Naringenin suppresses Toll‐like receptor 2‐mediated inflammatory responses through inhibition of receptor clustering on lipid rafts

Kataoka, Hideo; Saeki, Ayumi; Hasebe, Akira; Shibata, Kenichiro; Into, Takeshi

doi:10.1002/fsn3.2063

Cited by 8 publications

(6 citation statements)

References 55 publications

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“…The human monocytic cell line THP-1, obtained from RIKEN BioResource Research Center, was cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, penicillin G (100 units/ml) and streptomycin (100 μg/ml) in a humidified atmosphere containing 5% CO 2 at 37°C, as described previously [ 11 ]. The cells were differentiated into macrophage-like attached cells by incubation with 20 nM phorbol 12-myristate 13-acetate (Sigma) for 24 h. After a medium change, the cells were further incubated for 24 h and used for treatment with the peptide solution in a serum-free medium.…”

Section: Methodsmentioning

confidence: 99%

Solubility affects IL-1β-producing activity of the synthetic candidalysin peptide

et al. 2022

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Candidalysin, a peptide toxin produced specifically from hyphae of Candida albicans, plays a crucial role in C. albicans pathogenesis in the oral cavity and vagina. Synthetic peptides have been widely used in previous studies to investigate the bioactivity of candidalysin. Although the solubility of the peptide, which is expected to have a hydrophobic property, has not been well characterized, candidalysin solutions are usually prepared in water. In this study, we prepared the synthetic peptide candidalysin in water (CLw) or in dimethyl sulfoxide (CLd) and compared their cytotoxicity and interleukin (IL)-1β-producing activity to determine whether the activity of the peptide would be affected. In addition, we evaluated whether the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome pathway or other pathways were involved in their activities. Unexpectedly, we found that CLw was not completely solubilized and contained abundant insoluble microparticles. CLw was active at comparably high concentrations (≥ 10 μM). In contrast, CLd is completely solubilized and sufficiently active at low concentrations, that is, 1 μM or less. CLw showed weak cytotoxicity and NLRP3-dependent and cathepsin B-dependent IL-1β-producing activity, whereas CLd showed strong cytotoxicity and cathepsin B-dependent IL-1β-producing activity. Fractionation of CLw revealed that NLRP3-dependent activity was caused by insoluble microparticles. Furthermore, nanoparticle tracking of CLd revealed that the peptide was present as nanoparticles with a size of 96 nm. CLw contained a small amount of such nanoparticles. Thus, the bioactivities of the synthetic peptide candidalysin, especially the IL-1β-producing activity, are affected by the solubility of the peptide depending on the solvent employed. The NLRP3-dependent activity of the synthetic peptide is caused by insoluble microparticles and may not be the intrinsic activity of candidalysin.

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Section: Methodsmentioning

confidence: 99%

Solubility affects IL-1β-producing activity of the synthetic candidalysin peptide

et al. 2022

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“…In addition, plant-and seaweedderived polysaccharides activate signal transduction and cytokine production downstream of TLR2 (26,28). The citrus flavonoid, naringenin, has been reported to suppress TLR2 signaling (37,38). Based on these studies, low molecular-weight polyphenols, such as plant-derived flavonoids, may act by suppressing TLR2 and TLR4, whereas plant-derived polysaccharides activate TLR4.…”

Section: Effect Of Kumaizasa Leaf Extract (Kle) On Lipopolysaccharide...mentioning

confidence: 99%

“…TLR-mediated immunomodulation is very important in cancer and inflammatory disease therapies. Notably, chronic inflammatory conditions induce carcinogenesis and cancer cachexia, and modulating immunity through naturally-occurring compounds, such as KLE, is desirable (38)(39)(40). However, regulatory effects of KLE on TLR2 and TLR4 and specific components involved remain unclear.…”

Section: Effect Of Kumaizasa Leaf Extract (Kle) On Lipopolysaccharide...mentioning

confidence: 99%

Effects of Kumaizasa (Sasa senanensis) Leaf Extract on Innate Immune Regulation in HEK293 Cells and Macrophages

et al. 2021

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Background/Aim: We investigated the effect of Kumaizasa leaf extract (KLE) on innate immunity using the HEK293 and RAW 264.7 cell lines. Materials and Methods: KLE, lipopolysaccharides (LPS), or KLE with LPS were added to RAW 264.7 cells. The TNF-α and IL-1β mRNA expression was then quantified. The expression of MAPKs, NFĸB, TNF-α and IL-1β proteins was also quantified. In addition, KLE was added to HEK293 cells and the IL-8 concentration was measured. Results: In RAW 264.7 cells, KLE increased the levels of TNF-α and IL-1β mRNA. By contrast, when KLE and LPS were added to RAW 264.7 cells, the increase in TNF-α and IL-1β mRNA was ameliorated. Similarly, the expression of JNK and ERK proteins was reduced. The addition of KLE to HEK293 cells induced IL-8 production. Conclusion: Based on these results, a KLE-mediated mechanism may regulate immunity by suppressing the expression of JNK and ERK, which are involved in inflammatory signal transduction.The Kumaizasa (Sasa senanensis) plant belongs to the Gramineae family and is widely found in northern Japan. Kumaizasa consists primarily of polysaccharides; however, low molecular weight compounds have also been identified, including p-coumaric acid, ferulic acid, vanillin, apigenin, luteolin, tricin, syringaresinol, phenylpropanoid, phydroxybenzoic acid, p-hydroxybenzaldehyde, and 3,4dihydroxybenzaldehyde (1, 2).Sasa, including Kumaizasa, has been reported to act as a UV protectant (3), antiviral (4), antioxidant (in a rat small intestinal ischemia-reperfusion model) (5), as a suppressor of amyloid-β-mediated neuronal damage (6, 7), and as a suppressor of macrophage-derived nitric oxide (NO) (8) and prostaglandin E 2 (PGE 2 ) production (9). Other effects include antitumor activity in mouse K562 (human chronic myeloid leukemia) and YAC1 (murine lymphoma) cells, inhibition of histamine release in rat intraperitoneal cells, and improvement of the oral and intestinal environment (10).Our understanding of the innate immune system, inflammation and biological defense systems has progressed significantly, especially with respect to the discovery of tolllike receptors (TLRs). TLRs are expressed in many types of cells and play important roles in macrophages and dendritic cells of the innate immune system. TLRs recognize foreign substances and stimulate the production of immunomodulatory substances, such as cytokines, through intracellular signaling. These cytokines have direct antibacterial, antiviral and antitumor effects and play various roles, including the regulation of innate and adaptive immune systems (11)(12)(13).The TLR family comprises 11 members. β-Glucan, which is derived from the cell wall of bacteria, is a ligand for TLR2, whereas lipopolysaccharide (LPS), derived from gram-negative bacteria, is a ligand for TLR4. Inflammatory cytokines, including TNF-α, IL-8, IL-1β, are produced following the stimulation of TLR2 and TLR4. TLR signaling activates mitogen-activated protein kinase (MAPK), c-Jun Nterminal kinase (c-JNK), extracellular signal-regulated kinase (ERK), p3...

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“…20,21 METAFECTENE® PRO exhibits high transfection e ciency and low toxicity in multiple cell lines and/or primary cells including, HEK 293T cells, THP-1 (RCB1189), BV2 cells, primary T-cells and Jurkat; HEK293 cells. 20,[22][23][24][25][26] Lipofectamine™ LTX with PLUS™ reagent is an origin-free liposomal transfection reagent. Lipofectamine is effective, easy to use and relatively less expensive compared to the other methods and generally achieve relatively high transfection e ciency.…”

Section: Introductionmentioning

confidence: 99%

Comparison and optimization of transfection methods for human lymphoblast TK6 cell line

Ewunkem

Agee

2022

Preprint

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Transfection has recently gained attention in the field of biomedical research due to its ability to manipulate gene expression. Every mammalian cell type has a characteristic set of requirements for optimal transfection. Some cells can be difficult to transfect and requires optimization for successful transfection. Human lymphoblast TK6 cell line, an important cell line for genotoxic studies, are known to be extremely hard to transfect. Transfection methods for human lymphoblast TK6 is increasingly important. To accomplish this, TK6 human lymphoblasts were transfected with plasmid constructs that expressed Green fluorescent Protein (GFP) and NanoLuc® activity. We compared the transfection efficiencies of three commercially available transfection reagents including Amaxa 96-well Nucleofection procedure using various solutions (SF, SE and SG), Lipofectamine LTX, and Metafectene Pro®. The transfection efficiencies, and toxicity of various reagents were tested by fluorescence microscopy, luciferase activity, and cell viability assays. Amaxa 96-well Nucleofection Solution SF was identified as the best transfection reagent due to its relatively high luciferase activity, acceptable cell viability (80%), and GFP transfection efficiency (80%). Efficient conditions for transfection utilized with this reagent included 0.4µg of plasmid DNA, 1.8 x 105 cells, and the DS 137 nucleofector program.

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Naringenin suppresses Toll‐like receptor 2‐mediated inflammatory responses through inhibition of receptor clustering on lipid rafts

Cited by 8 publications

References 55 publications

Solubility affects IL-1β-producing activity of the synthetic candidalysin peptide

Solubility affects IL-1β-producing activity of the synthetic candidalysin peptide

Effects of Kumaizasa (Sasa senanensis) Leaf Extract on Innate Immune Regulation in HEK293 Cells and Macrophages

Comparison and optimization of transfection methods for human lymphoblast TK6 cell line

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