• IMMUNE NETWORK
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• v.10 no.5
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• pp.145-152
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• 2010

Background: The flowers of Inula japonica (Inulae Flos) have long been used in traditional medicine for the treatment of inflammatory diseases. In the present study, we investigated the anti-inflammatory properties of Inulae Flos Extract (IFE). Methods: The anti-inflammatory effects of IFE against nitric oxide (NO), $PGE_2$, TNF-${\alpha}$, and IL-6 release, as well as NF-${\kappa}B$ and MAP kinase activation were evaluated in RAW 264.7 cells. Results: IFE inhibited the production of NO and the expression of inducible nitric oxide synthase (iNOS) in LPS-stimulated RAW264.7 cells. In addition, IFE reduced the release of pro-inflammatory cytokines, such as TNF-${\alpha}$ and IL-6. Furthermore, IFE inhibited the NF-${\kappa}B$ activation induced by LPS, which was associated with the abrogation of $I{\kappa}B-{\alpha}$ degradation and subsequent decreases in nuclear p65 and p50 levels. Moreover, the phosphorylation of ERK, JNK, and p38 MAP kinases in LPS-stimulated RAW 264.7 cells was suppressed by IFE in a dose-dependent manner. Conclusion: These results suggest that the anti-inflammation activities of IFE might be attributed to the inhibition of NO, iNOS and cytokine expression through the down-regulation of NF-${\kappa}B$ activation via suppression of $I{\kappa}B{\alpha}$ and MAP kinase phosphorylation in macrophages.

• Journal of Acupuncture Research
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• v.23 no.6
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• pp.103-115
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• 2006

Objectives : Rheumatoid arthritis(RA) is a systemic & a chronic inflammatory autoimmune disease . A chronic , locally destructive inflammmatory reaction in human is examplified by the synovitis present in some connective tissue disorder. The presence of a number of cytokines, $TNF-{\alpha}$, iNOS & expression of nitric oxide, NF-kB p65 activation implies an important role of cellular immune response in RA inflammatory reaction. This study was designed to evaluate on the effects of the Homnis Placenta herbal acupuncture on EX-LE201 & ST 35 reducing expression of LPS-induced arthritis model in mice. Materials and Methods : Homnis Placenta herbal acupuncture was inserted into 10 rats induced rheumatoid arthritis. The acupunctures were injected into the EX-LE201 and ST35 points. Such indexes were measured the inhibition of inducible nitric oxide synthase(iNOS) expression, nitric oxide(NO) production in vitro experiment and Tumor Necrosis $Factor-{\alpha}(TNF-{\alpha})$ & Nuclear Factor kappa $B(NF-{\kappa}B)$ p65 activation, synovial hyperplasia, angiogenesis and fibrosis in synovial membrane of knee joint of mice in vivo experiment. Results : 1.Homnis Placenta Herbal acupuncture inhibited iNOS mRNA and NO in RAW 264.7 cell of LPS-induced rheumatoid arthritis in a dose dependent manner. 2.Homnis Placenta Herbal acupuncture also showed significant inhibition of $TNF-{\alpha}$ & $NF-{\kappa}B$ p65, activation, synovial hyperplasia, angiogenesis and fibrosis in synovial membrane of knee joint of mice. Conclusion : These results suggest that Homnis Placenta Herbal acupuncture has an therapeutic effects on LPS induced-rheumatoid arthritis by inhibiting $TNF-{\alpha}$ activation.

• Molecular & Cellular Toxicology
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• v.5 no.4
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• pp.354-363
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• 2009

Cytotoxic Nitric oxide (NO) overproduced by inducible NO Synthase (iNOS or NOS2), which was induced in inflammatory reactions and immune responses directly or indirectly affects the functions as host defense and can cause normal tissue damage. Microarray analysis was performed to identify gene profiles of both NO-dependent and -independent transcripts in RAW 264.7 macrophages that use selective NOS2 inhibitors aminoguanidine ($100\;{\mu}M$) and L-canavanine (1 mM). A total of 3,297 genes were identified that were up- or down-regulated significantly over 2-fold in lipopolysaccharide (LPS)-treated macrophages. NO-dependency was determined in the expressed total gene profiles and also within inflammatory conditions-related functional categories. Out of all the gene profiles, 1711 genes affected NO-dependently and -independently in 567 genes. In the categories of inflammatory conditions, transcripts of 16 genes (Pomp, C8a, Ifih1, Irak1, Txnrd1, Ptafr, Scube1, Cd8a, Gpx4, Ltb, Fasl, Igk-V21-9, Vac14, Mbl1, C1r and Tlr6) and 29 geneas (IL-1beta, Mpa2l, IFN activated genes and Chemokine ligands) affected NO-dependently and -independently, respectively. This NO dependency can be applied to inflammatory reaction-related functional classifications, such as cell migration, chemotaxis, cytokine, Jak/STAT signaling pathway, and MAPK signaling pathway. Our results suggest that LPS-induced gene transcripts in inflammation or infection can be classified into physiological and toxic effects by their dependency on the NOS2-mediated NO release.

• Korean Journal of Food Science and Technology
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• v.49 no.3
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• pp.331-337
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• 2017

To evaluate the antioxidant activity of hexane, ethyl acetate, and butanol fractions obtained from Agastache rugosa extract, we measured the total polyphenol levels, DPPH radical scavenging activity, and reducing power. The ethyl acetate fraction of A. rugosa (AREA) displayed high phenolic levels, potent DPPH radical scavenging effect, and powerful reducing power. In addition, we examined the ability of AREA to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated BV-2 microglia. AREA suppressed NO production and inducible nitric oxide synthase (iNOS) expression and downregulated interleukin-6 (IL-6) mRNA level in LPS-stimulated BV-2 microglia. Furthermore, we detected rosmarinic acid in AREA by HPLC, which suggested that rosmarinic acid could be one of the bioactive materials responsible for the antioxidant and anti-inflammatory activities of AREA. These results suggested that AREA may be a good source of functional foods with antioxidant and anti-inflammatory activities.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.11
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• pp.1664-1674
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• 2016

This research analyzed the effect of ${\beta}$-glucan that is expected to alleviate the production of the inflammatory mediator in macrophagocytes, which are processed by the lipopolysaccharide (LPS) of Escherichia. The incubated layer was used for a nitric oxide (NO) analysis. The DNA-binding activation of the small unit of nuclear factor-${\kappa}B$ was measured using the enzyme-linked immunosorbent assay-based kit. In the RAW264.7 cells that were vitalized by Escherichia coli (E. coli) LPS, the ${\beta}$-glucan inhibited both the combatant and rendering phases of the inducible NO synthase (iNOS)-derived NO. ${\beta}$-Glucan increased the expression of the heme oxygenase-1 (HO-1) in the cells that were stimulated by E. coli LPS, and the HO-1 activation was inhibited by the tin protoporphyrin IX (SnPP). This shows that the NO production induced by LPS is related to the inhibition effect of ${\beta}$-glucan. The phosphorylation of c-Jun N-terminal kinases (JNK) and the p38 induced by the LPS were not influenced by the ${\beta}$-glucan, and the inhibitory ${\kappa}B-{\alpha}$ ($I{\kappa}B-{\alpha}$) decomposition was not influenced either. Instead, ${\beta}$-glucan remarkably inhibited the phosphorylation of the signal transducer and activator of transcription-1 (STAT1) that was induced by the E. coli LPS. Overall, the ${\beta}$-glucan inhibited the production of NO in macrophagocytes that was vitalized by the E. coli LPS through the HO-1 induction and the STAT1 pathways inhibition in this research. As the host immune response control by ${\beta}$-glucan weakens the progress of the inflammatory disease, ${\beta}$-glucan can be used as an effective immunomodulator.

• The Journal of Korean Medicine
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• v.22 no.3
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• pp.1-10
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• 2001

Objectives : The water extract of Nueihyuljunbang (NHJB) has long been used for treatment of ischemic brain damage in Oriental Medicine. However, little is known about the mechanism by which the water extract of NHJB recovers brain cens from ischemic damage. Methods : To elucidate the protective mechanism on ischemic induced cytotoxicity, we investigated the regulation of lipopolysaccharide (LPS) and phorbol-12-myristate-13-acetate (PMA)-induced inducible nitric oxide synthase (iNOS) expression in C6 glial cells. Results : LPS combined PMA treatment for 72 hours in C6 glial cells markedly induced nitric oxide (NO), but treatment of the cells with the water extract of NHJB decreased dose-dependently nitrite formation. In addition, LPS combined PMA treatment for 72 hours induced severe celt death and lactate dehydrogenase (LDH) release in C6 glial cells. However, treatment of the celts with the water extract of NHJB did not induce significant change compared to control cells. Furthermore, the protective effects of the water extract of NHJB were mimicked by the treatment of NGMMA, a specific inhibitor of NOS. LPS combined PMA induced iNOS activation in C6 glial cells caused chromosomal condensation and fragmentation of the nuclei by caspase activation. The treatment of C6 glial cells with the water extract of NHJB might suppress apoptosis via caspase inhibition by regulation of iNOS expression. Conclusions : From the results, we suggest that the protective effects of the water extract of NHJB against ischemic brain damage may be mediated by regulation of iNOS during ischemic condition.

• Molecules and Cells
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• v.28 no.4
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• pp.321-329
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• 2009

Rapid production of nitric oxide (NO) and reactive oxygen species (ROS) has been implicated in the regulation of innate immunity in plants. A potato calcium-dependent protein kinase (StCDPK5) activates an NADPH oxidase StRBOHA to D by direct phosphorylation of N-terminal regions, and heterologous expression of StCDPK5 and StRBOHs in Nicotiana benthamiana results in oxidative burst. The transgenic potato plants that carry a constitutively active StCDPK5 driven by a pathogen-inducible promoter of the potato showed high resistance to late blight pathogen Phytophthora infestans accompanied by HR-like cell death and $H_2O_2$ accumulation in the attacked cells. In contrast, these plants showed high susceptibility to early blight necrotrophic pathogen Alternaria solani, suggesting that oxidative burst confers high resistance to biotrophic pathogen, but high susceptibility to necrotrophic pathogen. NO and ROS synergistically function in defense responses. Two MAPK cascades, MEK2-SIPK and cytokinesis-related MEK1-NTF6, are involved in the induction of NbRBOHB gene in N. benthamiana. On the other hand, NO burst is regulated by the MEK2-SIPK cascade. Conditional activation of SIPK in potato plants induces oxidative and NO bursts, and confers resistance to both biotrophic and necrotrophic pathogens, indicating the plants may have obtained during evolution the signaling pathway which regulates both NO and ROS production to adapt to wide-spectrum pathogens.

• Journal of Applied Biological Chemistry
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• v.50 no.4
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• pp.264-269
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• 2007

Root extracts of Angelica gigas and A. acutiloba have been used traditionally for the treatment of gynecological diseases, as well as anemia, blood stasis, and inflammatory pain, as blood tonics in Oriental medicine. In the present study, we investigated the effects of A. gigas and A. acutiloba on inflammatory mediators in mouse macrophages and compared their activities. Many studies suggest that prostaglandin $E_2$ ($PGE_2$) biosynthesis and nitric oxide (NO) production play important roles in the processes of both inflammation and carcinogenesis. Ethanolic extracts from the roots of both species exhibited significant inhibitory effects on $PGE_2$ generation in lipopolysaccharide-stimulated RAW 264.7 cells. In particular, the extract from A. gigas was more effective than that from A. acutiloba. Although neither inhibited NO generation, the extract from A. acutiloba stimulated NO generation. Our results suggest that the roots of A. gigas might possess more anti-inflammatory and/or cancer chemopreventative activity than that of A. acutiloba due to the suppression of cyclooxygenase-2 (COX2)-mediated $PGE_2$ production. In addition, A. acutiloba might exert anti-tumor activity through an increase in macrophage-produced NO.

• Nutrition Research and Practice
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• v.11 no.2
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• pp.90-96
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• 2017

BACKGROUND/OBJECTIVES: Although the antioxidative effects of lycopene are generally known, the molecular mechanisms underlying the anti-inflammatory properties of lycopene are not fully elucidated. This study aimed to examine the role and mechanism of lycopene as an inhibitor of inflammation. METHODS/MATERIALS: Lipopolysaccharide (LPS)-stimulated SW 480 human colorectal cancer cells were treated with 0, 10, 20, and $30{\mu}M$ lycopene. The MTT assay was performed to determine the effects of lycopene on cell proliferation. Western blotting was performed to observe the expression of inflammation-related proteins, including nuclear factor-kappa B ($NF-{\kappa}B$), inhibitor kappa B ($I{\kappa}B$), mitogen-activated protein kinase (MAPK), extracellular signal-related kinase (ERK), c-jun NH2-terminal kinase (JNK), and p38 (p38 MAP kinase). Real-time polymerase chain reaction was performed to investigate the mRNA expression of tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), interleukin-1 beta ($IL-1{\beta}$), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Concentrations of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) were determined via enzyme-linked immunosorbent assays. RESULTS: In cells treated with lycopene and LPS, the mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, iNOS, and COX-2 were decreased significantly in a dose-dependent manner (P < 0.05). The concentrations of $PGE_2$ and NO decreased according to the lycopene concentration (P < 0.05). The protein expressions of $NF-{\kappa}B$ and JNK were decreased significantly according to lycopene concertation (P < 0.05). CONCLUSIONS: Lycopene restrains $NF-{\kappa}B$ and JNK activation, which causes inflammation, and suppresses the expression of $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, COX-2, and iNOS in SW480 human colorectal cancer cells.

• Korean Journal of Food Science and Technology
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• v.52 no.3
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• pp.249-254
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• 2020

Argyreia capitata (Vahl) Choisy is a species of the genus Argyreia. Although many studies have analyzed the biological activity of A. nervosa, A. speciasa, and A. acuta, the anti-inflammatory effect of A. capitata extract (ACE) remains unclear. In this study, we evaluated the anti-inflammatory effect of ACE using lipopolysaccharide (LPS)-induced inflammatory markers in RAW 264.7 cells. We confirmed that the ACE inhibited the LPS-induced NO (nitric oxide) and iNOS (inducible nitric oxide synthase) expression in RAW 264.7 cells. ACE suppressed not only the LPS-induced phosphorylation of IKK, IκB, and p65 but also IL-1β expression. Collectively, these results suggest that ACE is a novel anti-inflammatory agent that suppresses iNOS expression, NO production, and the NF-κB signaling pathways.