor is critical for its mitogenic potency. J Biol Chem 1996;271:7788–7795.[Abstract/Free Full Text]
37. Beinert T, Binder D, Oehm C, Ziemer S, Priem F, Stuschke M, Schweigert M, Siebert G, Mergenthaler HG, Schmid P, et al. Further evidence for oxidant-induced vascular endothelial growth factor up-regulation in the bronchoalveolar lavage fluid of lung cancer patients undergoing radio-chemotherapy. J Cancer Res Clin Oncol 2000;126: 352–356.[CrossRef][Medline]

This article has been cited by other articles:

				R. J. Giordano, J. Lahdenranta, L. Zhen, U. Chukwueke, I. Petrache, R. R. Langley, I. J. Fidler, R. Pasqualini, R. M. Tuder, and W. Arap Targeted Induction of Lung Endothelial Cell Apoptosis Causes Emphysema-like Changes in the Mouse J. Biol. Chem., October 24, 2008; 283(43): 29447 - 29460. [Abstract] [Full Text] [PDF]

				I. Edirisinghe, S.-R. Yang, H. Yao, S. Rajendrasozhan, S. Caito, D. Adenuga, C. Wong, A. Rahman, R. P. Phipps, Z.-G. Jin, et al. VEGFR-2 inhibition augments cigarette smoke-induced oxidative stress and inflammatory responses leading to endothelial dysfunction FASEB J, July 1, 2008; 22(7): 2297 - 2310. [Abstract] [Full Text] [PDF]

				B. Yucesoy, V. J. Johnson, G. E. Kissling, K. Fluharty, M. L. Kashon, J. Slaven, D. Germolec, V. Vallyathan, and M. I. Luster Genetic susceptibility to progressive massive fibrosis in coal miners Eur. Respir. J., June 1, 2008; 31(6): 1177 - 1182. [Abstract] [Full Text] [PDF]

				K. Kamio, T. Sato, X. Liu, H. Sugiura, S. Togo, T. Kobayashi, S. Kawasaki, X. Wang, L. Mao, Y. Ahn, et al. Prostacyclin analogs stimulate VEGF production from human lung fibroblasts in culture Am J Physiol Lung Cell Mol Physiol, June 1, 2008; 294(6): L1226 - L1232. [Abstract] [Full Text] [PDF]

				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]

				M. V. Avdalovic, L. F. Putney, E. S. Schelegle, L. Miller, J. L. Usachenko, N. K. Tyler, C. G. Plopper, L. J. Gershwin, and D. M. Hyde Vascular Remodeling Is Airway Generation-Specific in a Primate Model of Chronic Asthma Am. J. Respir. Crit. Care Med., November 15, 2006; 174(10): 1069 - 1076. [Abstract] [Full Text] [PDF]

				E. R. Jacobs, D. Zhu, S. Gruenloh, B. Lopez, and M. Medhora VEGF-induced relaxation of pulmonary arteries is mediated by endothelial cytochrome P-450 hydroxylase Am J Physiol Lung Cell Mol Physiol, September 1, 2006; 291(3): L369 - L377. [Abstract] [Full Text] [PDF]

				A R L Medford and A B Millar Vascular endothelial growth factor (VEGF) in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS): paradox or paradigm? Thorax, July 1, 2006; 61(7): 621 - 626. [Abstract] [Full Text] [PDF]

				G. P. Fadini, M. Schiavon, M. Cantini, I. Baesso, M. Facco, M. Miorin, M. Tassinato, S. V. d. Kreutzenberg, A. Avogaro, and C. Agostini Circulating Progenitor Cells Are Reduced in Patients with Severe Lung Disease Stem Cells, July 1, 2006; 24(7): 1806 - 1813. [Abstract] [Full Text] [PDF]

				J. A. Marwick, C. S. Stevenson, J. Giddings, W. MacNee, K. Butler, I. Rahman, and P. A. Kirkham Cigarette smoke disrupts VEGF165-VEGFR-2 receptor signaling complex in rat lungs and patients with COPD: morphological impact of VEGFR-2 inhibition Am J Physiol Lung Cell Mol Physiol, May 1, 2006; 290(5): L897 - L908. [Abstract] [Full Text] [PDF]

				K. M. Oltmanns, H. Gehring, S. Rudolf, B. Schultes, C. Hackenberg, U. Schweiger, J. Born, H. L. Fehm, and A. Peters Acute hypoxia decreases plasma VEGF concentration in healthy humans Am J Physiol Endocrinol Metab, March 1, 2006; 290(3): E434 - E439. [Abstract] [Full Text] [PDF]

				N. F. Voelkel, R. W. Vandivier, and R. M. Tuder Vascular endothelial growth factor in the lung Am J Physiol Lung Cell Mol Physiol, February 1, 2006; 290(2): L209 - L221. [Abstract] [Full Text] [PDF]

				L. K. A. Lundblad, J. Thompson-Figueroa, T. Leclair, M. J. Sullivan, M. E. Poynter, C. G. Irvin, and J. H. T. Bates Am. J. Respir. Crit. Care Med., December 15, 2005; 172(12): 1605b - 1606. [Full Text] [PDF]

				L. B. Ware, R. J. Kaner, R. G. Crystal, R. Schane, N. N. Trivedi, D. McAuley, and M. A. Matthay VEGF levels in the alveolar compartment do not distinguish between ARDS and hydrostatic pulmonary oedema Eur. Respir. J., July 1, 2005; 26(1): 101 - 105. [Abstract] [Full Text] [PDF]

				K. Nagai, T. Betsuyaku, Y. Ito, Y. Nasuhara, and M. Nishimura Decrease of vascular endothelial growth factor in macrophages from long-term smokers Eur. Respir. J., April 1, 2005; 25(4): 626 - 633. [Abstract] [Full Text] [PDF]

				T. Higenbottam Pulmonary Hypertension and Chronic Obstructive Pulmonary Disease: A Case for Treatment Proceedings of the ATS, April 1, 2005; 2(1): 12 - 19. [Abstract] [Full Text] [PDF]

				A R L Medford, I Saleem, P E Brenchley, N R Simler, and J J Egan VEGF in idiopathic ILD Thorax, April 1, 2005; 60(4): 353 - 354. [Full Text] [PDF]

				A R L Medford, L J Keen, J L Bidwell, and A B Millar Vascular endothelial growth factor gene polymorphism and acute respiratory distress syndrome Thorax, March 1, 2005; 60(3): 244 - 248. [Abstract] [Full Text] [PDF]

				G D Perkins, J Roberts, D F McAuley, L Armstrong, A Millar, F Gao, and D R Thickett Regulation of vascular endothelial growth factor bioactivity in patients with acute lung injury Thorax, February 1, 2005; 60(2): 153 - 158. [Abstract] [Full Text] [PDF]

				A G Richter, E O Maughan, G D Perkins, N Nathani, and D R Thickett VEGF levels in pulmonary fibrosis Thorax, February 1, 2005; 60(2): 171 - 171. [Full Text] [PDF]

				Y. Abadie, F. Bregeon, L. Papazian, F. Lange, B. Chailley-Heu, P. Thomas, P. Duvaldestin, S. Adnot, B. Maitre, and C. Delclaux Decreased VEGF concentration in lung tissue and vascular injury during ARDS Eur. Respir. J., January 1, 2005; 25(1): 139 - 146. [Abstract] [Full Text] [PDF]

				K. Tang, H. B. Rossiter, P. D. Wagner, and E. C. Breen Lung-targeted VEGF inactivation leads to an emphysema phenotype in mice J Appl Physiol, October 1, 2004; 97(4): 1559 - 1566. [Abstract] [Full Text] [PDF]

				G. P. Cosgrove, K. K. Brown, W. P. Schiemann, A. E. Serls, J. E. Parr, M. W. Geraci, M. I. Schwarz, C. D. Cool, and G. S. Worthen Pigment Epithelium-derived Factor in Idiopathic Pulmonary Fibrosis: A Role in Aberrant Angiogenesis Am. J. Respir. Crit. Care Med., August 1, 2004; 170(3): 242 - 251. [Abstract] [Full Text] [PDF]

				N R Simler, P E Brenchley, A W Horrocks, S M Greaves, P S Hasleton, and J J Egan Angiogenic cytokines in patients with idiopathic interstitial pneumonia Thorax, July 1, 2004; 59(7): 581 - 585. [Abstract] [Full Text] [PDF]

				N. Kaminski, J. A. Belperio, P. B. Bitterman, L. Chen, S. W. Chensue, A. M.K. Choi, S. Dacic, J. H. Dauber, R. M. du Bois, J. J. Enghild, et al. Idiopathic Pulmonary Fibrosis Am. J. Respir. Cell Mol. Biol., September 1, 2003; 29(3): S1 - 105. [Full Text] [PDF]

				R. P. Baughman and E. E. Lower The Variability of Sarcoidosis: Can We Predict It? Chest, May 1, 2003; 123(5): 1329 - 1332. [Full Text] [PDF]

				M. J. Tobin Tuberculosis, Lung Infections, Interstitial Lung Disease, and Journalology in AJRCCM 2002 Am. J. Respir. Crit. Care Med., February 1, 2003; 167(3): 345 - 355. [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Koyama, S. Articles by Izumi, T. Search for Related Content PubMed PubMed Citation Articles by Koyama, S. Articles by Izumi, T.