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The Omega-3 Fatty Acid Docosahexaenoic Acid Modulates Inflammatory Mediator Release in Human Alveolar Cells Exposed to Bronchoalveolar Lavage Fluid of ARDS Patients.

Cotogni P, Trombetta A, Muzio G, Maggiora M, Canuto RA - Biomed Res Int (2015)

Bottom Line: The proinflammatory cytokine release was reduced by the 1 : 2 ratio (P < 0.01 to <0.001) but was increased by the 1 : 7 ratio (P < 0.01).The 1 : 2 ratio reduced COX-2 and PGE2 (P < 0.001) as well as NF-κB translocation into the nucleus (P < 0.01), while it increased activation of PPARγ and IL-10 release (P < 0.001).Conclusion.

View Article: PubMed Central - PubMed

Affiliation: Anesthesiology and Intensive Care, Department of Medicine, S. Giovanni Battista Hospital, University of Turin, Via A.M. Dogliotti 14, 10126 Turin, Italy.

ABSTRACT

Background: This study investigated whether the 1 : 2 ω-3/ω-6 ratio may reduce proinflammatory response in human alveolar cells (A549) exposed to an ex vivo inflammatory stimulus (bronchoalveolar lavage fluid (BALF) of acute respiratory distress syndrome (ARDS) patients). Methods. We exposed A549 cells to the BALF collected from 12 ARDS patients. After 18 hours, fatty acids (FA) were added as docosahexaenoic acid (DHA, ω-3) and arachidonic acid (AA, ω-6) in two ratios (1 : 2 or 1 : 7). 24 hours later, in culture supernatants were evaluated cytokines (TNF-α, IL-6, IL-8, and IL-10) and prostaglandins (PGE2 and PGE3) release. The FA percentage content in A549 membrane phospholipids, content of COX-2, level of PPARγ, and NF-κB binding activity were determined.

Results: The 1 : 2 DHA/AA ratio reversed the baseline predominance of ω-6 over ω-3 in the cell membranes (P < 0.001). The proinflammatory cytokine release was reduced by the 1 : 2 ratio (P < 0.01 to <0.001) but was increased by the 1 : 7 ratio (P < 0.01). The 1 : 2 ratio reduced COX-2 and PGE2 (P < 0.001) as well as NF-κB translocation into the nucleus (P < 0.01), while it increased activation of PPARγ and IL-10 release (P < 0.001). Conclusion. This study demonstrated that shifting the FA supply from ω-6 to ω-3 decreased proinflammatory mediator release in human alveolar cells exposed to BALF of ARDS patients.

No MeSH data available.


Related in: MedlinePlus

Effects of ω-3/ω-6 PUFA ratios on BALF induced cytokine release from A549 cells. (a) TNF-α proinflammatory cytokine release from A549 cells, stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (b) IL-6 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA (ω-3/ω-6) PUFA ratios. (c) IL-8 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (d) IL-10 anti-inflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. In each panel, data are presented as picograms (pg) of the indicated cytokine per million cells. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; BALF, bronchoalveolar lavage fluid; TNF, tumor necrosis factor; DHA, docosahexaenoic acid; AA, arachidonic acid; IL, interleukin; LPS, lipopolysaccharide. ***P < 0.001 1 : 2 DHA/AA versus all. **P < 0.01 1 : 2 DHA/AA versus all. ##P < 0.01 1 : 7 DHA/AA versus LPS and BALF.
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fig2: Effects of ω-3/ω-6 PUFA ratios on BALF induced cytokine release from A549 cells. (a) TNF-α proinflammatory cytokine release from A549 cells, stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (b) IL-6 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA (ω-3/ω-6) PUFA ratios. (c) IL-8 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (d) IL-10 anti-inflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. In each panel, data are presented as picograms (pg) of the indicated cytokine per million cells. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; BALF, bronchoalveolar lavage fluid; TNF, tumor necrosis factor; DHA, docosahexaenoic acid; AA, arachidonic acid; IL, interleukin; LPS, lipopolysaccharide. ***P < 0.001 1 : 2 DHA/AA versus all. **P < 0.01 1 : 2 DHA/AA versus all. ##P < 0.01 1 : 7 DHA/AA versus LPS and BALF.

Mentions: In a previous study, we demonstrated that human alveolar cells (A549) release proinflammatory cytokines (TNF-α, IL-6, and IL-8) in the culture medium when challenged with a proinflammatory stimulus such as LPS, suggesting that the alveolar epithelium has a role in the hyperinflammatory response associated with ARDS [32]. Since LPS is an artificial stimulus, in this study we challenged A549 with BALF of ARDS patients. In this study, BALF stimulation induced a proinflammatory response from A549 cells as demonstrated by release of inflammatory cytokines: TNF-α (Figure 2(a)), IL-6 (Figure 2(b)), and IL-8 (Figure 2(c)) while BALF did not elicit anti-inflammatory IL-10 release (Figure 2(d)). These results demonstrated that the pattern of cytokine release of A549 cells exposed to BALF is similar to that elicited by LPS in the previous study.


The Omega-3 Fatty Acid Docosahexaenoic Acid Modulates Inflammatory Mediator Release in Human Alveolar Cells Exposed to Bronchoalveolar Lavage Fluid of ARDS Patients.

Cotogni P, Trombetta A, Muzio G, Maggiora M, Canuto RA - Biomed Res Int (2015)

Effects of ω-3/ω-6 PUFA ratios on BALF induced cytokine release from A549 cells. (a) TNF-α proinflammatory cytokine release from A549 cells, stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (b) IL-6 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA (ω-3/ω-6) PUFA ratios. (c) IL-8 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (d) IL-10 anti-inflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. In each panel, data are presented as picograms (pg) of the indicated cytokine per million cells. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; BALF, bronchoalveolar lavage fluid; TNF, tumor necrosis factor; DHA, docosahexaenoic acid; AA, arachidonic acid; IL, interleukin; LPS, lipopolysaccharide. ***P < 0.001 1 : 2 DHA/AA versus all. **P < 0.01 1 : 2 DHA/AA versus all. ##P < 0.01 1 : 7 DHA/AA versus LPS and BALF.
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Related In: Results  -  Collection

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fig2: Effects of ω-3/ω-6 PUFA ratios on BALF induced cytokine release from A549 cells. (a) TNF-α proinflammatory cytokine release from A549 cells, stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (b) IL-6 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA (ω-3/ω-6) PUFA ratios. (c) IL-8 proinflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. (d) IL-10 anti-inflammatory cytokine release from A549 cells stimulated with BALF and treated with 1 : 2 and 1 : 7 DHA/AA PUFA ratios. In each panel, data are presented as picograms (pg) of the indicated cytokine per million cells. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; BALF, bronchoalveolar lavage fluid; TNF, tumor necrosis factor; DHA, docosahexaenoic acid; AA, arachidonic acid; IL, interleukin; LPS, lipopolysaccharide. ***P < 0.001 1 : 2 DHA/AA versus all. **P < 0.01 1 : 2 DHA/AA versus all. ##P < 0.01 1 : 7 DHA/AA versus LPS and BALF.
Mentions: In a previous study, we demonstrated that human alveolar cells (A549) release proinflammatory cytokines (TNF-α, IL-6, and IL-8) in the culture medium when challenged with a proinflammatory stimulus such as LPS, suggesting that the alveolar epithelium has a role in the hyperinflammatory response associated with ARDS [32]. Since LPS is an artificial stimulus, in this study we challenged A549 with BALF of ARDS patients. In this study, BALF stimulation induced a proinflammatory response from A549 cells as demonstrated by release of inflammatory cytokines: TNF-α (Figure 2(a)), IL-6 (Figure 2(b)), and IL-8 (Figure 2(c)) while BALF did not elicit anti-inflammatory IL-10 release (Figure 2(d)). These results demonstrated that the pattern of cytokine release of A549 cells exposed to BALF is similar to that elicited by LPS in the previous study.

Bottom Line: The proinflammatory cytokine release was reduced by the 1 : 2 ratio (P < 0.01 to <0.001) but was increased by the 1 : 7 ratio (P < 0.01).The 1 : 2 ratio reduced COX-2 and PGE2 (P < 0.001) as well as NF-κB translocation into the nucleus (P < 0.01), while it increased activation of PPARγ and IL-10 release (P < 0.001).Conclusion.

View Article: PubMed Central - PubMed

Affiliation: Anesthesiology and Intensive Care, Department of Medicine, S. Giovanni Battista Hospital, University of Turin, Via A.M. Dogliotti 14, 10126 Turin, Italy.

ABSTRACT

Background: This study investigated whether the 1 : 2 ω-3/ω-6 ratio may reduce proinflammatory response in human alveolar cells (A549) exposed to an ex vivo inflammatory stimulus (bronchoalveolar lavage fluid (BALF) of acute respiratory distress syndrome (ARDS) patients). Methods. We exposed A549 cells to the BALF collected from 12 ARDS patients. After 18 hours, fatty acids (FA) were added as docosahexaenoic acid (DHA, ω-3) and arachidonic acid (AA, ω-6) in two ratios (1 : 2 or 1 : 7). 24 hours later, in culture supernatants were evaluated cytokines (TNF-α, IL-6, IL-8, and IL-10) and prostaglandins (PGE2 and PGE3) release. The FA percentage content in A549 membrane phospholipids, content of COX-2, level of PPARγ, and NF-κB binding activity were determined.

Results: The 1 : 2 DHA/AA ratio reversed the baseline predominance of ω-6 over ω-3 in the cell membranes (P < 0.001). The proinflammatory cytokine release was reduced by the 1 : 2 ratio (P < 0.01 to <0.001) but was increased by the 1 : 7 ratio (P < 0.01). The 1 : 2 ratio reduced COX-2 and PGE2 (P < 0.001) as well as NF-κB translocation into the nucleus (P < 0.01), while it increased activation of PPARγ and IL-10 release (P < 0.001). Conclusion. This study demonstrated that shifting the FA supply from ω-6 to ω-3 decreased proinflammatory mediator release in human alveolar cells exposed to BALF of ARDS patients.

No MeSH data available.


Related in: MedlinePlus