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The early asthmatic response is associated with glycolysis, calcium binding and mitochondria activity as revealed by proteomic analysis in rats.

Xu YD, Cui JM, Wang Y, Yin LM, Gao CK, Liu YY, Yang YQ - Respir. Res. (2010)

Bottom Line: The inhalation of allergens by allergic asthmatics results in the early asthmatic response (EAR), which is characterized by acute airway obstruction beginning within a few minutes.Of these 44 protein spots, 42 corresponded to 36 unique proteins successfully identified using mass spectrometry.Using western blot and semi-quantitative RT-PCR, we confirmed the changes in expression of five selected proteins, which further supports our proteomic and bioinformatic analyses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Yue Yang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.

ABSTRACT

Background: The inhalation of allergens by allergic asthmatics results in the early asthmatic response (EAR), which is characterized by acute airway obstruction beginning within a few minutes. The EAR is the earliest indicator of the pathological progression of allergic asthma. Because the molecular mechanism underlying the EAR is not fully defined, this study will contribute to a better understanding of asthma.

Methods: In order to gain insight into the molecular basis of the EAR, we examined changes in protein expression patterns in the lung tissue of asthmatic rats during the EAR using 2-DE/MS-based proteomic techniques. Bioinformatic analysis of the proteomic data was then performed using PPI Spider and KEGG Spider to investigate the underlying molecular mechanism.

Results: In total, 44 differentially expressed protein spots were detected in the 2-DE gels. Of these 44 protein spots, 42 corresponded to 36 unique proteins successfully identified using mass spectrometry. During subsequent bioinformatic analysis, the gene ontology classification, the protein-protein interaction networking and the biological pathway exploration demonstrated that the identified proteins were mainly involved in glycolysis, calcium binding and mitochondrial activity. Using western blot and semi-quantitative RT-PCR, we confirmed the changes in expression of five selected proteins, which further supports our proteomic and bioinformatic analyses.

Conclusions: Our results reveal that the allergen-induced EAR in asthmatic rats is associated with glycolysis, calcium binding and mitochondrial activity, which could establish a functional network in which calcium binding may play a central role in promoting the progression of asthma.

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Validation of the 2-DE proteomic data by western blot analysis. (A) A representative western blot visualizing the ERp29, RhoGDI2 and S100A8 expression levels. β-actin was used to demonstrate equal loading. NC = normal control group (n = 15) and AS = asthmatic group (n = 14). (B) The densitometric quantification of individual protein is expressed as the fold change compared to β-actin. The data are presented as the mean ± SD of triplicate experiments, and similar results were seen in all experiments. Statistical comparisons were made using Student's t-test. *p < 0.05, **p < 0.01 comparing the asthmatic group with the control group. The expression pattern of these proteins is in agreement with the 2-DE results.
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Figure 6: Validation of the 2-DE proteomic data by western blot analysis. (A) A representative western blot visualizing the ERp29, RhoGDI2 and S100A8 expression levels. β-actin was used to demonstrate equal loading. NC = normal control group (n = 15) and AS = asthmatic group (n = 14). (B) The densitometric quantification of individual protein is expressed as the fold change compared to β-actin. The data are presented as the mean ± SD of triplicate experiments, and similar results were seen in all experiments. Statistical comparisons were made using Student's t-test. *p < 0.05, **p < 0.01 comparing the asthmatic group with the control group. The expression pattern of these proteins is in agreement with the 2-DE results.

Mentions: To verify the 2-DE findings, we further examined the expression of RhoGDI2, ERp29 and S100A8 using western blots. Compared with the controls, the expression levels of RhoGDI2 and S100A8 were significantly increased (p < 0.01 and p < 0.05, respectively) in the asthmatic rats, whereas the expression of ERp29 was significantly decreased (p < 0.01) in the asthmatic rats (Figure 6). These expression patterns were consistent with the patterns detected in the 2-DE analysis. These results confirm the reliability of this comparative proteomic study.


The early asthmatic response is associated with glycolysis, calcium binding and mitochondria activity as revealed by proteomic analysis in rats.

Xu YD, Cui JM, Wang Y, Yin LM, Gao CK, Liu YY, Yang YQ - Respir. Res. (2010)

Validation of the 2-DE proteomic data by western blot analysis. (A) A representative western blot visualizing the ERp29, RhoGDI2 and S100A8 expression levels. β-actin was used to demonstrate equal loading. NC = normal control group (n = 15) and AS = asthmatic group (n = 14). (B) The densitometric quantification of individual protein is expressed as the fold change compared to β-actin. The data are presented as the mean ± SD of triplicate experiments, and similar results were seen in all experiments. Statistical comparisons were made using Student's t-test. *p < 0.05, **p < 0.01 comparing the asthmatic group with the control group. The expression pattern of these proteins is in agreement with the 2-DE results.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2925830&req=5

Figure 6: Validation of the 2-DE proteomic data by western blot analysis. (A) A representative western blot visualizing the ERp29, RhoGDI2 and S100A8 expression levels. β-actin was used to demonstrate equal loading. NC = normal control group (n = 15) and AS = asthmatic group (n = 14). (B) The densitometric quantification of individual protein is expressed as the fold change compared to β-actin. The data are presented as the mean ± SD of triplicate experiments, and similar results were seen in all experiments. Statistical comparisons were made using Student's t-test. *p < 0.05, **p < 0.01 comparing the asthmatic group with the control group. The expression pattern of these proteins is in agreement with the 2-DE results.
Mentions: To verify the 2-DE findings, we further examined the expression of RhoGDI2, ERp29 and S100A8 using western blots. Compared with the controls, the expression levels of RhoGDI2 and S100A8 were significantly increased (p < 0.01 and p < 0.05, respectively) in the asthmatic rats, whereas the expression of ERp29 was significantly decreased (p < 0.01) in the asthmatic rats (Figure 6). These expression patterns were consistent with the patterns detected in the 2-DE analysis. These results confirm the reliability of this comparative proteomic study.

Bottom Line: The inhalation of allergens by allergic asthmatics results in the early asthmatic response (EAR), which is characterized by acute airway obstruction beginning within a few minutes.Of these 44 protein spots, 42 corresponded to 36 unique proteins successfully identified using mass spectrometry.Using western blot and semi-quantitative RT-PCR, we confirmed the changes in expression of five selected proteins, which further supports our proteomic and bioinformatic analyses.

View Article: PubMed Central - HTML - PubMed

Affiliation: Yue Yang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.

ABSTRACT

Background: The inhalation of allergens by allergic asthmatics results in the early asthmatic response (EAR), which is characterized by acute airway obstruction beginning within a few minutes. The EAR is the earliest indicator of the pathological progression of allergic asthma. Because the molecular mechanism underlying the EAR is not fully defined, this study will contribute to a better understanding of asthma.

Methods: In order to gain insight into the molecular basis of the EAR, we examined changes in protein expression patterns in the lung tissue of asthmatic rats during the EAR using 2-DE/MS-based proteomic techniques. Bioinformatic analysis of the proteomic data was then performed using PPI Spider and KEGG Spider to investigate the underlying molecular mechanism.

Results: In total, 44 differentially expressed protein spots were detected in the 2-DE gels. Of these 44 protein spots, 42 corresponded to 36 unique proteins successfully identified using mass spectrometry. During subsequent bioinformatic analysis, the gene ontology classification, the protein-protein interaction networking and the biological pathway exploration demonstrated that the identified proteins were mainly involved in glycolysis, calcium binding and mitochondrial activity. Using western blot and semi-quantitative RT-PCR, we confirmed the changes in expression of five selected proteins, which further supports our proteomic and bioinformatic analyses.

Conclusions: Our results reveal that the allergen-induced EAR in asthmatic rats is associated with glycolysis, calcium binding and mitochondrial activity, which could establish a functional network in which calcium binding may play a central role in promoting the progression of asthma.

Show MeSH
Related in: MedlinePlus