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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 PGE2 and PGE3 synthesis and release. (a) COX-2 relative protein content in A549 cells, stimulated with BALF and treated with 50 μM 1 : 2 and 1 : 7 DHA/AA PUFA ratios. Data are expressed as “a.u.” (arbitrary units) of the densitometric values, normalized on the corresponding β-actin. The value of unstimulated cells was arbitrarily set as 100. Data are presented as mean ± standard deviation of 6 independent determinations (n = 6). The image is representative of all the WB experiments. (b) PGE2 (white bars) and PGE3 (black bars) content in culture media of A549 cells, stimulated with BALFs and treated with 50 μM 1 : 2 or 1 : 7 DHA/AA ratio. Data are presented as picograms (pg) of the indicated PG per mL. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; PG, prostaglandin; COX, cycloxygenase; BALF, bronchoalveolar lavage fluid; DHA, docosahexaenoic acid; AA, arachidonic acid; WB, western blot. ***P < 0.001 BALF versus unstimulated cells and 1 : 2 DHA/AA. **P < 0.01 1 : 7 DHA/AA versus all. *P < 0.05 1 : 2 DHA/AA versus unstimulated cells. ##P < 0.01 1 : 2 DHA/AA versus all.
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fig5: Effects of ω-3/ω-6 PUFA ratios on PGE2 and PGE3 synthesis and release. (a) COX-2 relative protein content in A549 cells, stimulated with BALF and treated with 50 μM 1 : 2 and 1 : 7 DHA/AA PUFA ratios. Data are expressed as “a.u.” (arbitrary units) of the densitometric values, normalized on the corresponding β-actin. The value of unstimulated cells was arbitrarily set as 100. Data are presented as mean ± standard deviation of 6 independent determinations (n = 6). The image is representative of all the WB experiments. (b) PGE2 (white bars) and PGE3 (black bars) content in culture media of A549 cells, stimulated with BALFs and treated with 50 μM 1 : 2 or 1 : 7 DHA/AA ratio. Data are presented as picograms (pg) of the indicated PG per mL. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; PG, prostaglandin; COX, cycloxygenase; BALF, bronchoalveolar lavage fluid; DHA, docosahexaenoic acid; AA, arachidonic acid; WB, western blot. ***P < 0.001 BALF versus unstimulated cells and 1 : 2 DHA/AA. **P < 0.01 1 : 7 DHA/AA versus all. *P < 0.05 1 : 2 DHA/AA versus unstimulated cells. ##P < 0.01 1 : 2 DHA/AA versus all.

Mentions: To verify whether in our experimental model there was a modulation of PG biosynthesis, we analyzed the content of COX-2. The results indicated that in response to BALF stimulus A549 cells increased by 2.5-fold the expression of COX-2 (Figure 5(a)) and produce a significant amount of PGE2 (Figure 5(b)). Noteworthy, even if COX-2 is the inducible form of COX, A549 cells express the enzyme constitutively [46]. According to Yang et al. [46], we found that this level of COX-2 expression was not associated with a PG synthesis (Figure 5(b)) in unstimulated A549.


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 PGE2 and PGE3 synthesis and release. (a) COX-2 relative protein content in A549 cells, stimulated with BALF and treated with 50 μM 1 : 2 and 1 : 7 DHA/AA PUFA ratios. Data are expressed as “a.u.” (arbitrary units) of the densitometric values, normalized on the corresponding β-actin. The value of unstimulated cells was arbitrarily set as 100. Data are presented as mean ± standard deviation of 6 independent determinations (n = 6). The image is representative of all the WB experiments. (b) PGE2 (white bars) and PGE3 (black bars) content in culture media of A549 cells, stimulated with BALFs and treated with 50 μM 1 : 2 or 1 : 7 DHA/AA ratio. Data are presented as picograms (pg) of the indicated PG per mL. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; PG, prostaglandin; COX, cycloxygenase; BALF, bronchoalveolar lavage fluid; DHA, docosahexaenoic acid; AA, arachidonic acid; WB, western blot. ***P < 0.001 BALF versus unstimulated cells and 1 : 2 DHA/AA. **P < 0.01 1 : 7 DHA/AA versus all. *P < 0.05 1 : 2 DHA/AA versus unstimulated cells. ##P < 0.01 1 : 2 DHA/AA versus all.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Effects of ω-3/ω-6 PUFA ratios on PGE2 and PGE3 synthesis and release. (a) COX-2 relative protein content in A549 cells, stimulated with BALF and treated with 50 μM 1 : 2 and 1 : 7 DHA/AA PUFA ratios. Data are expressed as “a.u.” (arbitrary units) of the densitometric values, normalized on the corresponding β-actin. The value of unstimulated cells was arbitrarily set as 100. Data are presented as mean ± standard deviation of 6 independent determinations (n = 6). The image is representative of all the WB experiments. (b) PGE2 (white bars) and PGE3 (black bars) content in culture media of A549 cells, stimulated with BALFs and treated with 50 μM 1 : 2 or 1 : 7 DHA/AA ratio. Data are presented as picograms (pg) of the indicated PG per mL. Data are presented as mean ± standard deviation of 12 independent determinations (n = 12). PUFA, polyunsaturated fatty acid; PG, prostaglandin; COX, cycloxygenase; BALF, bronchoalveolar lavage fluid; DHA, docosahexaenoic acid; AA, arachidonic acid; WB, western blot. ***P < 0.001 BALF versus unstimulated cells and 1 : 2 DHA/AA. **P < 0.01 1 : 7 DHA/AA versus all. *P < 0.05 1 : 2 DHA/AA versus unstimulated cells. ##P < 0.01 1 : 2 DHA/AA versus all.
Mentions: To verify whether in our experimental model there was a modulation of PG biosynthesis, we analyzed the content of COX-2. The results indicated that in response to BALF stimulus A549 cells increased by 2.5-fold the expression of COX-2 (Figure 5(a)) and produce a significant amount of PGE2 (Figure 5(b)). Noteworthy, even if COX-2 is the inducible form of COX, A549 cells express the enzyme constitutively [46]. According to Yang et al. [46], we found that this level of COX-2 expression was not associated with a PG synthesis (Figure 5(b)) in unstimulated A549.

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