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J Thorac Cardiovasc Surg 2008;136:65-72
© 2008 The American Association for Thoracic Surgery

Surgery for Acquired Cardiovascular Disease

A proteomic study of the aortic media in human thoracic aortic dissection: Implication for oxidative stress

^a Department of Vascular Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China
^b Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China
^c Research Center for Proteome Analysis, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China
^d Departments of Molecular Cardiology and Nephrology, Cleveland Clinic, Ohio

Received for publication April 24, 2007; revisions received October 29, 2007; accepted for publication November 15, 2007.

^_* Address for reprints: Zaiping Jing, MD, Department of Vascular Surgery, Changhai Hospital, Second Military Medical University, No. 174 Changhai Rd, Shanghai, 200433 China. (Email: sh.vascular{at}yahoo.com.cn).

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 
Objective: The aortic media lesion is a key pathologic feature in thoracic aortic dissection. To identify key proteins in aortic media lesions that may contribute to its pathogenesis, we performed proteomic studies to find differentially expressed proteins in the media from diseased and normal thoracic aorta.

Methods: Ascending aortic segments were obtained from patients with thoracic aortic dissection (n = 8) and age-matched normal donors (n = 6). The differentially expressed proteins of their media tissues were analyzed by 2-dimensional electrophoresis and mass spectrometry, and verified by Western blotting. Oxidative stress was measured by functional assays in a larger sample size (15 patients and 10 controls).

Results: Image analysis of the protein profiles from 2-dimensional gels revealed 126 differentially expressed proteins, of which 26 were identified by mass spectrometry. Among them, extracellular superoxide dismutase, an enzyme involved in oxidative stress, was selected for further studies. Western blotting showed that extracellular superoxide dismutase expression was more than 50% lower in patient samples than in controls (P < .001). Superoxide dismutase activity was consistently decreased (P < .001) and lipid peroxidation was increased (P = .019) in patient media homogenates compared with that in controls.

Conclusion: Our results indicate that protein expression profiles in the aortic media from thoracic aortic dissection differ significantly from that of controls, which may provide important insights into the disease mechanisms. This study also suggests that increased oxidative stress may play an important role in the disease.

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 
Thoracic aortic dissection (TAD) is one of the most catastrophic aortic diseases. It is characterized by the separation of thoracic aortic wall layers by extraluminal blood that usually enters the vessel wall via an intimal tear.¹ Mechanisms weakening the aortic wall can induce aortic dilatation, aneurysm formation, and even rupture.¹ The development of endovascular techniques has improved the diagnosis and treatment of TAD. To date, however, the mortality associated with aortic rupture remains high. Many patients with TAD die of aortic rupture before reaching hospitals. Thus, it is important to develop strategies to prevent this catastrophic disease. Many studies have been performed to understand the pathogenesis of TAD. Most of them focus on the genetic heterogeneity, clinical pathology, and hemodynamics of TAD,^2,3 which have provided important insights into the disease mechanisms.

Proteomics using 2-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS)⁴ has recently been applied to study cancer⁵ and cardiovascular diseases.⁶ Comparative proteomics (ie, diseased vs normal tissue) has been used to study atherosclerosis, including coronary and carotid atherosclerotic plaques and the smooth muscle cell (SMC) secretome.^7-9 McGregor and colleagues¹⁰ presented a protein profile map of human saphenous vein medial smooth muscle by 2-DE and identified 129 differentially expressed proteins. To date, comparative proteomic profiles of the aorta from patients with TAD have not been reported. Such data will help to identify key proteins involved in TAD and elucidate their functions in the disease.

In this study, we initiated a proteomic analysis of aortic tissues from patients with TAD. Because cellular composition in the aortic wall is heterogeneous, a proteomic analysis of the entire aortic wall will be technically challenging. For instance, intimal atherosclerotic lesions are present in most aortas from patients with TAD, whereas such lesions are less common in normal thoracic aortas (NAs), making comparative studies difficult. To circumvent this problem, we focused on the aortic media, which is more homogenous in its cellular composition.

By using 2-DE and MS, we produced a first map of human thoracic aortic media protein profiles and analyzed differentially expressed proteins between the TAD and NA groups. A total of 126 differentially expressed proteins were identified, of which 26 proteins were analyzed by MS. One of them, extracellular superoxide dismutase (EC-SOD), was selected for in-depth studies.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 
Proteomic Analysis
Aortic samples
The study was approved by the ethics committee of our hospital. All patients gave informed consent. Ascending aorta segments were collected from patients with TAD undergoing surgical repair (8 male, mean age: 47.5 ± 6.9 years). NAs were from organ donors (6 male, mean age: 39.5 ± 7.2 years). No significant difference in age was found between the TAD and NA groups (P > .05). A detailed description of aortic samples is supplemented online (Table E1). All samples used in this study were prepared in parallel on the basis of a published method.¹⁰ Briefly, media tissues were cut into small pieces and stored at –80°C until protein extraction. Some tissues were preserved by paraffin imbedding for immunohistochemistry validation using monoclonal antibody specific for SMC marker (alpha smooth muscle actin) and endothelial cell marker (CD-31). Hematoxylin and eosin and Victoria blue staining for elastic and collagen fibers were done for all tissue samples using standard procedures.

Preparation of tissue samples
Tissue samples were prepared according to the methods described by Weekes and colleagues¹¹ and McGregor and colleagues.¹⁰ Frozen aortic tissues (10 mg) were ground using a grinding tube with a matching pestle in an iced bath. Resulting powders were homogenized in 100 µL lysis buffer (9 mol/L urea, 4% CHAPS, 65 mmol/L dithiothreitol, and 1 mmol/L phenylmethylsulfonyl fluoride) at 4°C for 5 minutes and then centrifuged at 4°C at 14,000 rpm for 1 hour. The supernatant was collected, and protein concentrations were determined using a modified method described by Bradford.¹² Supernatant samples were divided into 50-µL aliquots and stored at –80°C.

Two-dimensional gel electrophoresis
2-DE was performed according to the methods described by Weekes and colleagues¹¹ and Yu and colleagues.¹³ The details of these methods are provided in supplemented documents online (Appendix E1).

Image analysis
Images of stained gels were acquired using a scanner (GS-800 densitometry, Bio-Rad, Richmond, Calif) and analyzed with PDQuest 7.2.0 2-dimensional image analysis software (Bio-Rad).¹⁴ Spot densities were determined following normalization based on total spot volumes on the gel. Protein spots from different gels were matched. Protein spots that had significant changes in densities (P < .05) in a consistent direction (increase or decrease) were considered to be different.

Mass spectrometry
In-gel tryptic digestion and MS identification were carried out according to published methods.^13,14 The peptide mass spectrum for each sample was obtained with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight MS (Bruker, Daltonics Autoflex Billerica, MA). The obtained peptide masses were searched against the National Center for Biotechnology Information protein database using the Mascot program (Matrix Science, Boston, Mass).

Western Blotting
Western blotting analysis of aortic protein extracts was performed to validate EC-SOD expression with standard procedures. A rabbit anti–EC-SOD polyclonal antibody (Alpha Diagnostic International, San Antonio, TX), and horseradish peroxidase-labeled secondary antibody (Santa Cruz Biochemicals, Santa Cruz, Calif) were used in these studies.

Assays for Oxidative Stress
Superoxide dismutase (SOD) activity, lipid peroxidation, and catalase activity in aortic media tissues were determined. In addition to the samples used in 2-DE, we included additional samples from other patients to increase sample size and to confirm our findings in independent patients. There was no significant difference in age between the TAD group (13 men and 2 women, mean age: 47.4 ± 9.2 years) and NA group (9 men and 1 woman, mean age: 40.9 ± 6.7 years) (P > .05). A detailed description of these samples is supplemented online (Table E1). SOD activity in aortic media homogenates was assayed on the basis of a published method that measures the inhibitory rate of xanthine oxidase-mediated reduction of cytochrome c (pH 7.4).¹⁵ Cyanide (3 mmol/L) was used to distinguish between the cyanide-sensitive isozymes Cu/Zn-SOD and EC-SOD from the cyanide-resistant manganese-SOD. Lipid peroxidation was assayed by measuring the levels of malondialdehyde (MDA) reacted with thiobarbituric acid at 535 nm.¹⁶ MDA concentrations were expressed as nanomoles per milliliter. Catalase activity was determined by monitoring the breakdown of hydrogen peroxide catalyzed by the catalase enzyme,¹⁷ and the results were expressed as units per milligram of protein.

Statistical Analysis
Statistical analyses were performed using the Statistical Package for the Social Sciences 15.0 (SPSS Inc, Chicago, IL.). Continuous and categoric variables were compared by t test and chi-square/Fisher exact test, respectively. Descriptions of continuous variables were presented as mean ± standard deviation.

	Results

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 
Histopathology of Thoracic Aortic Dissection
The ascending aortas from patients with TAD were enlarged and dissected. In most samples examined, large amounts of blood clots were found in false lumens ( Figure 1, A). Microscopic examination of hematoxylin and eosin- and Victoria blue-stained tissues (Figure 1, B and C) showed that the dissection spread along the laminate of the media between the middle and outer third layers. The aortic wall specimen had elastic fiber loss and fragmentation in the media. Medial tissue identity was confirmed by positive immunostaining of an anti-alpha smooth muscle actin antibody (Figure 1, D) and negative staining of an anti-CD31 antibody (data not shown).

View larger version (129K): [in this window] [in a new window]   Figure 1. Histopathology of TAD. An image of an ascending aorta from a patient with TAD showing an enlarged and dissected aorta with large amounts of blood clots in the false lumen (A). Hematoxylin and eosin staining of TAD aortic tissue showing the dissected layer in the middle media (B). Fragmentation and rigidity of elastic fiber (blue) in TAD by Victoria blue staining (C). Verification of aortic media by immunostaining of alpha smooth muscle actin (brown) in the media (D).

View larger version (72K): [in this window] [in a new window]   Figure 2. 2-DE images of protein profiles of the aortic media. Representative 2-DE images of aortic media protein from a normal control (A) and patient with TAD (B). Protein spots identified by MS were marked with numbers. Enlarged pictures of silver-stained gels highlight the significant differences between samples from patients and controls (C).

View larger version (17K): [in this window] [in a new window]   Figure 3. Peptide mass fingerprinting and tandem mass spectrophotometry spectrum of spot 15. Peptide mass fingerprinting spectrum of spot 15 by matrix-assisted laser desorption/ionization time-of-flight-MS (A). Three peptides (red) were selected for tandem mass spectrophotometry identification. Tandem mass spectrophotometry spectrum of spot 15 (B) was from 1418 m/z (A, gray circle). Spot 15 was identified to be EC-SOD.

Western Blotting Verification of Extracellular Superoxide Dismutase Expression
Among the differentially expressed proteins identified, one protein, EC-SOD, is of particular interest because of its role in oxidative stress. We used Western blotting to verify its expression in the tissue samples used in the 2-DE experiments. EC-SOD protein was found in samples from both NA and TAD groups ( Figure 4, A). The level of protein expression was significantly lower in the TAD group than in the NA group (0.43 ± 0.04 vs 1.01 ± 0.03 relative amount of protein, respectively; P < .001) (Figure 4, B).

View larger version (25K): [in this window] [in a new window]   Figure 4. Western blotting verification for EC-SOD protein expression. EC-SOD and the internal control protein (beta-actin) were detected in aortic samples from the NA and TAD groups (A). Quantitative analysis of EC-SOD expression in TAD (n = 8) and NA (n = 6) groups (B). * P < .05. EC-SOD, Extracellular superoxide dismutase; NA, normal thoracic aorta; TAD, thoracic aortic dissection.

View larger version (14K): [in this window] [in a new window]   Figure 5. MDA levels, catalase activity, and SOD activity in aortic samples. Increased MDA in the TAD group compared with that in the NA group (A). Catalase activity was lower in the TAD group than in the NA group, but the difference was not statistically significant (B). Total SOD activity and activities of cyanide-sensitive SOD (Cu/Zn-SOD plus EC-SOD) were also decreased in the TAD group compared with the NA group (C). * P < .05. MDA, Malondialdehyde; NA, normal thoracic aorta; TAD, thoracic aortic dissection; EC-SOD, extracellular superoxide dismutase; Mn-SOD, manganese-superoxide dismutase; SOD, superoxide dismutase.

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

Oxidative stress plays a key role in the pathogenesis of vascular diseases.¹⁸ EC-SOD is a major form of SOD in the vessel wall and represents an important enzymatic antioxidant defense system.¹⁵ Decreased EC-SOD expression has been found to be associated with increased vascular oxidative stress.¹⁸ We found that EC-SOD expression was markedly reduced in the media samples from patients with TAD in this proteomic study (Figure 2, spots 15 and 16). Therefore, we selected this protein for further validation studies.

We examined EC-SOD protein expression in aortic media tissues by Western blotting and confirmed its lower expression in TAD samples (Figure 4). This finding was supported in additional functional assays. We showed that total SOD activity and Cu/Zn-SOD activities decreased markedly in TAD samples compared with that in normal controls (Figure 5, C). In addition, the MDA level, an indicator of lipid peroxidation,¹⁶ was increased significantly in TAD tissues (Figure 5, A). Our results indicate that oxidative stress is significantly increased in the aortic wall in patients with TAD. The data suggest that increased oxidative stress may be an important contributing factor in the pathogenesis of TAD. However, we cannot rule out at this time that the observed EC-SOC expression change is partially due to the inflammatory reaction in the TAD tissue. It is equally possible that the aorta in patients with TAD reacts poorly to existing oxidative stress because of the decreased expression of free radical scavengers, such as SOD, which may eventually lead to detrimental changes in its structure. These possibilities need to be tested in future experiments.

In addition to EC-SOD, we found other proteins that were differentially expressed between the TAD and NA groups. Among them, destrin (spot 1) is a member of the actin depolymerizing factor family and regulates actin cytoskeleton in various eukaryotes.¹⁹ Smooth muscle 22-alpha, represented by spots 2 to 5 for its isoforms or modifications of the protein, is a SMC-specific actin binding protein that is considered as an earliest contractile marker of differentiated SMCs.²⁰ Feil and colleagues²¹ showed that smooth muscle 22 alpha–modulated vascular SMCs phenotype during vascular remodeling. Skeletal muscle LIM-protein FHL1 (spot 19) and ALP (spot 20) are involved in cytoskeletal integrity²² and reorganization.²³ The altered expression of these cytoskeletal proteins in the aortic media of patients with TAD indicates that cytoskeletal reorganization may be impaired in the aorta, suggesting a possible mechanism of defective aortic remodeling in the disease.

We also found 2 isoforms of HSP27 (spots 13 and 14) in the aortic media of patients with TAD. These proteins are similar to 2 distinct HSP27 isoforms identified in human venous media by McGregor and colleagues.¹⁰ HSP27 is likely to play a role in the actin filament remodeling during SMC migration and contractions.²⁴ It has been suggested that HSP27 increases the resistance against oxidative stress-induced actin fragmentation.²⁵ Therefore, HSP27 may play a role in the remodeling process of TAD, partially by regulating SMC functions.

Another interesting finding was the decreased osteoglycin protein (spots 17 and 18) in patients with TAD. Previous studies on osteoglycin expression in rat SMCs and carotid arteries, as well as human coronary arteries, have suggested that this protein is a new marker for differentiated SMCs and may be an essential component of the normal vascular matrix.²⁶ The decreased osteoglycin expression in patients with TAD suggests that SMC phenotypic change and vascular matrix remodeling may occur in the aortic media of TAD.

	Conclusions

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 
Our proteomic analysis of the aortic media from the TAD and NA groups identified a number of differentially expressed proteins. In particular, we found expression pattern changes in EC-SOD, osteoglycin, and some cytoskeletal proteins in tissues from patients with TAD. These new findings suggest that increased oxidative stress and impaired vessel wall remodeling may be possible molecular mechanisms in the pathogenesis of TAD. We recognize that the patient sample size in this study was small and that protein spots identified by computer-based statistical algorithm need to be verified experimentally. At this time, it is unclear whether the protein expression changes observed are the cause or effect of the dissection. Thus, additional mechanistic studies are needed to define the role of the identified proteins in TAD. Our results should encourage further characterization of the differentially expressed proteins, which may lead to new hypotheses. Together, a better understanding of the molecular mechanisms underlying TAD should help to design more effective strategies to diagnose and treat this life-threatening disease.

The use of the facilities of the Research Center for Proteome Analysis of the Shanghai Institute of Biochemistry is gratefully acknowledged. We thank Dr Jibin Xu (Department of Thoracic Surgery, Changhai Hospital) for providing aortic samples and Dr Matthew Eagleton (Department of Vascular Surgery, Cleveland Clinic) for critical reading of this article.

	Appendix E1

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 
The first dimension immobilized pH gradients Dalt (IPG-Dalt) 2-dimensional gel electrophoresis was run on an IPGphor isoelectric focusing (IEF) system. IEF was performed using 130-mm, immobilized, nonlinear pH gradient strips (IPG DryStrips; GE Healthcare, Amershan- Biosciences, Piscataway, NJ) of pH 3 to 10. A total protein load of 100 µg per IPG DryStrip was mixed with a rehydration solution containing 8 mol/L urea, 2% CHAPS, 0.5% IPG buffer, pH 3–10 L, 18 mmol/L dithiothreitol (DTT), and a trace of bromophenol blue, to a total volume of 250 µL, and rehydrated overnight in a re-swelling tray. After rehydration for 12 hours, IEF was conducted automatically at low voltages (500–1000V) during the first 2 hours and then continued with a maximum setting of 8000V to reach a total of 40 kVh for analytic runs or 65 to 70 kVh for preparative runs at 20°C. After IEF separation, the strips were equilibrated in 6 mol/L urea containing 30% (v/v) glycerol, 2% (w/v) sodium dodecylsulfate, and 0.01% (w/v) bromphenol blue, with the addition of 1% (w/v) DTT for 15 minutes, followed by the same buffer without DTT but with the addition of 2.5% (w/v) iodoacetamide for 15 minutes. Sodium dodecylsulfate–polyacrylamide gel electrophoresis was performed using 1-mm–thick, 12% T, 2.6% C separating polyacrylamide gels without a stacking gel using the Hoefer SE 600 (Amersham). The second dimension was carried out at a constant current of 10 mA/gel for 15 minutes and then switched to 30 mA/gel until the bromphenol dye front had reached the bottom of the gels. After electrophoresis, analytic 2-dimensional gels were fixed for a minimum of 1 hour in a methanol:acetic acid:water solution (4:1:5 v/v/v). Analytic 2-dimensional protein profiles were visualized by silver staining using the OWL Silver Staining Kit (Insight Biotechnology Ltd, Wembley, UK), and preparative 2-dimensional gels were stained using Coomassie Brilliant Blue R-350 stain for subsequent mass spectrometry analysis.

	Table E1

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Clinical characteristics of groups with normal thoracic arteries and thoracic aortic dissection

NA * (n = 6) TAD * (n = 8) NA (n = 10) TAD (n = 15) Age (y) 39.5 ± 7.2 47.5 ± 6.9 40.9 ± 6.7 47.4 ± 9.2 Male/female 6/0 8/0 13/2 9/1 Maximal diameter (cm) 2.48 ± 0.29 5.56 ± 1.17 2.36 ± 0.29 5.55 ± 1.11 Atherosclerosis 2 (33.3%) 7 (87.5%) 3 (33.3%) 14 (93.3%) Hypertension 0 (0.0%) 7 (87.5%) 0 (0.0%) 13 (86.6%) Bicuspid aortic valve 0 (0.0%) 0 (0.0%) 0 (0.0%) 0 (0.0%) Marfan syndrome 0 (0.0%) 0 (0.0%) 0 (0.0%) 0 (0.0%) DeBakey type I aortic dissection — 8 (100%) — 15 (100%) * Sample groups for proteomic analysis. Sample groups for oxidative stress assay (including the samples in *groups).

NA, Normal thoracic aorta; TAD, thoracic aortic dissection.

	Table E2

Top Abstract Introduction Materials and Methods Results Discussion Conclusions Appendix E1 Table E1 Table E2 References

 

Protein sequences identified by matrix-assisted laser desorption/ionization time-of-flight and time-of-flight mass spectrometry

N Protein identity Peptide matches Sequences Sequence coverage (%) 1 Destrin 2 R.YALYDASFETK.E 17 K.HFVGMLPEKDCR.Y 3 Smooth muscle 22-alpha/transgelin 3 R.LGFQVWLK.N 19 K.TDMFQTVDLFEGK.D R.LVEWIIVQCGPDVGRPDR.G 11 Actin, alpha 2, smooth muscle, aorta 3 R.AVFPSIVGRPR.H 26 K.IWHHSFYNELR.V R.VAPEEHPTLLTEAPLNPK.A 13 Heat shock protein 27 3 R.RVPFSLLR.G 17 R.LFDQAFGLPR.L R.VSLDVNHFAPDELTVK.T 15 Superoxide dismutase 3, extracellular 3 K.VTEIWQEVMQR.R 17 R.AGLAASLAGPHSIVGR.A R.LACCVVGVCGPGLWER.Q 17 Osteoglycin 4 K.DFADIPNLR.R 19 R.IEEIRLEGNPIVLGK.H K.LSLLEELSLAENQLLK.L R.RLDFTGNLIEDIEDGTFSK.L 20 Alpha-actinin-2–associated LIM protein 4 R.LSGGIDFNQPLVITR.I 23 K.INLESEPQEFKPIGTAHNR.R R.GLIPSSPQNEPTASVPPESDVYR.M K.GYFFIEGELYCETHAR.A

	Footnotes

 
This work was supported by the National Natural Science Foundation of China (30600599).

	References

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1. Nienaber CA, Eagle KA. Aortic dissection: new frontiers in diagnosis and management. part I: from etiology to diagnostic strategies. Circulation 2003;108:628-635.[Free Full Text]
   
2. Mészáros I, Mórocz J, Szlávi J, Schmidt J, Tornóci L, Nagy L, et al. Epidemiology and clinicopathology of aortic dissection. Chest 2000;117:1271-1278.[Medline]
   
3. Guo D, Hasham S, Kuang SQ, Vaughan CJ, Boerwinkle E, Chen H, et al. Familial thoracic aortic aneurysms and dissections: genetic heterogeneity with a major locus mapping to 5q13-14. Circulation 2001;103:2461-2468.[Abstract/Free Full Text]
   
4. Blackstock WP, Weir MP. Proteomics: quantitative and physical mapping of cellular proteins. Trends Biotechnol 1999;17:121-127.[Medline]
   
5. Alaiya A, Al-Mohanna M, Linder S. Clinical cancer proteomics: promises and pitfalls. J Proteome Res 2005;4:1213-1222.[Medline]
   
6. Matt P, Carrel T, White M, Lefkovits I, VanEyk J. Proteomics in cardiovascular surgery. J Thorac Cardiovasc Surg 2007;133:210-214.[Abstract/Free Full Text]
   
7. Duran MC, Mas S, Martin-Ventura JL, Meilhac O, Michel JB, Gallego-Delgado J, et al. Proteomic analysis of human vessels: application to atherosclerotic plaques. Proteomics 2003;3:973-978.[Medline]
   
8. You SA, Archacki SR, Angheloiu G, Moravec CS, Rao S, Kinter M, et al. Proteomic approach to coronary atherosclerosis shows ferritin light chain as a significant marker: evidence consistent with iron hypothesis in atherosclerosis. Physiol Genomics 2003;13:25-30.[Abstract/Free Full Text]
   
9. Dupont A, Corseaux D, Dekeyzer O, Drobecq H, Guihot AL, Susen S, et al. The proteome and secretome of human arterial smooth muscle cells. Proteomics 2005;5:585-596.[Medline]
   
10. McGregor E, Kempster L, Wait R, Welson SY, Gosling M, Dunn MJ, et al. Identification and mapping of human saphenous vein medial smooth muscle proteins by two-dimensional polyacrylamide gel electrophoresis. Proteomics 2001;1:1405-1414.[Medline]
    
11. Weekes J, Wheeler CH, Yan JX, Weil J, Eschenhagen T, Scholtysik G, et al. Bovine dilated cardiomyopathy: proteomic analysis of an animal model of human dilated cardiomyopathy. Electrophoresis 1999;20:898-906.[Medline]
    
12. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-254.[Medline]
    
13. Yu LR, Zeng R, Shao XX, Wang N, Xu YH, Xia QC. Identification of differentially expressed proteins between human hepatoma and normal liver cell lines by two-dimensional electrophoresis and liquid chromatography-ion trap mass spectrometry. Electrophoresis 2000;21:3058-3068.[Medline]
    
14. Jin X, Li X, Wang LS, Shi JZ, Zheng Y, Chen WL, et al. Differential protein expression in hypertrophic heart with and without hypertension in spontaneously hypertensive rats. Proteomics 2006;6:1948-1956.[Medline]
    
15. Stralin P, Karlsson K, Johansson BO, Marklund SL. The interstitium of the human arterial wall contains very large amounts of extracellular superoxide dismutase. Arterioscler Thromb Vasc Biol 1995;15:2032-2036.[Abstract/Free Full Text]
    
16. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351-358.[Medline]
    
17. Yasmineh WG, Kaur TP, Blazar BR, Theologides A. Serum catalase as marker of graft-versus-host disease in allogenic bone marrow transplant recipients: pilot study. Clin Chem 1995;41:1574-1580.[Abstract/Free Full Text]
    
18. Landmesser U, Spiekermann S. Vascular oxidative stress and endothelial dysfunction in patients with chronic heart failure: role of xanthine-oxidase and extracellular superoxide dismutase. Circulation 2002;106:3073-3078.[Abstract/Free Full Text]
    
19. Yahara I, Aizawa H, Moriyama K, Iida K, Yonezawa N, Nishida E, et al. A role of cofilin/destrin in reorganization of actin cytoskeleton in response to stresses and cell stimuli. Cell Struct Funct 1996;21:421-424.[Medline]
    
20. Gimona M, Sparrow MP, Strasser P, Herzog M, Small JV. Calponin and SM22 isoforms in avian and mammalian smooth muscle. Absence of phosphorylation in vivo. Eur J Biochem 1992;205:1067-1075.[Medline]
    
21. Feil S, Hofmann F, Feil R. SM22 alpha modulates vascular smooth muscle cell phenotype during atherogenesis. Circ Res 2004;94:863-865.[Abstract/Free Full Text]
    
22. McGrath MJ, Mitchell CA, Coghill ID, Robinson PA, Brown S. Skeletal muscle LIM protein 1 (SLIM1/FHL1) induces 5β1-integrin-dependent myocyte elongation. Am J Physiol Cell Physiol 2003;285:1513-1526.
    
23. Kiwon JO, Rutten B, Bunn RC, Bredt DS. Actinin-associated LIM protein-deficient mice maintain normal development and structure of skeletal muscle. Mol Cell Biol 2001;21:1682-1687.[Abstract/Free Full Text]
    
24. An SS, Fabry B, Mellema M, Bursac P, Gerthoffer WT, Kayyali US, et al. Role of heat shock protein 27 in cytoskeletal remodeling of the airway smooth muscle cell. J Appl Physiol 2004;96:1701-1713.[Abstract/Free Full Text]
    
25. McGregor E, Kempster L, Wait R, Gosling M, Dunn MJ, Powell JT. F-actin capping (CapZ) and other contractile saphenous vein smooth muscle proteins are altered by hemodynamic stress: a proteomic approach. Mol Cell Proteomics 2004;3:115-124.[Abstract/Free Full Text]
    
26. Shanahan CM, Cary NR, Osbourn LK, Weissberg PL. Identification of osteoglycin as a component of the vascular matrix: differential expression by vascular smooth muscle cells during neointima formation and in atherosclerotic plaques. Arterioscler Thromb Vasc Biol 1997;17:2437-2447.[Abstract/Free Full Text]
    

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