An Optimized Ex Vivo n–3 PUFA Supplementation Strategy for Primary Human Macrophages Shows That DHA Suppresses Prostaglandin E2 Formation
ABSTRACT Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n–3 PUFA supplementation strategy. M2‐like macrophages were supple...
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| Published in | Molecular nutrition & food research Vol. 69; no. 1; pp. e202400716 - n/a |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Germany
Wiley Subscription Services, Inc
01.01.2025
John Wiley and Sons Inc |
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| Online Access | Get full text |
| ISSN | 1613-4125 1613-4133 1613-4133 |
| DOI | 10.1002/mnfr.202400716 |
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| Abstract | ABSTRACT
Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n–3 PUFA supplementation strategy. M2‐like macrophages were supplemented for 2–3 days with 20–40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA‐preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n–3 PUFA <0.25 mM for the selection of donors of plasma as well as %n–6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n–3 and lower n–6 PUFAs. This was accompanied by a decrease of arachidonic acid‐derived oxylipins in a dose‐ and time‐dependent manner in favor of n–3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro‐inflammatory prostaglandins in the DHA‐supplemented cells, but no changes in cytokines. In vitro supplementation studies with n–3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n–3 PUFAs in primary human immune cells, offering an alternative for intervention studies.
This presented ex vivo n–3 polyunsaturated fatty acid (PUFA) supplementation strategy is an important tool for the mechanistic investigation of n–3 PUFA mediated effects in human immune cells. Fatty acid (FA) pattern from monocytes derived from subjects with a low n–3 PUFA status was shifted toward one reflecting the pattern of subjects with a high n–3 PUFA status, accompanied by remarkable changes in the oxylipin profile. Lower levels of pro‐inflammatory prostaglandins were found in the supplemented macrophages, which may be one of the major mechanisms of how the anti‐inflammatory effect of n–3 PUFA is mediated. |
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| AbstractList | Evidence suggests beneficial effects of long-chain n-3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n-3 PUFA supplementation strategy. M2-like macrophages were supplemented for 2-3 days with 20-40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA-preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n-3 PUFA <0.25 mM for the selection of donors of plasma as well as %n-6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n-3 and lower n-6 PUFAs. This was accompanied by a decrease of arachidonic acid-derived oxylipins in a dose- and time-dependent manner in favor of n-3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro-inflammatory prostaglandins in the DHA-supplemented cells, but no changes in cytokines. In vitro supplementation studies with n-3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n-3 PUFAs in primary human immune cells, offering an alternative for intervention studies.Evidence suggests beneficial effects of long-chain n-3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n-3 PUFA supplementation strategy. M2-like macrophages were supplemented for 2-3 days with 20-40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA-preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n-3 PUFA <0.25 mM for the selection of donors of plasma as well as %n-6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n-3 and lower n-6 PUFAs. This was accompanied by a decrease of arachidonic acid-derived oxylipins in a dose- and time-dependent manner in favor of n-3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro-inflammatory prostaglandins in the DHA-supplemented cells, but no changes in cytokines. In vitro supplementation studies with n-3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n-3 PUFAs in primary human immune cells, offering an alternative for intervention studies. Evidence suggests beneficial effects of long‐chain n –3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n –3 PUFA supplementation strategy. M2‐like macrophages were supplemented for 2–3 days with 20–40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA‐preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n –3 PUFA <0.25 mM for the selection of donors of plasma as well as % n –6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n –3 and lower n –6 PUFAs. This was accompanied by a decrease of arachidonic acid‐derived oxylipins in a dose‐ and time‐dependent manner in favor of n –3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro‐inflammatory prostaglandins in the DHA‐supplemented cells, but no changes in cytokines. In vitro supplementation studies with n –3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n –3 PUFAs in primary human immune cells, offering an alternative for intervention studies. Evidence suggests beneficial effects of long-chain n-3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n-3 PUFA supplementation strategy. M2-like macrophages were supplemented for 2-3 days with 20-40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA-preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n-3 PUFA <0.25 mM for the selection of donors of plasma as well as %n-6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n-3 and lower n-6 PUFAs. This was accompanied by a decrease of arachidonic acid-derived oxylipins in a dose- and time-dependent manner in favor of n-3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro-inflammatory prostaglandins in the DHA-supplemented cells, but no changes in cytokines. In vitro supplementation studies with n-3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n-3 PUFAs in primary human immune cells, offering an alternative for intervention studies. ABSTRACT Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n–3 PUFA supplementation strategy. M2‐like macrophages were supplemented for 2–3 days with 20–40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA‐preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n–3 PUFA <0.25 mM for the selection of donors of plasma as well as %n–6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n–3 and lower n–6 PUFAs. This was accompanied by a decrease of arachidonic acid‐derived oxylipins in a dose‐ and time‐dependent manner in favor of n–3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro‐inflammatory prostaglandins in the DHA‐supplemented cells, but no changes in cytokines. In vitro supplementation studies with n–3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n–3 PUFAs in primary human immune cells, offering an alternative for intervention studies. This presented ex vivo n–3 polyunsaturated fatty acid (PUFA) supplementation strategy is an important tool for the mechanistic investigation of n–3 PUFA mediated effects in human immune cells. Fatty acid (FA) pattern from monocytes derived from subjects with a low n–3 PUFA status was shifted toward one reflecting the pattern of subjects with a high n–3 PUFA status, accompanied by remarkable changes in the oxylipin profile. Lower levels of pro‐inflammatory prostaglandins were found in the supplemented macrophages, which may be one of the major mechanisms of how the anti‐inflammatory effect of n–3 PUFA is mediated. Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n–3 PUFA supplementation strategy. M2‐like macrophages were supplemented for 2–3 days with 20–40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA‐preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n–3 PUFA <0.25 mM for the selection of donors of plasma as well as %n–6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n–3 and lower n–6 PUFAs. This was accompanied by a decrease of arachidonic acid‐derived oxylipins in a dose‐ and time‐dependent manner in favor of n–3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro‐inflammatory prostaglandins in the DHA‐supplemented cells, but no changes in cytokines. In vitro supplementation studies with n–3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n–3 PUFAs in primary human immune cells, offering an alternative for intervention studies. Evidence suggests beneficial effects of long‐chain n –3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n –3 PUFA supplementation strategy. M2‐like macrophages were supplemented for 2–3 days with 20–40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA‐preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n –3 PUFA <0.25 mM for the selection of donors of plasma as well as % n –6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n –3 and lower n –6 PUFAs. This was accompanied by a decrease of arachidonic acid‐derived oxylipins in a dose‐ and time‐dependent manner in favor of n –3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro‐inflammatory prostaglandins in the DHA‐supplemented cells, but no changes in cytokines. In vitro supplementation studies with n –3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n –3 PUFAs in primary human immune cells, offering an alternative for intervention studies. This presented ex vivo n –3 polyunsaturated fatty acid (PUFA) supplementation strategy is an important tool for the mechanistic investigation of n –3 PUFA mediated effects in human immune cells. Fatty acid (FA) pattern from monocytes derived from subjects with a low n –3 PUFA status was shifted toward one reflecting the pattern of subjects with a high n –3 PUFA status, accompanied by remarkable changes in the oxylipin profile. Lower levels of pro‐inflammatory prostaglandins were found in the supplemented macrophages, which may be one of the major mechanisms of how the anti‐inflammatory effect of n –3 PUFA is mediated. |
| Author | Kampschulte, Nadja Rothweiler, Carina Kirchhoff, Rebecca Schebb, Nils Helge Rohwer, Nadine Weylandt, Karsten‐Henrich |
| AuthorAffiliation | 3 Faculty of Health Sciences, Joint Faculty of the Brandenburg University of Technology Brandenburg Medical School and University of Potsdam Potsdam Germany 2 Division of Medicine, Department of Gastroenterology, Metabolism and Oncology University Hospital Ruppin‐Brandenburg, Brandenburg Medical School Neuruppin Germany 1 Chair of Food Chemistry, School of Mathematics and Natural Sciences University of Wuppertal Wuppertal Germany 4 Department of Molecular Toxicology German Institute of Human Nutrition Potsdam‐Rehbruecke, Nuthetal Germany |
| AuthorAffiliation_xml | – name: 3 Faculty of Health Sciences, Joint Faculty of the Brandenburg University of Technology Brandenburg Medical School and University of Potsdam Potsdam Germany – name: 4 Department of Molecular Toxicology German Institute of Human Nutrition Potsdam‐Rehbruecke, Nuthetal Germany – name: 1 Chair of Food Chemistry, School of Mathematics and Natural Sciences University of Wuppertal Wuppertal Germany – name: 2 Division of Medicine, Department of Gastroenterology, Metabolism and Oncology University Hospital Ruppin‐Brandenburg, Brandenburg Medical School Neuruppin Germany |
| Author_xml | – sequence: 1 givenname: Rebecca orcidid: 0009-0003-7196-9943 surname: Kirchhoff fullname: Kirchhoff, Rebecca organization: University of Wuppertal – sequence: 2 givenname: Nadja orcidid: 0000-0002-6253-9939 surname: Kampschulte fullname: Kampschulte, Nadja organization: University of Wuppertal – sequence: 3 givenname: Carina surname: Rothweiler fullname: Rothweiler, Carina organization: University of Wuppertal – sequence: 4 givenname: Nadine orcidid: 0000-0001-6170-1327 surname: Rohwer fullname: Rohwer, Nadine organization: German Institute of Human Nutrition – sequence: 5 givenname: Karsten‐Henrich orcidid: 0000-0002-5361-4327 surname: Weylandt fullname: Weylandt, Karsten‐Henrich organization: University Hospital Ruppin‐Brandenburg, Brandenburg Medical School – sequence: 6 givenname: Nils Helge orcidid: 0000-0003-1299-6629 surname: Schebb fullname: Schebb, Nils Helge email: nils@schebb-web.de organization: University of Wuppertal |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/39676434$$D View this record in MEDLINE/PubMed |
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| Keywords | arachidonic acid cascade lipid mediators oxylipins fatty acids inflammation |
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| Snippet | ABSTRACT
Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying... Evidence suggests beneficial effects of long‐chain n –3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are... Evidence suggests beneficial effects of long-chain n-3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are... Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are... |
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| StartPage | e202400716 |
| SubjectTerms | Adult Arachidonic acid arachidonic acid cascade blood Cells, Cultured cytokines Cytokines - metabolism Dietary Supplements Dinoprostone - metabolism Docosahexaenoic acid Docosahexaenoic Acids - pharmacology Fatty acids Fatty Acids, Omega-3 - pharmacology Female food research Humans Immune system inflammation Inflammatory diseases lipid mediators Lipopolysaccharides - pharmacology Macrophages Macrophages - drug effects Macrophages - metabolism Male nutrition omega-3 fatty acids omega-6 fatty acids oxylipins Oxylipins - pharmacology Polyunsaturated fatty acids Prostaglandin E2 Prostaglandins Supplements |
| Title | An Optimized Ex Vivo n–3 PUFA Supplementation Strategy for Primary Human Macrophages Shows That DHA Suppresses Prostaglandin E2 Formation |
| URI | https://onlinelibrary.wiley.com/doi/abs/10.1002%2Fmnfr.202400716 https://www.ncbi.nlm.nih.gov/pubmed/39676434 https://www.proquest.com/docview/3152012473 https://www.proquest.com/docview/3146851766 https://www.proquest.com/docview/3165873631 https://pubmed.ncbi.nlm.nih.gov/PMC11704825 |
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