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Brief Glutamine Pretreatment Increases Alveolar Macrophage CD163/Heme Oxygenase-1/p38-MAPK Dephosphorylation Pathway and Decreases Capillary Damage but Not Neutrophil Recruitment in IL-1/LPS-Insufflated Rats.

Fernandez-Bustamante A, Agazio A, Wilson P, Elkins N, Domaleski L, He Q, Baer KA, Moss AF, Wischmeyer PE, Repine JE - PLoS ONE (2015)

Bottom Line: Glutamine (GLN) attenuates acute lung injury (ALI) but its effect on alveolar macrophages is unknown.GLN pretreatment before IL-1/LPS also significantly increased HO-1 concentrations and dephosphorylated p38-MAPK levels but not cytokine levels in alveolar macrophages.Short-term GLN pretreatment activates the anti-inflammatory CD163/HO-1/p38-MAPK dephosphorylation pathway of alveolar macrophages and decreases capillary damage but not neutrophil recruitment in IL-1/LPS-insufflated rats.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, University of Colorado SOM, Aurora, Colorado, United States of America; Webb-Waring Center, University of Colorado SOM, Aurora, Colorado, United States of America.

ABSTRACT

Background: Glutamine (GLN) attenuates acute lung injury (ALI) but its effect on alveolar macrophages is unknown. We hypothesized that GLN pretreatment would induce the anti-inflammatory CD163/heme oxygenase (HO)-1/p38-MAPK dephosphorylation pathway in alveolar macrophages and reduce ALI in rats insufflated with interleukin-1 (IL-1) and lipopolysaccharide (LPS).

Methods: Male Sprague-Dawley rats were randomized to the following groups: GLN-IL-1/LPS-, GLN+IL-1/LPS-, GLN-IL-1/LPS+, and GLN+IL-1/LPS+. GLN pretreatment was given via gavage (1 g/kg L-alanyl-L-glutamine) daily for 2 days. ALI was subsequently induced by insufflating 50 ng IL-1 followed by 5mg/kg E.coli LPS. After 24h, bronchoalveolar lavage (BAL) protein, lactate dehydrogenase (LDH) and neutrophil concentrations were analyzed. BAL alveolar macrophage CD163+ expression, HO-1 and p38-MAPK concentrations were measured, as well as alveolar macrophage tumor necrosis factor (TNF)-α and interleukin (IL)-10 concentrations. Histology and immunofluorescence studies were also performed.

Results: Following IL-1/LPS insufflation, GLN pretreated rats had significantly decreased BAL protein and LDH concentrations, but not BAL neutrophil counts, compared to non-GLN pretreated rats. The number of alveolar macrophages and the number of CD163+ macrophages were significantly increased in GLN pretreated IL-1/LPS-insufflated rats compared to non-GLN pretreated, IL-1/LPS-insufflated rats. GLN pretreatment before IL-1/LPS also significantly increased HO-1 concentrations and dephosphorylated p38-MAPK levels but not cytokine levels in alveolar macrophages. Immunofluorescence localized CD163 and HO-1 in alveolar macrophages.

Conclusion: Short-term GLN pretreatment activates the anti-inflammatory CD163/HO-1/p38-MAPK dephosphorylation pathway of alveolar macrophages and decreases capillary damage but not neutrophil recruitment in IL-1/LPS-insufflated rats.

No MeSH data available.


Related in: MedlinePlus

Measurements of HO-1 and dephosphorylated p38-MAPK concentrations in alveolar macrophages.The concentration of HO-1 in isolated alveolar macrophages (4a) was significantly increased exclusively by IL-1/LPS with no interaction or effect of GLN pretreatment. Fig 4b shows immunofluorescence representation of HO-1 stained lung regions from GLN-IL-1/LPS+ and GLN+IL-1/LPS+ groups. (white arrow points to zoomed cell in bottom right corner) (DAPI nuclear staining = blue; HO-1 = pink). Dephosphorylated p38-MAPK concentrations (4c) were significantly increased in the GLN+IL-1/LPS+ group compared to the other three groups, and both GLN and IL-1/LPS had a significantly interaction on this increase. (The significance of differences among the four groups was analyzed by two-way ANOVA. See statistical methods section for details on paired comparisons. Statistical significance was accepted as p<0.05: * compared to respective GLN- group, # compared to respective IL-1/LPS- group, ^ significant interaction between GLN and IL-1/LPS group).
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pone.0130764.g004: Measurements of HO-1 and dephosphorylated p38-MAPK concentrations in alveolar macrophages.The concentration of HO-1 in isolated alveolar macrophages (4a) was significantly increased exclusively by IL-1/LPS with no interaction or effect of GLN pretreatment. Fig 4b shows immunofluorescence representation of HO-1 stained lung regions from GLN-IL-1/LPS+ and GLN+IL-1/LPS+ groups. (white arrow points to zoomed cell in bottom right corner) (DAPI nuclear staining = blue; HO-1 = pink). Dephosphorylated p38-MAPK concentrations (4c) were significantly increased in the GLN+IL-1/LPS+ group compared to the other three groups, and both GLN and IL-1/LPS had a significantly interaction on this increase. (The significance of differences among the four groups was analyzed by two-way ANOVA. See statistical methods section for details on paired comparisons. Statistical significance was accepted as p<0.05: * compared to respective GLN- group, # compared to respective IL-1/LPS- group, ^ significant interaction between GLN and IL-1/LPS group).

Mentions: GLN and IL-1/LPS showed a significant interaction on macrophage HO-1 concentrations. Paired comparisons showed significantly higher macrophage HO-1 concentrations in GLN-pretreated IL-1/LPS-insufflated rats compared to both non-IL-1/LPS groups and the non-GLN IL-1/LPS-insufflated group (Fig 4a). Immunofluorescence located HO-1 staining in alveolar macrophages (Fig 4b).


Brief Glutamine Pretreatment Increases Alveolar Macrophage CD163/Heme Oxygenase-1/p38-MAPK Dephosphorylation Pathway and Decreases Capillary Damage but Not Neutrophil Recruitment in IL-1/LPS-Insufflated Rats.

Fernandez-Bustamante A, Agazio A, Wilson P, Elkins N, Domaleski L, He Q, Baer KA, Moss AF, Wischmeyer PE, Repine JE - PLoS ONE (2015)

Measurements of HO-1 and dephosphorylated p38-MAPK concentrations in alveolar macrophages.The concentration of HO-1 in isolated alveolar macrophages (4a) was significantly increased exclusively by IL-1/LPS with no interaction or effect of GLN pretreatment. Fig 4b shows immunofluorescence representation of HO-1 stained lung regions from GLN-IL-1/LPS+ and GLN+IL-1/LPS+ groups. (white arrow points to zoomed cell in bottom right corner) (DAPI nuclear staining = blue; HO-1 = pink). Dephosphorylated p38-MAPK concentrations (4c) were significantly increased in the GLN+IL-1/LPS+ group compared to the other three groups, and both GLN and IL-1/LPS had a significantly interaction on this increase. (The significance of differences among the four groups was analyzed by two-way ANOVA. See statistical methods section for details on paired comparisons. Statistical significance was accepted as p<0.05: * compared to respective GLN- group, # compared to respective IL-1/LPS- group, ^ significant interaction between GLN and IL-1/LPS group).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4493112&req=5

pone.0130764.g004: Measurements of HO-1 and dephosphorylated p38-MAPK concentrations in alveolar macrophages.The concentration of HO-1 in isolated alveolar macrophages (4a) was significantly increased exclusively by IL-1/LPS with no interaction or effect of GLN pretreatment. Fig 4b shows immunofluorescence representation of HO-1 stained lung regions from GLN-IL-1/LPS+ and GLN+IL-1/LPS+ groups. (white arrow points to zoomed cell in bottom right corner) (DAPI nuclear staining = blue; HO-1 = pink). Dephosphorylated p38-MAPK concentrations (4c) were significantly increased in the GLN+IL-1/LPS+ group compared to the other three groups, and both GLN and IL-1/LPS had a significantly interaction on this increase. (The significance of differences among the four groups was analyzed by two-way ANOVA. See statistical methods section for details on paired comparisons. Statistical significance was accepted as p<0.05: * compared to respective GLN- group, # compared to respective IL-1/LPS- group, ^ significant interaction between GLN and IL-1/LPS group).
Mentions: GLN and IL-1/LPS showed a significant interaction on macrophage HO-1 concentrations. Paired comparisons showed significantly higher macrophage HO-1 concentrations in GLN-pretreated IL-1/LPS-insufflated rats compared to both non-IL-1/LPS groups and the non-GLN IL-1/LPS-insufflated group (Fig 4a). Immunofluorescence located HO-1 staining in alveolar macrophages (Fig 4b).

Bottom Line: Glutamine (GLN) attenuates acute lung injury (ALI) but its effect on alveolar macrophages is unknown.GLN pretreatment before IL-1/LPS also significantly increased HO-1 concentrations and dephosphorylated p38-MAPK levels but not cytokine levels in alveolar macrophages.Short-term GLN pretreatment activates the anti-inflammatory CD163/HO-1/p38-MAPK dephosphorylation pathway of alveolar macrophages and decreases capillary damage but not neutrophil recruitment in IL-1/LPS-insufflated rats.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology, University of Colorado SOM, Aurora, Colorado, United States of America; Webb-Waring Center, University of Colorado SOM, Aurora, Colorado, United States of America.

ABSTRACT

Background: Glutamine (GLN) attenuates acute lung injury (ALI) but its effect on alveolar macrophages is unknown. We hypothesized that GLN pretreatment would induce the anti-inflammatory CD163/heme oxygenase (HO)-1/p38-MAPK dephosphorylation pathway in alveolar macrophages and reduce ALI in rats insufflated with interleukin-1 (IL-1) and lipopolysaccharide (LPS).

Methods: Male Sprague-Dawley rats were randomized to the following groups: GLN-IL-1/LPS-, GLN+IL-1/LPS-, GLN-IL-1/LPS+, and GLN+IL-1/LPS+. GLN pretreatment was given via gavage (1 g/kg L-alanyl-L-glutamine) daily for 2 days. ALI was subsequently induced by insufflating 50 ng IL-1 followed by 5mg/kg E.coli LPS. After 24h, bronchoalveolar lavage (BAL) protein, lactate dehydrogenase (LDH) and neutrophil concentrations were analyzed. BAL alveolar macrophage CD163+ expression, HO-1 and p38-MAPK concentrations were measured, as well as alveolar macrophage tumor necrosis factor (TNF)-α and interleukin (IL)-10 concentrations. Histology and immunofluorescence studies were also performed.

Results: Following IL-1/LPS insufflation, GLN pretreated rats had significantly decreased BAL protein and LDH concentrations, but not BAL neutrophil counts, compared to non-GLN pretreated rats. The number of alveolar macrophages and the number of CD163+ macrophages were significantly increased in GLN pretreated IL-1/LPS-insufflated rats compared to non-GLN pretreated, IL-1/LPS-insufflated rats. GLN pretreatment before IL-1/LPS also significantly increased HO-1 concentrations and dephosphorylated p38-MAPK levels but not cytokine levels in alveolar macrophages. Immunofluorescence localized CD163 and HO-1 in alveolar macrophages.

Conclusion: Short-term GLN pretreatment activates the anti-inflammatory CD163/HO-1/p38-MAPK dephosphorylation pathway of alveolar macrophages and decreases capillary damage but not neutrophil recruitment in IL-1/LPS-insufflated rats.

No MeSH data available.


Related in: MedlinePlus