Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages
Red clover (<i>Trifolium pratense</i>) possesses various dietary compounds that improve human health. However, the functions of anthocyanins in red clover remain unclear. Here we examined anti-inflammatory and antioxidant effects of red clover extract (RC) and red clover anthocyanins fra...
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doaj-5c2e80d196ae4d7fad728bf0f488cff72020-11-25T01:45:56ZengMDPI AGNutrients2072-66432020-04-01121089108910.3390/nu12041089Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 MacrophagesSang Gil Lee0Cindi R. Brownmiller1Sun-Ok Lee2Hye Won Kang3Department of Food Science and Nutrition, Pukyong National University, Busan 48513, KoreaDepartment of Food Science, University of Arkansas, Fayetteville, AR 72704, USADepartment of Food Science, University of Arkansas, Fayetteville, AR 72704, USAFood and Nutritional Sciences, Department of Family and Consumer Sciences, North Carolina Agricultural and Technical State University, Greensboro, NC 27411, USARed clover (<i>Trifolium pratense</i>) possesses various dietary compounds that improve human health. However, the functions of anthocyanins in red clover remain unclear. Here we examined anti-inflammatory and antioxidant effects of red clover extract (RC) and red clover anthocyanins fraction (RCA) using lipopolysaccharide (LPS)-treated RAW 264.7 macrophages and identified dietary compounds. RC and RCA suppressed LPS-induced expression of genes such as tumor necrosis factor (<i>TNF</i>)<i>α</i>, interleukin (<i>IL</i>)<i>1β</i>, inducible nitric oxide synthase (<i>iNOS</i>), monocyte chemoattractant protein (<i>MCP</i>)1, and cyclooxygenase (<i>COX</i>)<i>2</i>. LPS-stimulated intracellular reactive oxygen species (ROS) production also was prevented by both RC and RCA. NADPH oxidase 1 (NOX1) gene and phosphorylation of p47<sup>phox</sup> of NOX1 that were increased by LPS were inhibited in the cells treated with RCA. LPS-stimulated nuclear factor erythroid 2-related factor 2 (<i>NRF2</i>) gene expression and nuclear translocation of nuclear factor kappa B (NF-kB) subunit p65 were suppressed together with reduced iNOS and COX2 proteins by RCA. Additionally, 27 polyphenols and 7 anthocyanins from RC were identified and quantified. In conclusion, RC, especially RCA, exerted anti-inflammatory and anti-oxidative activities in vitro by regulating NF-κB and NRF2 signaling pathways, suggesting that anthocyanins in red clover are the potential candidates to reduce inflammation and oxidative stress.https://www.mdpi.com/2072-6643/12/4/1089red cloveranthocyaninsanti-inflammationantioxidation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sang Gil Lee Cindi R. Brownmiller Sun-Ok Lee Hye Won Kang |
spellingShingle |
Sang Gil Lee Cindi R. Brownmiller Sun-Ok Lee Hye Won Kang Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages Nutrients red clover anthocyanins anti-inflammation antioxidation |
author_facet |
Sang Gil Lee Cindi R. Brownmiller Sun-Ok Lee Hye Won Kang |
author_sort |
Sang Gil Lee |
title |
Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages |
title_short |
Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages |
title_full |
Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages |
title_fullStr |
Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages |
title_full_unstemmed |
Anti-Inflammatory and Antioxidant Effects of Anthocyanins of <i>Trifolium pratense</i> (Red Clover) in Lipopolysaccharide-Stimulated RAW-267.4 Macrophages |
title_sort |
anti-inflammatory and antioxidant effects of anthocyanins of <i>trifolium pratense</i> (red clover) in lipopolysaccharide-stimulated raw-267.4 macrophages |
publisher |
MDPI AG |
series |
Nutrients |
issn |
2072-6643 |
publishDate |
2020-04-01 |
description |
Red clover (<i>Trifolium pratense</i>) possesses various dietary compounds that improve human health. However, the functions of anthocyanins in red clover remain unclear. Here we examined anti-inflammatory and antioxidant effects of red clover extract (RC) and red clover anthocyanins fraction (RCA) using lipopolysaccharide (LPS)-treated RAW 264.7 macrophages and identified dietary compounds. RC and RCA suppressed LPS-induced expression of genes such as tumor necrosis factor (<i>TNF</i>)<i>α</i>, interleukin (<i>IL</i>)<i>1β</i>, inducible nitric oxide synthase (<i>iNOS</i>), monocyte chemoattractant protein (<i>MCP</i>)1, and cyclooxygenase (<i>COX</i>)<i>2</i>. LPS-stimulated intracellular reactive oxygen species (ROS) production also was prevented by both RC and RCA. NADPH oxidase 1 (NOX1) gene and phosphorylation of p47<sup>phox</sup> of NOX1 that were increased by LPS were inhibited in the cells treated with RCA. LPS-stimulated nuclear factor erythroid 2-related factor 2 (<i>NRF2</i>) gene expression and nuclear translocation of nuclear factor kappa B (NF-kB) subunit p65 were suppressed together with reduced iNOS and COX2 proteins by RCA. Additionally, 27 polyphenols and 7 anthocyanins from RC were identified and quantified. In conclusion, RC, especially RCA, exerted anti-inflammatory and anti-oxidative activities in vitro by regulating NF-κB and NRF2 signaling pathways, suggesting that anthocyanins in red clover are the potential candidates to reduce inflammation and oxidative stress. |
topic |
red clover anthocyanins anti-inflammation antioxidation |
url |
https://www.mdpi.com/2072-6643/12/4/1089 |
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