|Year : 2018 | Volume
| Issue : 3 | Page : 295-302
Diagnostic value of 8-isoprostane and transforming growth factor-β in bronchial asthma patients
Amgad Frahat1, Yousef Mansour1, Ayman Eldib1, Doaa Alsed2
1 Chest Department, Faculty of Medicine, Tanta University, Tanta, Egypt
2 Tanta Chest Hospital, Tanta, Egypt
|Date of Submission||20-Feb-2017|
|Date of Acceptance||22-Oct-2017|
|Date of Web Publication||20-Aug-2018|
Source of Support: None, Conflict of Interest: None
Background Asthma is an airway inflammatory disease with functional and structural changes, leading to bronchial hyperreponsiveness and airflow obstruction. 8-Isoprostane (8-iso-PGF2α) is considered a marker of oxidative stress specific to lipid peroxidation, transforming growth factor β1 (TGFβ1) as an important fibrogenic and immunomodulatory factor known to induce structural changes associated with asthma.
Objective We aimed to study the diagnostic value of 8-iso-PGF2α and TGFβ1 in asthmatic patients.
Patients and methods Samples of serum and bronchoalveolar lavage fluid from 40 asthmatic patients (20 moderate and 20 severe) and 10 healthy volunteers were assessed for their levels of 8-iso-PGF2α and TGFβ1.
Results Bronchoalveolar lavage 8-iso-PGF2α and TGFβ1 was higher in asthmatic patients. It was significantly increased with increased asthma severity.
Conclusion Increased levels of TGFβ1 and 8-iso-PGF2α is associated with disease severity. However, there is need for continued exploration on the mechanisms responsible for these structural changes.
Keywords: airflow obstruction, asthma, biomarker, hyperreponsiveness, inflammation, 8-iso-prostaglandin F2α, transforming growth factor β1
|How to cite this article:|
Frahat A, Mansour Y, Eldib A, Alsed D. Diagnostic value of 8-isoprostane and transforming growth factor-β in bronchial asthma patients. Egypt J Bronchol 2018;12:295-302
| Introduction|| |
Asthma is a disease characterized by recurrent attacks of dyspnea and wheezing, which vary in frequency and severity from person to person. Symptoms may occur several times in a day or week in affected patients, and for some people become worse at night or during physical activity. During an asthma attack, the bronchial mucosa swell, causing the airways to narrow and reduce airflow. Recurrent asthma symptoms frequently cause sleeplessness, daytime fatigue, reduced activity levels, and school and work absenteeism .
Isoprostanes are prostaglandin-like compounds that are produced by free radical-mediated peroxidation of polyunsaturated fatty acids. There is direct evidence showing that F2-isoprostanes can be utilized as a marker of lipid peroxidation due to the mechanism of their formation, chemical stability. An altered generation of F2-isoprostanes has been found in a variety of diseases associated with oxidative stress such as bronchial asthma .
Transforming growth factor-β (TGFβ) is an important fibrogenic and immunomodulatory factor known to induce structural changes associated with asthma. TGFβ is produced in the airways by inflammatory cells infiltrated in the bronchial mucosa, as well as by structural cells of the airway wall including fibroblasts, epithelial, endothelial, and smooth muscle cells .
TGFβ is increased in asthmatic airways and cells, together with evidence of increased TGFβ signaling ,. Levels of TGFβ have been shown as a potential biomarker for asthma  as well as a therapeutic target for the modulation of airway remodeling in asthma .
| Aim|| |
To evaluate the diagnostic value of 8-isoprostane (8-iso-PGF2α) and TGFβ1 in bronchial asthma patients.
| Patients and methods|| |
This study was carried out at the Chest Department of Tanta University Hospital on 40 asthmatic patients and 10 control patients collected from the outpatient clinics starting from October 2014 to September 2015. They were classified into three groups. Group Ι included 10 healthy individuals as control. Group ΙΙ included 20 patients with moderate persistent asthma. Group ΙΙΙ included 20 patients with severe persistent asthma.
No smoking within the past 5 years and no current ailments other lung disease. Nighttime symptoms more than once per week but not nightly and FEV1 more than 60% but less than 80% in patients with moderate persistent asthma and nightly symptoms and FEV1 of less than 60% in severe persistent asthma. All patients were subjected to: thorough history taking, complete physical examination, chest radiography to exclude any other lung diseases, complete blood picture, spirometry to classify our patients including forced expiratory volume in the first second (FEV1), FEV1/percent of forced expiratory volume in the first second (FVC), forced vital capacity (FVC), peek expiratory flow rate (PEFR) before and after B2 agonist inhalation to differentiate asthma from chronic obstructive pulmonary disease, serum level of 8-iso-PGF2α and TGFβ1, flexible fiberoptic bronchoscope, with bronchoalveolar lavage (BVL) was collected for the estimation of 8-iso-PGF2α and TGFβ1.
Chest diseases other than bronchial asthma, cardiac patients, mechanically ventilated patients with a PaO2 of less than 70 mmHg, thrombocytopenia with platelets of less than 100 000/ml, patients with psychological impairment, patients receiving immunosuppressive treatment, cancer patients, hepatic cirrhosis, chronic renal failure, autoimmune or connective tissue disorders, and pregnant women.
Statistical presentation and analysis of the present study was conducted, using the mean, SD, linear correlation coefficient (r); χ2, analysis of variance tests was used for comparison among different times in the same group in quantitative data by SPSS, version 17 (SPSS Inc., Chicago, IL USA). A P value of less than 0.001 is considered highly significant.
| Results|| |
This work was carried out on 40 asthmatic patients and 10 control patients classified into three groups. Group I: 10 nonsmoking healthy volunteers (control) [seven (70%) men and three (30%) women], their ages ranged from 24 to 55 (38±8.16) years. Group II: included 20 nonsmoking, moderate intermittent asthmatic patients (eight men 40% and 12 women 60%), their ages ranged from 24 to 55 (39.35±8.23) years. Group III: included 20 nonsmoking severe persistent asthmatic patients [11 (55%) men and nine (45%) women]. Their ages ranged from 30 to 48 (37.4±5.538). On comparing the values of age and sex, they were matched with no significant difference between the three studied groups (Tables 1 and 2).
|Table 2 Statistical comparison of the percentage of sex in the studied groups|
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Bronchoalveolar lavage fluid and serum levels of 8-isoprostane (ng/ml)
The mean±SD values of 8-iso-PGF2α in BAL were 24.9±1.449, 61.25±6.455, and 89.2±9.55 ng/ml and in serum were 56.4±8.5, 467.7±89.258, and 915±66.132 ng/ml in groups I, II, and III respectively, with significant increase in groups II and III compared with group I and in group III compared with group II (P<0.001) ([Table 3] and [Table 4] and [Figure 1] and [Figure 2]).
|Table 3 Statistical comparison of range, mean±SD values of bronchoalveolar lavage fluid levels of 8-isoprostane (ng/ml) in the three studied groups|
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|Table 4 Statistical comparison of range, mean±SD values of serum fluid levels of 8-isoprostane (ng/ml) in the three studied groups|
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|Figure 1 Statistical comparison of range, mean±SD value of bronchoalveolar lavage fluid levels of 8-isoprostane (ng/ml) in the three studied groups.|
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|Figure 2 Statistical comparison of range, mean±SD values of serum fluid levels of 8-isoprostane α (ng/ml) in the three studied groups.|
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Bronchoalveolar lavage fluid and serum levels of transforming growth factor β1 (ng/ml)
The mean±SD values of TGFβ1 in BAL were 5.5±0.471, 30.1±2.1 and 43.35±5.8 ng/ml and in serum were 0.25±0.012, 31.40±3.858, and 43.1±3.447 ng/ml in groups I, II, and III, respectively, with significant increase in groups II and III compared with group I and in group III compared with group II (P<0.001) (Tables 5 and 6 and [Figure 3] and [Figure 4]).
|Table 6 Statistical comparison of range, mean±SD values of serum levels of transforming growth factor β1 (ng/ml) in the three studied groups|
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|Figure 3 Statistical comparison of range, mean±SD values of bronchoalveolar lavage fluid levels of transforming growth factor β1 (ng/ml) in the three studied groups.|
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|Figure 4 Statistical comparison of range, mean±SD values of serum levels of transforming growth factor β1 (ng/ml) in the three studied groups.|
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Correlations of 8-isoprostane concentration (ng/ml) in bronchoalveolar lavage and serum and spirometric values
There was significant negative correlation between 8-iso-PGF2α concentration in BAL and FEV1 (percentage of predicted) (r=−0.790 and P<0.001), FVC (percentage of predicted) (r=−0.529 and P≤0.001), PEFR (percentage of predicted) (r=−0.682 and P≤0.001) as well as there was significant negative correlation between serum 8-iso-PGF2α concentration and percentage of predicted FEV1 (r=−0.893 and P<0.001), FVC (r=−0.667 and P<0.001), and PEFR (r=−0.785 and P≤0.001) ([Figure 5] and [Figure 6]).
|Figure 5 Correlation of 8-isoprostane concentration (ng/ml) in bronchoalveolar lavage and FVE1% of predicted.|
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|Figure 6 Correlation of 8-isoprostane concentration (ng/ml) in serum and FEV1% of predicted.|
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Correlations of transforming growth factor β1 concentration (ng/ml) in bronchoalveolar lavage and serum and spirometric values
There was significant negative correlation between TGFβ1 concentration in BAL and FEV1 (percentage of predicted) (r=−0.791 and P<0.001), FVC (percentage of predicted) (r=−0.579 and P≤0.001), PEFR (percentage of predicted) (r=−0.729 and P≤0.001) as well as there was significant negative correlation between serum TGFβ1 concentration and FEV1 (percentage of predicted) (r=−0.782 and P<0.001), FVC (percentage of predicted) (r=−0.576 and P<0.001), and PEFR (percentage of predicted) (r=−0.719 and P≤0.001) ([Figure 7] and [Figure 8]).
|Figure 7 Correlation of transforming growth factor β1 concentration (ng/ml) in bronchoalveolar lavage and FEV1% of predicted.|
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|Figure 8 Correlation of transforming growth factor β1 concentration (ng/ml) in serum and FEV1% of predicted.|
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| Discussion|| |
Asthma is considered one of the major causes of chronic morbidity and mortality worldwide. There is evidence that asthma prevalence has increased considerably over the last 20 years. Asthma is a chronic airway disorder characterized by interaction of airway inflammation, airway obstruction, and bronchial hyperresponsiveness, which leads to recurrent attacks of chest tightness, dyspnea, wheezing, and coughing . Airway inflammation and remodeling are the two main pathological features of asthma. It is proved that asthma is a nonspecific airway inflammatory disease in which many cells such as eosinophils, T-lymphocytes, neutrophils, epithelial cells, and other cellular components are involved . 8-iso-PGF2α is considered a marker of oxidative stress specific for lipid peroxidation, which is a stable metabolite of arachidonic acid, synthesized in vivo . Previous studies found elevation of 8-iso-PGF2α levels in exhaled breath condensate, induce sputum, and in plasma of asthmatic patients ,,. The human TGFβ1 is an important fibrogenic and immunomodulatory factor known to induce structural changes associated with asthma . It is produced in the airways by inflammatory cells infiltrated in the bronchial mucosa, as well as by structural cells of the airway wall including fibroblasts, endothelial, epithelial, and smooth muscle cells. These studies point out that the role for TGF not only as a potential biomarker but as a therapeutic target for the modulation of airway remodeling in asthma .
In asthmatic patients the TGFβ1 levels were found elevated in both bronchoalveolar lavage fluids (BVLFs) and plasma . Recently research workers’ pay considerable attention for airway remodeling in asthmatics. The structural changes observed by light microscopy are multiple. All the changes of remodeling are deviations from what is probably an optimized airways and so it should ideally be reversed by treatment. To date, there is a lack of evidence that the principal changes are indeed reversible but reversal of remodeling is a target goal of existing and future therapies .
In the present study, there was significant increase of 8-iso-PGF2α in BAL and serum in moderate and severe asthma compared with control, and in severe asthma compared with the moderate one. Kharitonov et al. , found doubling of 8-iso-PGF2α levels of mild asthmatic patients compared with healthy individuals and three times increase in patients with severe asthma, irrespective of treatment with corticosteroids. Dworski et al.  found elevation of F2-isoPGs in BALF after 24 h of allergen instillation compared with its level at baseline. The rise of the concentrations of F2-isoPGs in BALF 24 h after allergen challenge was explained by its generation directly in the asthmatic airways in response to allergen exposure. They correlated plasma levels of each of the biomarkers with lung function parameters (FEV1, FVC, and FEV1/FVC) in acute exacerbation and in remission. They found significant positive correlation between plasma 8-iso-PGF2α levels and the severity of airway obstruction detected by pulmonary function parameters. Also, Wood et al.  recently demonstrated the rise of lipid peroxidation in asthma measured by 8-iso-PGF2α concentration which was elevated 3–4 times in persistent asthmatics than the normal group. Judith and Mak  found that the plasma levels of 8-iso-PGF2α were significantly higher in patients with asthma in acute exacerbation and decreased in remission but remained elevated compared with healthy controls. One explanation could be its resistance to corticosteroids as demonstrated by Montuschi et al. , who reported that severe asthmatic patients treated with oral prednisolone had higher 8-iso-PGF2α levels in their exhaled breath condensates than mild and moderate asthma patients. They found increasing evidence that asthma is a disease associated with increased oxidative stress, with persistent elevated 8-iso-PGF2α levels during remission compared with healthy controls. Also, they found that FEV1, FVC, and FEV1/FVC were positively correlated with plasma 8-iso-PGF2α levels. Samitasa et al.  found increased levels of 8-iso-PGF2α in exhaled breath condensates in association with disease severity, in asthmatic adults and children. Keskin et al.  quantified that the F2-isoPGs level in BALF at baseline values was lower than its level 24 h after allergen instillation; moreover, exhaled 8-iso-PGF2α level in patients with moderate persistent asthma was higher than in the mild persistent group.
The present study showed that 8-iso-PGF2α levels in BALF and serum were higher in asthmatics than in healthy controls. Furthermore, their levels were increasing with asthma severity. These initial results appear to confirm the usefulness of determining these inflammatory mediators for the diagnosis and evaluation of severity of asthma, and also for investigating the relationship with oxidative stress and possible treatment control. However, further studies are needed to standardize the technique and validate the methods used to measure the mediators in order to define cutoff values for differentiating between healthy patients, asthmatics, and between different degrees of asthma severity.
In our study, we found a significant increase of TGFβ1 in BAL and serum in moderate and severe asthma compared with control, and in severe asthma compared with moderate asthma. Ozyilmaz et al.  measured plasma TGFβ1 level in three groups, 35 atopic, 35 nonatopic asthmatic patients, and 15 healthy control patients. Their levels were significantly higher in the asthmatic groups compared with the control, whereas they were similar among the atopic and nonatopic asthmatics. They found a positive correlation between uncontrolled asthma and plasma TGFβ1 level. They concluded that plasma TGFβ1 level may be a systemic marker of asthma control. Manuyakorn et al.  studied serum TGFβ1 levels in 31 atopic asthmatic patients and 34 nonatopic controls measured by enzyme-linked immunosorbent assay. Contrary to our results they found an elevation of serum TGFβ1 in the steroid-naive mild asthma group in comparison to the moderate asthma group, with no correlations between serum TGFβ1 levels and pulmonary function test parameters, and also duration of asthma or duration of inhaled corticosteroid treatment. Redington et al. , measured the levels of TGβ1 in BALF from clinically stable asthmatics and healthy control patients. Their levels were significantly higher in asthmatics than control patients. They conclude that basal TGFβ1 levels in the airways are elevated in asthma. Forno et al.  also observed elevated TGFβ1 expression in asthmatics. Joseph et al.  postulated that the mean value of the plasma TGFβ1 was significantly higher in nonatopic asthmatic patients compared with control individuals and atopic asthmatic patients. Hong et al.  reported that the serum expression of TGFβ1 in asthmatic children was lower than that of the control group. Jarrett et al.  conducted a study on 74 children with asthma attacks. They found that the TGFβ1 level in the asthmatic group increased significantly. In contrast to our results, Magnon et al.  reported decreased TGFβ1 expression in the asthmatic epithelium. Also, Lommatzsch Xiang and Qiu  reported no difference between the levels of serum TGFβ1 in atopic asthmatic adults and control patients. Moreover, Ceyhan et al.  concluded that the TGFβ1 level in serum had no predictive value in diagnostic use. Although our study confirmed the great role of TGFβ1 in asthma and assessed the variation in the level of TGFβ1 in BALF and serum of asthmatic patients, we did not have the information about histopathologicol changes in bronchial wall in response to TGFβ1.
Lastly, further studies need to be directed at the cellular, molecular, and genetic factors that are responsible for determining why only some people develop significant remodeling and why the type of remodeling that occurs can differ from patient to patient. The information that will come from these studies will profoundly affect our understanding of asthma pathogenesis, and will impact the types of strategies we use to combat this increasingly problematic disorder.
| Conclusion|| |
8-Iso-PGF2α and TGFβ1 levels in BAL and serum increase in moderate and severe asthmatic patients than in control normal patients. Their levels are correlated with the severity of asthma.
Further studies are needed to define cutoff values for differentiating between healthy patients and asthmatics and between different degrees of asthma severity.
Amgad Frahat contributed for concepts, design, definition of intellectual content, and manuscript review. Yousef Mansour contributed for definition of intellectual content and manuscript review. Ayman Eldib contributed for concepts, design, definition of intellectual content, literature search, clinical studies, experimental studies, data acquisition, data analysis, statistical analysis, manuscript preparation, manuscript editing, and manuscript review. Doaa Alsed contributed for clinical studies, experimental studies, statistical analysis, manuscript preparation, and manuscript editing.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Bousquet J, Mantzouranis E, Cruz AA, Khaled N, Baena-Cagnani CE, Bleecker ER et al.
Uniform definition of asthma severity, control, and exacerbations: document presented for theWorld Health Organization Consultation on Severe Asthma. J Allergy Clin Immunol
Pilacik B, Nofer T, Wasowicz W. F2
-Isoprostanes biomarkers of lipid peroxidation: their utility in evaluation of oxidative stress induced by toxic agents. Int J Occup Med Environ Health
Duvernelle C, Freund V, Frossard N. Transforming growth factor-beta and its role in asthma. Pulm Pharmacol Ther
Ghosh B, Sharma S, Nagpal K. Transforming growth factor beta1 gene (TGFβ1) prediction of susceptibility for immunological disease 2010
Howell JE, McAnulty RJ. TGF-beta: its role in asthma and therapeutic potential. Curr Drug Targets
Jingjing J, Gorge S. TGF-B2 reduces nitric oxide synthase mRNA through a ROCK-dependent pathway in airway epithelial cells. Curr Drug Targets
Mistry V, Doe C, Roach K, Morgan A, Wardlaw A, Pavord I et al.
Airway hyper responsiveness is dissociated from airway wall structural remodeling. J Allergy Clin Immunol
Moore W, Bleecker ER, Curran-Everett D, Erzurum SC, Ameredes BT, Bacharier L et al.
Characterization of the severe asthma phenotype by the National Heart, Lung and Blood Institute’s Severe Asthma Research Program. J Allergy Clin Immunol
Morrow JD, Roberts LJ. Isoprostanes: their role as an index of oxidant stress status in human pulmonary disease. Am J Respir Crit Care Med
Montuschi P, Corradi M, Ciabattoni G, Nightingale J, Kharitonov SA, Barnes PJ et al.
Increased 8-isoprostane, a marker of oxidative stress, in exhaled condensate of asthma patients. Am J Respir Crit Care Med
Wood LG, Fitzgerald DA, Gibson PG, Cooper DM, Garg ML. Lipid peroxidation as determined by plasma isoprostanes is related to disease severity in mild asthma. Lipids
Wood LC, Garg ML, Simpson JL, Mori TA, Croft KD, Wark PA, Gibson PG. Induced sputum 8-isoprostane concentrations in inflammatory airway diseases. Am J Respir Crit Care Med
Chang CH, Chang CH, Lui S. TGF1
is associated with asthma and modulates the severity of asthma. Resp Critical Care
Benayoun L, Druilhe A, Dombret MC, Aubier M, Pretolani M. Airway structural alterations selectively associated with sever asthma. Am J Respir Crit Care Med
Kharitonov SA, Barnes PJ. Increased 8-isoprostane, a marker of oxidative stress, in exhaled condensation in asthma patients. Am J Respir Crit Care Med
Dworski R, Murray J, Roberts J, Oates J, Morrow J, Fisher L et al.
Allergen- induced synthesis of F2
-isoprostanes in atopic asthmatics. Am J Respir Crit Care Med
Judith C, Mak W. Sustained elevation of systemic oxidative stress and inflammation in exacerbation and remission of asthma. Allergy
Samitasa K, Chorianopoulosb D, Vittorakisa S, Zervasa E, Economidoua E, Papatheodorouc G et al.
Exhaled cysteinyl-leukotrienes and 8-isoprostane in patients with asthma and their relation to clinical severity. Respir Med
Keskin O, Balaban S, Keskin M, Kucukosmanoglu E, Gogebakan B, Ozkars MY et al.
Relationship between exhalled leukottriene and 8 isoprostane level and asthma severity. Allergol Immunopathol (Madr)
Ozyilmaz E, Canbakan S, Capan N, Erturk A, Gulhan M. Correlation of plasma transforming growth factor beta1
with asthma control test. Asian Pac J Allergy Immunol
Manuyakorn W, Kamchaisatian W, Atamasirikul K, Sasisakulporn C, Direkwattanachai C, Benjaponpitak S. Serum TGF-beta1 in atopic asthma. Asian Pac J Allergy Immunol
Redington AE, Madden J, Frew AJ, Djukanovic R, Roche WR, Holgate ST, Howarth PH. Transforming growth factor-beta1
in asthma. Measurement in bronchoalveolar lavage fluid. Am J Respir Crit Care Med
Forno E, Lescher R, Strunk R, Weiss S, Fuhlbrigge A, Celedón JC. TGF-β1
expression in the asthmatics. J Allergy Clin Immunol
Joseph J, Benedict S, Badrinath P, Wassef S, Joseph M, Abdulkhalik S. Elevation of plasma transforming growth factor β1
levels in stable nonatopic asthma. Ann Allergy Asthma Immunol
Hong JG, Dong WF, Zhou XJ. Effect of montelukast sodium on TGF-β1 of peripheral blood mononuclear cells from children with mild persistent asthma. Eur J Cell Biol
Jarrett L, Chen JA, Figen J, Doğu L, Alişan N, Deniz E. Serum TGF-β, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase (TIMP-1) levels in childhood asthma. Turk J Med
Magnon L, Fukushima T, Yamasaki A, Harada T. TGF-β1 expression in the asthmatic epithelium. Am J Respir Cell Mol Biol
Lommatzsch Xiang S, Qiu D. Changes of CD4+CD25+ regulatory T cells, IL-10 and TGF-β1
levels in peripheral blood in asthma. Am J Respir Crit Care Med
Ceyhan N, Ronmark J, Platts-Mills R. TGF-β1
slevels in induced sputum samples of asthmatic subjects. Ann Allergy Asthma Immunol
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]