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Resolvins: a family of bioactive products of omega-3 fatty acid transformation circuits initiated by aspirin treatment that counter proinflammation signals.

Serhan CN, Hong S, Gronert K, Colgan SP, Devchand PR, Mirick G, Moussignac RL - J. Exp. Med. (2002)

Bottom Line: Human neutrophils transformed COX-2-ASA-derived 17R-hydroxy-DHA into two sets of novel di- and trihydroxy products; one initiated via oxygenation at carbon 7 and the other at carbon 4.These compounds inhibited (IC(50) approximately 50 pM) microglial cell cytokine expression and in vivo dermal inflammation and peritonitis at ng doses, reducing 40-80% leukocytic exudates.These results indicate that exudates, vascular, leukocytes and neural cells treated with aspirin convert DHA to novel 17R-hydroxy series of docosanoids that are potent regulators.

View Article: PubMed Central - PubMed

Affiliation: Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesiology, Perioperative, and Pain Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA. cnserhan@zeus.bwh.harvard.edu

ABSTRACT
Aspirin (ASA) is unique among current therapies because it acetylates cyclooxygenase (COX)-2 enabling the biosynthesis of R-containing precursors of endogenous antiinflammatory mediators. Here, we report that lipidomic analysis of exudates obtained in the resolution phase from mice treated with ASA and docosahexaenoic acid (DHA) (C22:6) produce a novel family of bioactive 17R-hydroxy-containing di- and tri-hydroxy-docosanoids termed resolvins. Murine brain treated with aspirin produced endogenous 17R-hydroxydocosahexaenoic acid as did human microglial cells. Human COX-2 converted DHA to 13-hydroxy-DHA that switched with ASA to 17R-HDHA that also proved a major route in hypoxic endothelial cells. Human neutrophils transformed COX-2-ASA-derived 17R-hydroxy-DHA into two sets of novel di- and trihydroxy products; one initiated via oxygenation at carbon 7 and the other at carbon 4. These compounds inhibited (IC(50) approximately 50 pM) microglial cell cytokine expression and in vivo dermal inflammation and peritonitis at ng doses, reducing 40-80% leukocytic exudates. These results indicate that exudates, vascular, leukocytes and neural cells treated with aspirin convert DHA to novel 17R-hydroxy series of docosanoids that are potent regulators. These biosynthetic pathways utilize omega-3 DHA and EPA during multicellular events in resolution to produce a family of protective compounds, i.e., resolvins, that enhance proresolution status.

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Related in: MedlinePlus

Novel ASA triggered HDHA products generated by human recombinant COX-2 ASA. 17R-HDHA. Human recombinant COX-2 treated in the presence and absence of 2 mM ASA was incubated with DHA (10 μM, 30 min, 37°C). Incubations were stopped with 2 ml cold methanol, extracted and taken for LC-MS-MS analyses. Results are representative of incubations from more than eight separate experiments, some with 1-14C-labeled DHA. (Top) LC-MS-MS chromatogram of m/z 343 showing the presence of mono-HDHA. (Bottom) MS-MS spectrum of (left) 13-HDHA without ASA treatment and (right) 17R-HDHA with ASA treatment.
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fig2: Novel ASA triggered HDHA products generated by human recombinant COX-2 ASA. 17R-HDHA. Human recombinant COX-2 treated in the presence and absence of 2 mM ASA was incubated with DHA (10 μM, 30 min, 37°C). Incubations were stopped with 2 ml cold methanol, extracted and taken for LC-MS-MS analyses. Results are representative of incubations from more than eight separate experiments, some with 1-14C-labeled DHA. (Top) LC-MS-MS chromatogram of m/z 343 showing the presence of mono-HDHA. (Bottom) MS-MS spectrum of (left) 13-HDHA without ASA treatment and (right) 17R-HDHA with ASA treatment.

Mentions: Human recombinant COX-2 was overexpressed in Sf9 insect cells (American Type Culture Collection). The microsomal fractions (∼8 μl) were suspended in Tris (100 mM, pH 8.0) as in reference 35. ASA was incubated (∼2 mM, 37°C, 30 min) with COX-2 before addition of DHA (10 μM), or in some experiments [1-14C]-labeled DHA (American Radiolabeled Chemicals, Inc.), and conversions were monitored in parallel using [1-14C]-labeled C20:4 (with ASA) (as in Fig. 2 ; also see text).


Resolvins: a family of bioactive products of omega-3 fatty acid transformation circuits initiated by aspirin treatment that counter proinflammation signals.

Serhan CN, Hong S, Gronert K, Colgan SP, Devchand PR, Mirick G, Moussignac RL - J. Exp. Med. (2002)

Novel ASA triggered HDHA products generated by human recombinant COX-2 ASA. 17R-HDHA. Human recombinant COX-2 treated in the presence and absence of 2 mM ASA was incubated with DHA (10 μM, 30 min, 37°C). Incubations were stopped with 2 ml cold methanol, extracted and taken for LC-MS-MS analyses. Results are representative of incubations from more than eight separate experiments, some with 1-14C-labeled DHA. (Top) LC-MS-MS chromatogram of m/z 343 showing the presence of mono-HDHA. (Bottom) MS-MS spectrum of (left) 13-HDHA without ASA treatment and (right) 17R-HDHA with ASA treatment.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2194036&req=5

fig2: Novel ASA triggered HDHA products generated by human recombinant COX-2 ASA. 17R-HDHA. Human recombinant COX-2 treated in the presence and absence of 2 mM ASA was incubated with DHA (10 μM, 30 min, 37°C). Incubations were stopped with 2 ml cold methanol, extracted and taken for LC-MS-MS analyses. Results are representative of incubations from more than eight separate experiments, some with 1-14C-labeled DHA. (Top) LC-MS-MS chromatogram of m/z 343 showing the presence of mono-HDHA. (Bottom) MS-MS spectrum of (left) 13-HDHA without ASA treatment and (right) 17R-HDHA with ASA treatment.
Mentions: Human recombinant COX-2 was overexpressed in Sf9 insect cells (American Type Culture Collection). The microsomal fractions (∼8 μl) were suspended in Tris (100 mM, pH 8.0) as in reference 35. ASA was incubated (∼2 mM, 37°C, 30 min) with COX-2 before addition of DHA (10 μM), or in some experiments [1-14C]-labeled DHA (American Radiolabeled Chemicals, Inc.), and conversions were monitored in parallel using [1-14C]-labeled C20:4 (with ASA) (as in Fig. 2 ; also see text).

Bottom Line: Human neutrophils transformed COX-2-ASA-derived 17R-hydroxy-DHA into two sets of novel di- and trihydroxy products; one initiated via oxygenation at carbon 7 and the other at carbon 4.These compounds inhibited (IC(50) approximately 50 pM) microglial cell cytokine expression and in vivo dermal inflammation and peritonitis at ng doses, reducing 40-80% leukocytic exudates.These results indicate that exudates, vascular, leukocytes and neural cells treated with aspirin convert DHA to novel 17R-hydroxy series of docosanoids that are potent regulators.

View Article: PubMed Central - PubMed

Affiliation: Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesiology, Perioperative, and Pain Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA. cnserhan@zeus.bwh.harvard.edu

ABSTRACT
Aspirin (ASA) is unique among current therapies because it acetylates cyclooxygenase (COX)-2 enabling the biosynthesis of R-containing precursors of endogenous antiinflammatory mediators. Here, we report that lipidomic analysis of exudates obtained in the resolution phase from mice treated with ASA and docosahexaenoic acid (DHA) (C22:6) produce a novel family of bioactive 17R-hydroxy-containing di- and tri-hydroxy-docosanoids termed resolvins. Murine brain treated with aspirin produced endogenous 17R-hydroxydocosahexaenoic acid as did human microglial cells. Human COX-2 converted DHA to 13-hydroxy-DHA that switched with ASA to 17R-HDHA that also proved a major route in hypoxic endothelial cells. Human neutrophils transformed COX-2-ASA-derived 17R-hydroxy-DHA into two sets of novel di- and trihydroxy products; one initiated via oxygenation at carbon 7 and the other at carbon 4. These compounds inhibited (IC(50) approximately 50 pM) microglial cell cytokine expression and in vivo dermal inflammation and peritonitis at ng doses, reducing 40-80% leukocytic exudates. These results indicate that exudates, vascular, leukocytes and neural cells treated with aspirin convert DHA to novel 17R-hydroxy series of docosanoids that are potent regulators. These biosynthetic pathways utilize omega-3 DHA and EPA during multicellular events in resolution to produce a family of protective compounds, i.e., resolvins, that enhance proresolution status.

Show MeSH
Related in: MedlinePlus