Inhalation exposure to dihydroxyacetone promotes lung injury and pulmonary fibrosis in A/J mice

Figures

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  Fig. 1. Acute exposure to DHA induced inflammation in male mice. Male A/J mice were exposed to 5, 130, and 600 µg of DHA and total cell count per ml in BALF was measured (A) 1 h or (B) 24 h after exposure. Protein concentrations were also measured in these mice at (C) 1 h and (D) 24 h after exposure. A V-PLEX Proinflammatory cytokine panel was performed in plasma after acute exposures. Cytokine levels after exposure to 5, 130, and 600 µg of DHA for 1 h (E) and 24 h DHA exposure (F) are displayed using a heat map. KC/GRO levels were measured at the 5, 130, and 600 µg doses for 1 h DHA (G) and 24 h DHA (H). Mean values are displayed as mean ± SD of values. Statistical significance was determined by ANOVA with significance is displayed as follows: *p < 0.05, **p < 0.01, ****p < 0.0001.

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  Fig. 2. Acute exposure to DHA induced acute lung injury in male mice. Lung sections from male A/J mice were exposed to saline vehicle, 130, and 600 µg of DHA were examined 24 h after acute exposure. H&E staining was performed and imaged (A) to measure alveolar density (B) and acute lung injury (C). Scale bar is 200 µm. Mean ± SD of density or score are displayed. Statistical significance was determined by ANOVA with significance is displayed as follows: *p < 0.05, **p < 0.01, ****p < 0.0001.

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  Fig. 3. Sub-acute exposures to DHA promote inflammatory changes in male and female mice, and acute lung injury was observed in female mice. Male A/J mice were sub-acutely exposed to 5 µg of DHA for 2 weeks. BALF cell count (A) and protein levels (B) were measured in the vehicle control and DHA-exposed groups. Plasma from mice exposed sub-acutely to DHA was used to measure cytokine changes with the V-PLEX Proinflammatory panel. Cytokine levels in male A/J mice (C) and female A/J mice (D) are displayed by heat map with a separate KC/GRO graph for males (E) and females (F). Values are displayed as mean ± SD. Significance was determined by a Welch’s t-test and is displayed as follows: *p < 0.05.

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  Fig. 4. Sub-acute exposures to DHA promote acute lung injury in female mice. Female A/J mice were sub-acutely exposed to 5 µg of DHA for 2 weeks and H&E staining was performed on lungs (A). Alveolar density (B) and acute lung injury scores (C) were measured. Density and scores are displayed as mean ± SD. Significance was determined by a Welch’s t-test and is displayed as follows: **p < 0.01, ****p < 0.0001. Scale bar is 200 µm.

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  Fig. 5. Sub-acute exposures to DHA increase collagen deposition. Masson’s Tri-chrome staining was performed on female A/J mouse lungs sub-acutely exposed to 5 µg of DHA for 2 weeks, and collagen staining was quantified. The graphs are displayed as mean ± SD. Significance was determined by a Welch’s t-test and is displayed as follows: *p < 0.05. Scale bar is 200 µm.

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  Fig. 6. Lung function was altered after sub-acute exposures to DHA. The pressure-volume (PV) curve was measured using a FLEXIVENT machine. The graphs are displayed as mean ± SEM.