Isotrifoliol inhibits pro-inflammatory mediators by suppression of TLR/NF-κB and TLR/MAPK signaling in LPS-induced RAW264.7 cells |
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| Institution: | 1. Industrial Bio-materials Research Center, KRIBB, Daejeon 34141, Republic of Korea;2. Department of Biomolecular Science, Korea University of Science and Technology, KRIBB, Daejeon 34141, Republic of Korea;3. College of Pharmacy, Chungnam National University, Daejeon 34134, Republic of Korea;4. Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Republic of Korea;5. Insect Biotech Co. Ltd., Daejeon 34054, Republic of Korea;1. Icahn School of Medicine at Mount Sinai, New York, NY 10029, United States;2. Hematology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, United States;3. Life Science and Technology, Changchun University of Science and Technology, Changchun 130022, China;1. Department of Life Science, Gachon University, 1342 Seongnamdaero, Seongnam 461-701, Republic of Korea;2. Jeju Biodiversity Research Institute, Jeju Technopark, Seogwipo 697-943, Republic of Korea;3. Department of Herbal Medicine Resources, Kangwon National University, Samcheok 245-710, Republic of Korea;4. School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea;5. Division of Information Analysis Research, Korea Institute of Science and Technology Information, KISTI, Seoul 130-741, Republic of Korea;6. Department of Food and Nutrition, Kangwon National University, Samcheok 245-710, Republic of Korea;7. Department of Physical Therapy, Kangwon National University, Samcheok 245-710, Republic of Korea;8. Department of Health Administration, College of Social Sciences, Joongbu University, Geumsan 312-702, Republic of Korea |
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| Abstract: | Soybeans, produced by Glycine max (L.) Merr., contain high levels of isoflavones, such as genistein and daidzein. However, soy leaves contain more diverse and abundant flavonol glycosides and coumestans, as compared to the soybean. This study investigated the anti-inflammatory effects of the major coumestans present in soy leaf (coumestrol, isotrifoliol, and phaseol) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Coumestans significantly reduced LPS-induced nitric oxide (NO), prostaglandin E2 (PGE2), and reactive oxygen species (ROS) production; isotrifoliol had the most potent anti-inflammatory activity. Isotrifoliol reduced LPS-mediated induction of mRNA expression of inducible nitric-oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin (IL)-1β, IL-6, tumor necrosis factor alpha (TNFα), and chemokines, such as chemokine (C-C motif) ligand (CCL) 2, CCL3, and CCL4. Isotrifoliol prevented NF-κB p65 subunit activation by reducing the phosphorylation and degradation of the inhibitor of NF-κB. And isotrifoliol significantly suppressed phosphorylation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (MAPK). Furthermore, isotrifoliol suppressed LPS-induced Toll-like Receptor (TLR) signaling pathway, including mRNA expression of TNF receptor associated factor 6, transforming growth factor beta-activated kinase 1 (TAK1), TAK1 binding protein 2 (TAB2), and TAB3. These results demonstrate that isotrifoliol exerts an anti-inflammatory effect by suppressing the expression of inflammatory mediators via inhibition of TLR/NF-κB and TLR/MAPK signaling in LPS-induced RAW264.7 macrophages. Therefore, isotrifoliol can be used as an anti-inflammatory agent, and coumestan-rich soy leaf extracts may provide a useful dietary supplement. |
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