• Journal of Life Science
• /
• v.24 no.9
• /
• pp.973-980
• /
• 2014

This study was conducted to explore new nutraceutical resources from the plant kingdom possessing biological activities. To fulfill this purpose, the anti-oxidative and anti-inflammatory activities of Decaisnea insignis ethanol extract (DIEE) were evaluated. First, DIEE possessed potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar to ascorbic acid used as a positive control. Moreover, DIEE inhibited lipopolysaccharide (LPS)- and hydrogen peroxide ($H_2O_2$)-induced reactive oxygen species (ROS) in RAW 264.7 cells. Furthermore, DIEE induced the expression of an anti-oxidative enzyme, heme oxygenase 1 (HO-1), and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), in a dose-dependent manner. The modulation of the HO-1 and Nrf2 expressions might be regulated by mitogen-activated protein kinases (MAPKs) and their upstream signaling pathways. On the other hand, DIEE suppressed LPS-induced nitric oxide (NO) formation without cytotoxicity. The inhibition of the NO formation was the result of the downregulation of inducible NO synthase (iNOS) by DIEE. The suppression of NO and iNOS by DIEE might be modulated by their upstream transcription factors, nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), and activator protein 1 (AP-1) pathways. Taken together, these results provide important new insights that D. insignis possesses anti-oxidative and anti-inflammatory activities. Therefore, it might be utilized as a promising material in the field of nutraceuticals.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2019.10a
• /
• pp.92-92
• /
• 2019

Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.

• Journal of Applied Biological Chemistry
• /
• v.62 no.1
• /
• pp.39-50
• /
• 2019

Mitogen-activated protein (MAP) kinases play an important role in cell growth and differentiation, as well as the modulation of proinflammatory cytokines. The objective of this study was to examine the increase in the anti-inflammatory effect of Gentiana scabra Bunge (GSB), due to bioconversion with the Aspergillus kawachii crude enzyme, via inhibition of the $NF-{\kappa}B$ signaling and MAP kinase pathways in RAW 264.7 cells. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 in RAW 264.7 cells treated with the GSB ethyl acetate fraction bioconverted with A. kawachii crude enzyme (GE-BA), was dramatically suppressed as compared to GSB ethyl acetate fraction non-bioconverted with the A. kawachii crude enzyme (GE-UA). The phosphorylation of p38, extracellular signal-regulated kinases, and inhibitory ${\kappa}B$ in RAW 264.7 cells treated with GE-BA was further suppressed, as compared to exposure to GE-UA. Moreover, the mRNA expression of interleukin 6, interleukin 1-beta, and tumor necrosis $factor-{\alpha}$ was further suppressed by GE-BA, compared to GE-UA. Similarly, anti-oxidant activities, such as 2,2-diphenyl-1-picrylhydrazyl hydrate and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity, of GE-BA were further increased compared to GE-UA. These observations demonstrate that the anti-oxidant and anti-inflammatory activities of GSB ethyl acetate fraction increases as a result from bioconversion with the A. kawachii crude enzyme.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.54 no.2
• /
• pp.152-161
• /
• 2021

Eisenia bicyclis is known to have secondary metabolites exhibiting various biological activities. In a preliminary study, the n-hexane fraction obtained from the ethanolic extract of E. bicyclis showed higher anti-inflammatory activity than the ethyl acetate and butyl alcohol fractions based on the inhibition of lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in RAW 264.7 cells. Using this fraction (E. bicyclis hexane fraction, EHF), we investigated the molecular mechanisms underlying its anti-inflammatory effect in LPS-stimulated RAW 264.7 cells. Pretreatment of the cells with up to 50 ㎍/mL EHF significantly inhibited NO and prostaglandin E₂ production as well as their responsible enzyme proteins and mRNAs, in a dose-dependent manner (P<0.05). Similarly, EHF markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α as well as their mRNA levels. Nuclear translocation of nuclear factor-kappa B (NF-κB) was strongly suppressed by EHF treatment. EHF significantly reduced the phosphorylation of mitogen-activated protein kinases and phosphatidylinositol 3-kinase/Akt in LPS-stimulated cells. Moreover, EHF reduced ear edema in phorbol myristate acetate (PMA)-induced mice. These results indicate that EHF contains potent anti-inflammatory compounds, which may be used as a dietary supplement for the prevention of inflammatory diseases.

• Natural Product Sciences
• /
• v.23 no.1
• /
• pp.16-20
• /
• 2017

In a continuing study of the fermented noni (Morinda citrifolia) juice exudates, five compounds, heptanyl $2-O-{\beta}-{\small{D}}-xylofuranosyl-(1{\rightarrow}6)-{\beta}-{\small{D}}-glucopyranoside$ (1), n-butyl ${\beta}-{\small{D}}-glucopyranoside$ (2), (1S)-(3-ethenyl-phenyl)-1,2-ethanediol (3), (2S)-2-hydroxybutanedioic acid (4), and daucosterol (5) were isolated from the buthanol partition of the extract. The structures of the isolates were identified by 1D and 2D NMR, and MS experiments, as well as by comparison of their data with the published values. Among the isolates, compounds 1 - 3 were isolated for the first time from the plant species. The isolated compounds were evaluated for their cancer chemopreventive potential based on their ability to inhibit nitric oxide (NO) production and tumor necrosis factor alpha ($TNF-{\alpha}$)-induced $NF-{\kappa}B$ activity, and quinonone reductase-1 (QR1)-inducing effect.

• The Journal of Korean Medicine
• /
• v.27 no.1 s.65
• /
• pp.184-195
• /
• 2006

Objectives : Betula Platyphylla(BP) is a traditional analgesic, anti-fungal, anti-inflammatory herb used in Chinese 1medicine. However, no information is available to explain its action. In this study. we investigated the anti-inflammatory 1effects of BP to elutidate the molecular pharmacological activity in the ethanol extract of BP(BPE). Methods : We performed WTS assay in lipopolysaccharide (LPS) stimulated RAW264.7 macrophages with BPE. Nitrite was measured by Griess assay, prostaglandin E2 (PGE2) by enzyme-linked immunosorbent assay (ELISA) in LPS induced RAW264.7 macrophages with BPE. Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) were determined by Western blot. Activation of nuclear factor-kappaB (NF-kB) was measured by electrophoretic mobility shift assay (EMSA). Results : BPE significantly suppressed production of nitric oxide (NO) and PGE2 in LPS-induced RAW264.7 macrophages in a dose-dependent manner. The maximal inhibition rate of NO and PGE2 production by BPE was ca. 88.8% and 93% at the concentration of $100{\mu}g/ml$ (non-cytotoxic concentration), respectively. BPE also decreased iNOS protein and COX-2 protein in LPS-induced RAW264.7 macrophages. EMSA demonstrated that BPE inhibited the DNA binding activity of the NF-kB. Conclusions : These results suggest that BPE inhibits NF-${\kappa}B$-mediated gene expression and downregulates inflammatory mediator production in RAW264.7 macrophages.

• Korean Journal of Organic Agriculture
• /
• v.32 no.1
• /
• pp.91-105
• /
• 2024

Plantago asiatica L. (P. asiatica) is a perennial plant belonging to the plantaginaceae and is useful in treating a various diseases such as wounds, bronchitis, and chronic constipation. The bioactive effects of P. asiatica extract was evaluated to determine its potential for use as a variety materials in the food, pharmaceutical, and agricultural industries. Polyphenol and flavonoid contents, free radical scavenging, reducing power activity, and reactive oxygen species (ROS) expression were measured to identify the antioxidative activity. Anti-inflammatory effects were evaluated via analysis of nitric oxide (NO) and pro-inflammatory protein expression in LPS-induced RAW 264.7 cell. As a result of measuring the antioxidant activities of the P. asiatica extract, the total polyphenol content was 50.91±0.78 mg gallic acid equivalents/g and the flavonoid content was 100.99±0.44 mg rutin equivalents/g, and both DPPH and ABTS radical scavenging activities and reducing power increased depending on the concentration. Also, intracellular ROS production was inhibited by the P. asiatica extract. No cytotoxicity was observed when P. asiatica extract was treated, and NO and inflammatory protein expression were inhibited, and nuclear factor kappa B (NF-κB) phosphorylation was also inhibited in a concentration-dependent manner. In conclusion, P. asiatica is a functional natural resources of antioxidant and anti-inflammatory agents that can be used in various industries, including food and agriculture.

• Journal of Life Science
• /
• v.28 no.11
• /
• pp.1361-1368
• /
• 2018

Inflammation is the most common condition in the human body. Tissue damage triggers inflammation, together with vasodilation and increased blood flow at the inflamed site, resulting in edema. Inflammatory responses are also triggered by lipopolysaccharide (LPS), a Toll-like receptor Enterococcus faecalis, a gram-positive organism, has been reported to possess immunomodulatory and preventive activities; however, its use may present risks of sepsis and other systemic infections. Heat-killed Enterococcus faecalis (EF-2001) has been reported to induce antitumor activity, but its effects on inflammation are not known. In the present study, we investigated the effect of EF-2001 on LPS-induced macrophage inflammatory responses. EF-2001 treatment reduced nitric oxide (NO) production, indicating suppression of inflammatory reactions. EF-2001 showed no cytotoxicity in macrophages. Further investigation of the anti-inflammatory mechanism of EF-2001 indicated that EF-2001 reduced the LPS-induced expression of inducible nitric oxide synthase and cyclooxygenase-2. EF-2001 also reduced f the LPS induction of several inflammatory molecules involved in the nuclear factor-${\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinase pathways, including ERK, JNK, and p38 phosphorylation, in a concentration-dependent manner. Additionally, EF-2001 inhibited Akt phosphorylation and increased the expression of the inhibitory ${\kappa}B$ ($I{\kappa}B$) protein, an inhibitor of $NF-{\kappa}B$. EF-2001 also inhibited the nuclear translocation of p65. These results suggest that EF-2001 has anti-inflammatory properties and may be useful for treating inflammatory diseases.

• Natural Product Sciences
• /
• v.17 no.2
• /
• pp.135-141
• /
• 2011

Celastrus orbiculatus has been widely used as a traditional medicine for the treatment of many diseases including rheumatoid arthritis and odontalgia. In the present study, anti-metastatic activity of a methanolic extract from C. orbiculatus (MCO) was studied. A gelatin zymographic assay revealed that MCO has potent inhibitory effects on MMP-2 and MMP-9 activities in B16F10 melanoma cells. Moreover, MCO attenuated MMP expression via down-regulation of NF-${\kappa}$B translocation to the nucleus. Melanoma cell migration and invasion were also down-regulated by MCO. In addition, MCO significantly suppressed lung metastasis in an in vivo model. These results strongly suggest that MCO may possibly be used as a valuable anti-metastatic agent for cancer treatment.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.44 no.4
• /
• pp.532-539
• /
• 2015

Inhibition of osteoclast differentiation is the most important target for prevention of inflammatory bone resorption and bone diseases. Here, we investigated the effect of spinach ethanol extract on osteoclast differentiation in RAW264.7 cells. Spinach was extracted with ethanol at a concentration ranging from 0 to 100% (0, 25, 50, 75, and 100% ethanol). Inhibitory effects of receptor activator of NF-${\kappa}B$ ligan (RANKL)-induced osteoclast differentiation were evaluated using tartrate-resistant acid phosphatase (TRAP) stain assay. The most effective eanol concentration for osteoclast differentiation was 100%. Spinach extract (100% ethanol) suppressed RANKL-induced osteoclast differentiation and TRAP activity. Spinach extract (100% ethanol) also suppressed expression of osteoclast differentiation-related marker genes (NFATc1, c-FOS, cathepsin K, and TRAP) and down-regulated RANKL-induced NF-${\kappa}B$ and ERK phosphorylation during osteoclast differentiation. Taken together, our results suggest that spinach extract is effective against reducing osteoclast differentiation through the NF-${\kappa}B$-mediated pathway.