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The impact of surfactant protein-A on ozone-induced changes in the mouse bronchoalveolar lavage proteome.

Haque R, Umstead TM, Freeman WM, Floros J, Phelps DS - Proteome Sci (2009)

Bottom Line: We identified 66 proteins and focused our analysis on these proteins.Many of them fell into three categories: defense and immunity; redox regulation; and protein metabolism, modification and chaperones.Most of the proteins in the redox group were decreased, the proteins involved in protein metabolism increased, and roughly equal numbers of increases and decreases were seen in the defense and immunity group.

View Article: PubMed Central - HTML - PubMed

Affiliation: Penn State Center for Host defense, Inflammation, and Lung Disease Research and the Department of Pediatrics, Penn State College of Medicine, Hershey, USA. rhaque1@hmc.psu.edu

ABSTRACT

Background: Ozone is a major component of air pollution. Exposure to this powerful oxidizing agent can cause or exacerbate many lung conditions, especially those involving innate immunity. Surfactant protein-A (SP-A) plays many roles in innate immunity by participating directly in host defense as it exerts opsonin function, or indirectly via its ability to regulate alveolar macrophages and other innate immune cells. The mechanism(s) responsible for ozone-induced pathophysiology, while likely related to oxidative stress, are not well understood.

Methods: We employed 2-dimensional difference gel electrophoresis (2D-DIGE), a discovery proteomics approach, coupled with MALDI-ToF/ToF to compare the bronchoalveolar lavage (BAL) proteomes in wild type (WT) and SP-A knockout (KO) mice and to assess the impact of ozone or filtered air on the expression of BAL proteins. Using the PANTHER database and the published literature most identified proteins were placed into three functional groups.

Results: We identified 66 proteins and focused our analysis on these proteins. Many of them fell into three categories: defense and immunity; redox regulation; and protein metabolism, modification and chaperones. In response to the oxidative stress of acute ozone exposure (2 ppm; 3 hours) there were many significant changes in levels of expression of proteins in these groups. Most of the proteins in the redox group were decreased, the proteins involved in protein metabolism increased, and roughly equal numbers of increases and decreases were seen in the defense and immunity group. Responses between WT and KO mice were similar in many respects. However, the percent change was consistently greater in the KO mice and there were more changes that achieved statistical significance in the KO mice, with levels of expression in filtered air-exposed KO mice being closer to ozone-exposed WT mice than to filtered air-exposed WT mice.

Conclusion: We postulate that SP-A plays a role in reactive oxidant scavenging in WT mice and that its absence in the KO mice in the presence or absence of ozone exposure results in more pronounced, and presumably chronic, oxidative stress.

No MeSH data available.


Related in: MedlinePlus

Principal component analysis (PCA) of 2D-DIGE data. This figure depicts a principal component analysis in which principal components (PC) 1 and 2 are plotted for all 454 matched protein spots. PC1 accounted for 42.54% of the variance and PC2 accounted for 12.76% and segregated the groups by ozone exposure and SP-A deletion, respectively. Circles represent the WT mice and triangles represent KO mice. The open symbols represent samples that were exposed to FA and the black symbols represent those exposed to ozone.
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Figure 3: Principal component analysis (PCA) of 2D-DIGE data. This figure depicts a principal component analysis in which principal components (PC) 1 and 2 are plotted for all 454 matched protein spots. PC1 accounted for 42.54% of the variance and PC2 accounted for 12.76% and segregated the groups by ozone exposure and SP-A deletion, respectively. Circles represent the WT mice and triangles represent KO mice. The open symbols represent samples that were exposed to FA and the black symbols represent those exposed to ozone.

Mentions: To examine the relationship of the samples in each group as well as the groups to each other based on the proteomic profile, principal components analysis of the data (Figure 3) was performed using all 454 matched protein spots. Principal components 1 and 2 accounted for 42.54% and 12.76% of the study variance, respectively. Principal component 1 segregated the samples by ozone exposure and principal component 2 by strain. The contributions to the relative variance of the two principal components (42.54% vs 12.76%) indicate that the effect of ozone exposure (principal component 1) on changes in the BAL proteome is greater than that of SP-A deletion (principal component 2). Each of the 16 independent animals (4/group) represented by the markers in the figure represents the combined weighted average of the first two principal components for each of the 454 protein spots. Each of the four experimental groups are tightly clustered with no overlap among groups.


The impact of surfactant protein-A on ozone-induced changes in the mouse bronchoalveolar lavage proteome.

Haque R, Umstead TM, Freeman WM, Floros J, Phelps DS - Proteome Sci (2009)

Principal component analysis (PCA) of 2D-DIGE data. This figure depicts a principal component analysis in which principal components (PC) 1 and 2 are plotted for all 454 matched protein spots. PC1 accounted for 42.54% of the variance and PC2 accounted for 12.76% and segregated the groups by ozone exposure and SP-A deletion, respectively. Circles represent the WT mice and triangles represent KO mice. The open symbols represent samples that were exposed to FA and the black symbols represent those exposed to ozone.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2666657&req=5

Figure 3: Principal component analysis (PCA) of 2D-DIGE data. This figure depicts a principal component analysis in which principal components (PC) 1 and 2 are plotted for all 454 matched protein spots. PC1 accounted for 42.54% of the variance and PC2 accounted for 12.76% and segregated the groups by ozone exposure and SP-A deletion, respectively. Circles represent the WT mice and triangles represent KO mice. The open symbols represent samples that were exposed to FA and the black symbols represent those exposed to ozone.
Mentions: To examine the relationship of the samples in each group as well as the groups to each other based on the proteomic profile, principal components analysis of the data (Figure 3) was performed using all 454 matched protein spots. Principal components 1 and 2 accounted for 42.54% and 12.76% of the study variance, respectively. Principal component 1 segregated the samples by ozone exposure and principal component 2 by strain. The contributions to the relative variance of the two principal components (42.54% vs 12.76%) indicate that the effect of ozone exposure (principal component 1) on changes in the BAL proteome is greater than that of SP-A deletion (principal component 2). Each of the 16 independent animals (4/group) represented by the markers in the figure represents the combined weighted average of the first two principal components for each of the 454 protein spots. Each of the four experimental groups are tightly clustered with no overlap among groups.

Bottom Line: We identified 66 proteins and focused our analysis on these proteins.Many of them fell into three categories: defense and immunity; redox regulation; and protein metabolism, modification and chaperones.Most of the proteins in the redox group were decreased, the proteins involved in protein metabolism increased, and roughly equal numbers of increases and decreases were seen in the defense and immunity group.

View Article: PubMed Central - HTML - PubMed

Affiliation: Penn State Center for Host defense, Inflammation, and Lung Disease Research and the Department of Pediatrics, Penn State College of Medicine, Hershey, USA. rhaque1@hmc.psu.edu

ABSTRACT

Background: Ozone is a major component of air pollution. Exposure to this powerful oxidizing agent can cause or exacerbate many lung conditions, especially those involving innate immunity. Surfactant protein-A (SP-A) plays many roles in innate immunity by participating directly in host defense as it exerts opsonin function, or indirectly via its ability to regulate alveolar macrophages and other innate immune cells. The mechanism(s) responsible for ozone-induced pathophysiology, while likely related to oxidative stress, are not well understood.

Methods: We employed 2-dimensional difference gel electrophoresis (2D-DIGE), a discovery proteomics approach, coupled with MALDI-ToF/ToF to compare the bronchoalveolar lavage (BAL) proteomes in wild type (WT) and SP-A knockout (KO) mice and to assess the impact of ozone or filtered air on the expression of BAL proteins. Using the PANTHER database and the published literature most identified proteins were placed into three functional groups.

Results: We identified 66 proteins and focused our analysis on these proteins. Many of them fell into three categories: defense and immunity; redox regulation; and protein metabolism, modification and chaperones. In response to the oxidative stress of acute ozone exposure (2 ppm; 3 hours) there were many significant changes in levels of expression of proteins in these groups. Most of the proteins in the redox group were decreased, the proteins involved in protein metabolism increased, and roughly equal numbers of increases and decreases were seen in the defense and immunity group. Responses between WT and KO mice were similar in many respects. However, the percent change was consistently greater in the KO mice and there were more changes that achieved statistical significance in the KO mice, with levels of expression in filtered air-exposed KO mice being closer to ozone-exposed WT mice than to filtered air-exposed WT mice.

Conclusion: We postulate that SP-A plays a role in reactive oxidant scavenging in WT mice and that its absence in the KO mice in the presence or absence of ozone exposure results in more pronounced, and presumably chronic, oxidative stress.

No MeSH data available.


Related in: MedlinePlus