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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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Changes in the pulmonary resistance after challenge. Box plots of the pulmonary resistance (RL) within 10 min after challenge in asthmatic rats (n = 14, sensitized and challenged with OVA) and controls (n = 15, sensitized and challenged with normal saline). The RL values are expressed as differential values subtracted from the corresponding baseline values (Figure 1). Mild outliers (○) are cases with values between 1.5 and 3 box lengths from the upper or lower edge of the box. Extreme outliers (+) are cases with values more than 3 box lengths from the upper or lower edge of the box. As measured using the Kruskal-Wallis test, the differences in the RL at each time point in the asthmatic group, but not in the control group, were significant (p < 0.01). Significant differences in the RL were observed between the two groups at 1-6 min (* p < 0.05, ** p < 0.01, Mann-Whitney test) with a maximum increase in asthmatic rats observed 2 min after challenge.
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Figure 2: Changes in the pulmonary resistance after challenge. Box plots of the pulmonary resistance (RL) within 10 min after challenge in asthmatic rats (n = 14, sensitized and challenged with OVA) and controls (n = 15, sensitized and challenged with normal saline). The RL values are expressed as differential values subtracted from the corresponding baseline values (Figure 1). Mild outliers (○) are cases with values between 1.5 and 3 box lengths from the upper or lower edge of the box. Extreme outliers (+) are cases with values more than 3 box lengths from the upper or lower edge of the box. As measured using the Kruskal-Wallis test, the differences in the RL at each time point in the asthmatic group, but not in the control group, were significant (p < 0.01). Significant differences in the RL were observed between the two groups at 1-6 min (* p < 0.05, ** p < 0.01, Mann-Whitney test) with a maximum increase in asthmatic rats observed 2 min after challenge.

Mentions: The OVA challenge in sensitized rats induced a significant increase in the RL and, simultaneously, significant decreases in the Cdyn and RR compared with control rats. These alterations indicated an OVA-induced EAR. The most prominent change in asthmatic rats was in the RL with a maximum increase of 0.110 ± 0.115 kPa/ml/s (mean ± SD, p < 0.01, compared with the controls) 2 min after provocation (Figure 2). When compared with controls, the Cdyn showed a maximum decrease at 3 min, and the RR showed a maximum decrease at 2 min (data not shown).


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)

Changes in the pulmonary resistance after challenge. Box plots of the pulmonary resistance (RL) within 10 min after challenge in asthmatic rats (n = 14, sensitized and challenged with OVA) and controls (n = 15, sensitized and challenged with normal saline). The RL values are expressed as differential values subtracted from the corresponding baseline values (Figure 1). Mild outliers (○) are cases with values between 1.5 and 3 box lengths from the upper or lower edge of the box. Extreme outliers (+) are cases with values more than 3 box lengths from the upper or lower edge of the box. As measured using the Kruskal-Wallis test, the differences in the RL at each time point in the asthmatic group, but not in the control group, were significant (p < 0.01). Significant differences in the RL were observed between the two groups at 1-6 min (* p < 0.05, ** p < 0.01, Mann-Whitney test) with a maximum increase in asthmatic rats observed 2 min after challenge.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Changes in the pulmonary resistance after challenge. Box plots of the pulmonary resistance (RL) within 10 min after challenge in asthmatic rats (n = 14, sensitized and challenged with OVA) and controls (n = 15, sensitized and challenged with normal saline). The RL values are expressed as differential values subtracted from the corresponding baseline values (Figure 1). Mild outliers (○) are cases with values between 1.5 and 3 box lengths from the upper or lower edge of the box. Extreme outliers (+) are cases with values more than 3 box lengths from the upper or lower edge of the box. As measured using the Kruskal-Wallis test, the differences in the RL at each time point in the asthmatic group, but not in the control group, were significant (p < 0.01). Significant differences in the RL were observed between the two groups at 1-6 min (* p < 0.05, ** p < 0.01, Mann-Whitney test) with a maximum increase in asthmatic rats observed 2 min after challenge.
Mentions: The OVA challenge in sensitized rats induced a significant increase in the RL and, simultaneously, significant decreases in the Cdyn and RR compared with control rats. These alterations indicated an OVA-induced EAR. The most prominent change in asthmatic rats was in the RL with a maximum increase of 0.110 ± 0.115 kPa/ml/s (mean ± SD, p < 0.01, compared with the controls) 2 min after provocation (Figure 2). When compared with controls, the Cdyn showed a maximum decrease at 3 min, and the RR showed a maximum decrease at 2 min (data not shown).

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