• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.1
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• pp.39-45
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• 2010

Chlorella has been reported to have certain beneficial physiological effects, including hypocholesterolemic, antihypertensive, antioxidative, and anti-tumor activities in animal and human studies. The aim of this study was to determine the anti-inflammatory activities of an 80% methanol extract(CE-Met), hexane fraction (CE-Hex), and ethyl acetate fraction (CE-EA) of Chlorella ellipsoidea in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage. Treatment with various concentrations of the C. ellipsoidea extract resulted m a significant, dose-dependent reduction in nitric oxide (NO) production by LPS-induced macrophages. The C. ellipsoidea extract significantly inhibited LPS-induced NO production accompanied by an attenuation of IL-6 and TNF-$\alpha$ formation in macrophages. These results suggest potent inhibit이y effects on the production of inflammatory mediators by a C. ellipsoidea extract. Thus, C. ellipsoidea extract may be a potent anti-inflammatory agent for troubled skin.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.98-105
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• 1999

Intestinal bacteria comprise one-third of the contents of the large intestine in humans. Their interactions with the gastrointestinal immune system induce characteristic immunological responses which stimulate or suppress the host's defense system. RAW 264.7 murine cell line was used as a macrophage model to assess the effects of the exposure to the isolated human intestinal bacteria, Bacteroides, Bifidobacterium, Eubacterium, Streptococcus, and E. coli, on NO (nitric oxide), $H_2O_2$(hydrogen peroxide), and cytokines IL (interleukin)-6 and TNF (tumor necrosis factor)-a production. RAW 264.7 cells were cultured in the presence of heat-killed bacteria for 24 h at concentrations of 0-$50\mu$g/ml. Our results showed that Bacteroides and E. coli stimulated IL-6, TNF-$\alpha$, NO, and $H_2O_2$production at high levels even at $1\mu$g/ml, whereas Bifidobacterium, Eubacterium, and Streptococcus showed a low level of stimulation at $1\mu$g/ml, and a gradual increase as the cell concentration increased up to $50\mu$g/ml. This result suggests that gram-negative Bacteroides and E. coli are better able to stimulate macrophage than gram-positive Bifidobacterium, Streptococcus, and Eubacterium. The in vitro approaches employed here should be useful in further characterization of the effects of intestinal bacteria on gastrointestinal and systemic immunity.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1333-1340
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• 2016

The main objective of this study was to investigate whether Lactobacillus rhamnosus GG (LGG) ameliorated the effects of Citrobactor rodentium infection in Toll-like receptor 2 (TLR2) knockout (KO) and TLR4 KO mice, as well as in wild-type C57BL/6 (B6) mice. TLR2 KO, TLR4 KO, and B6 mice were divided into three groups per each strain. Each group had an uninfected control group (n = 5), C. rodentium-infected group (n = 8), and LGG-pretreated C. rodentium-infected group (n = 8). The survival rate of B6 mice infected with C. rodentium was higher when pretreated with LGG. Pretreatment with LGG ameliorated C. rodentium-induced mucosal hyperplasia in B6 and TLR4 KO mice. However, in C-rodentium-infected TLR2 KO mice, mucosal hyperplasia persisted, regardless of pretreatment with LGG. In addition, LGG-pretreated B6 and TLR4 KO mice showed a decrease in spleen weight and downregulation of tumor necrosis factor alpha, interferon gamma, and monocyte chemotactic protein 1 mRNA expression compared with the non-pretreated group. In contrast, such changes were not observed in TLR2 KO mice, regardless of pretreatment with LGG. From the above results, we conclude that pretreatment with LGG ameliorates C. rodentium-induced colitis in B6 and TLR4 KO mice, but not in TLR2 KO mice. Therefore, LGG protects mice from C. rodentium-induced colitis in a TLR2-dependent manner.

• YAKHAK HOEJI
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• v.52 no.1
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• pp.73-78
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• 2008

Immune system can protect host attacking from a variety of microorganism and virus through innate and adaptive immunities. The innate immune system can be activated by recognition of conserved carbohydrates on the cell surface of pathogen resulting in protection, immunity regulation and inflammation. Immunostimulating and anti-tumor ${\beta}$-glucan, major cell wall component of many fungi, could be recognized as pathogen associated molecular pattern (PAMP) by C-type lectin such as pathogen recognition receptor (PRR) of host innate immunity cells. In spite of many studies of basidiomycetes ${\beta}$-glucan on immunostimulation, little is known about the precise mechanism as molecular-level. Among C-type lectins, dectin-1 was cloned and reported as a ${\beta}$-glucan receptor. In this report, we demonstrated induction of cytokine gene transcription by Ganoderma lucidum ${\beta}$-glucan in the absence or presence of lipopolysaccharide (LPS) by RT-PCR analysis. The expression of murine dectin-1 (MD-1) on RAW264.7 macrophage by RT-PCR showing both the full length, 757 bp $(MD-1{\alpha})$ and alternative spliced form, 620 bp $(MD-1{\beta})$. Both $MD-1{\alpha}$ and $MD-1{\beta}$ mRNAs were induced by ${\beta $-glucan both in the absence and presence of LPS. To explore expression of inflammatory cytokines by ${\beta}$-glucan, RAW264.7 cells were treated with ${\beta}$-glucan for 12 hours. As a result, the expressions of IL-1 IL-6, IL-l0 and $TNF-{\alpha}$ were increased by ${\beta}$-glucan treatment in a dose-dependent fashion. From these results, ${\beta}$-glucan induced transcriptions of dectin-1 and immune activating cytokine genes, indicating induction of immune allertness by expressing dectin-1 and secreting inflammatory cytokines.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.295-301
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• 2003

Background: Carcinoembryonic antigen (CEA) is well-known soluble tumor marker frequently detectable in peripheral blood of carcinoma patients and considered as good target for antigen-specific immunotherapy. In this study, we used a replication-deficient adenovirus containing CEA to study CTL induction in vitro after adenovirus-mediated gene transfer into DC. Methods: DC were obtained from mouse bone marrow and cultured with IL-4 and GM-CSF. For measuring CTL activity, splenocytes were harvested from the mice, which were immunized with DC that had been infected AdV-CEA or pulsed with CEA peptide. Untreated DC was used as a control. Splenocytes were re-stimulated in vitro with DC pulsed with CEA peptide for 7 days and CTL activity with CEA peptide-pulsed EL-4 cells were assessed in a standard $^{51}Cr$-release assay. The frequencies of antigen-specific cytokine-secreting T cell were determined with $mIFN-{\gamma}$ELISPOT. Results: DC infected with recombinant adenovirus expressing CEA induced CEA-specific CTL responses in vivo. Splenocyte induced from mice immunized with AdV-CEA-infected DC increase in the number of $IFN-{\gamma}$ secreting T cells compared with those from mice immunized with CEA peptide-pulsed DC. Conclusion: These results suggested that DC infected with recombinant adenovirus has advantages over other forms of vaccination and could provide an alternative approach vaccination therapies.

• The Korea Journal of Herbology
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• v.27 no.3
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• pp.31-38
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• 2012

Objectives : Mori Folium is one of the traditional medicinal herb. It was commonly used for sericulture in the world and has been traditionally administered as natural therapeutic agent for the treatment of filariasis, diabetes and dropsy in East Asia. This study investigated an anti-inflammatory potential of Mori Folium ethanol extract (MFE). Methods : We examined the effects of MFE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in a murine macrophage cell line, RAW 264.7. Results : MFE inhibited production of NO and $PGE_2$ in a dose dependent manner and also decreased the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2, interleukin (IL)-1, IL-6 and tumor necrosis factor-${\alpha}$. As a plausible molecular mechanism, increased degradation of I-${\kappa}B{\alpha}$ and phosphorylation of I-${\kappa}B{\alpha}$, NF-${\kappa}B$ and MAP kinases by LPS were partly blocked by MFE treatment. Conclusions : These results suggest that MFE has an anti-inflammatory therapeutic potential, which may result from inhibition of NF-${\kappa}B$ activation and MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

• BMB Reports
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• v.51 no.10
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• pp.514-519
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• 2018

Ovarian cancer is the most fatal gynecological malignancy in women and identification of new therapeutic targets is essential for the continued development of therapy for ovarian cancer. TRRAP (transformation/transcription domain-associated protein) is an adaptor protein and a component of histone acetyltransferase complex. The present study was undertaken to investigate the roles played by TRRAP in the proliferation and tumorigenicity of ovarian cancer stem cells. TRRAP expression was found to be up-regulated in the sphere cultures of A2780 ovarian cancer cells. Knockdown of TRRAP significantly decreased cell proliferation and the number of A2780 spheroids. In addition, TRRAP knockdown induced cell cycle arrest and increased apoptotic percentages of A2780 sphere cells. Notably, the mRNA levels of stemness-associated markers, that is, OCT4, SOX2, and NANOG, were suppressed in TRRAP-silenced A2780 sphere cells. In addition, TRRAP overexpression increased the mRNA level of NANOG and the transcriptional activity of NANOG promoter in these cells. Furthermore, TRRAP knockdown significantly reduced tumor growth in a murine xenograft transplantation model. Taken together, the findings of the present study suggest that TRRAP plays an important role in the regulation of the proliferation and stemness of ovarian cancer stem cells.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.975-979
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• 2006

The extracts of Oenanthe javanica were evaluated for their effects on the expression of cyclooxygenase-2 (COX-2), which is mediated by the translocation of the p65 subunit into the nucleus. Fractions of ethyl acetate and chloroform from 80% ethanol extracts of O. javanica exhibited inhibitory effects on the secretion of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) from lipopolysaccharide (LPS)-stimulated peritoneal macrophages; however, the aqueous- and hexane-fractions showed no significant effect. The ethyl acetate- and chloroform-fractions also reduced the COX-2 enzyme levels after 24-hr treatment. RT-PCR showed that the mRNA levels of COX-2 decreased following treatment with these fractions, suggesting that COX-2 expression is transcriptionally regulated by these extracts. We examined the effects of the chloroform- and ethyl acetate-fractions on the cytosolic activation of nuclear factor-${\kappa}B$ ($NF-{\kappa}B$, p65 subunit) and on the degradation of inhibitor-${\kappa}B{\alpha}$ ($I-{\kappa}B{\alpha}$) in order to determine the mechanism of COX-2 regulation. The LPS-stimulated activation of the p65 subunit was significantly blocked upon the addition of $50\;{\mu}g/mL$ of these fractions, and the cytosolic $I-{\kappa}B{\alpha}$ degradation process was simultaneously inhibited. These findings suggest that the inhibition of COX-2 expression by the ethyl acetate-and chloroform-fractions may result from the inhibition of p65 translocation by blocking the degradation of $I-{\kappa}B{\alpha}$; this may be the mechanistic basis for the anti-inflammatory effects of O. javanica.

• Journal of Ginseng Research
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• v.36 no.3
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• pp.263-269
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• 2012

Korean red ginseng has shown therapeutic effects for a number of disease conditions. However, little is known about the anti-inflammatory effect of Korean red ginseng saponin fraction (RGSF) in vitro and in vivo. Therefore, in this study, we showed that RGSF containing 20(S)-protopanaxadiol type saponins inhibited nitric oxide production and attenuated the release of tumor necrotic factor (TNF)-${\alpha}$, interleukin (IL)-6, granulocyte monocyte colony stimulating factor (GMCSF), and macrophage chemo-attractant protein-1 in lipopolysaccharide (LPS) stimulated murine macrophage RAW264.7 cells. Moreover, RGSF down-regulated the mRNA expressions of inducible nitric oxide synthase, cyclooxyginase-2, IL-$1{\beta}$, TNF-${\alpha}$, GMCSF, and IL-6. Furthermore, RGSF reduced the level of TNF-${\alpha}$ in the serum and protected mice against LPS mediated endotoxic shock. In conclusion, these results indicated that ginsenosides from RGSF and their metabolites could be potential sources of therapeutic agents against inflammation.

• Journal of Ginseng Research
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• v.32 no.4
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• pp.315-323
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• 2008

In the previous studies, we isolated the compound K rich fractions (CKRF) and showed that CKRF inhibited Toll-like receptor (TLR) 4- or TLR9-induced inflammatory signaling. To extend our previous studies,1) we investigated the molecular mechanisms of CKRF in the TLR4-associated signaling via nuclear factor (NF)-${\kappa}B$, and in vivo role of CKRF for induction of tolerance in lipopolysaccharide (LPS)-induced septic shock. In murine bone marrow-dervied macrophages, CKRF significantly inhibited the induction of mRNA expression of proinflammatory mediators such as tumor necrosis factor-${\alpha}$, interleukin-6, cyclooxygenase-2, and inducible nitric oxide synthase. In addition, CKRF significantly attenuated the transcriptional activities of TLR4/LPS-induced NF-${\kappa}B$. Nuclear translocation of NF-${\kappa}B$ in response to LPS stimulation was significantly abrogated by pre-treatment with CKRF. Furthermore, CKRF inhibited the recruitment of p65 to the interferon-sensitive response element flanking region in response to LPS. Finally, oral administration of CKRF significantly protected mice from Gram-negative bacterial LPS-induced lethal shock and inhibited systemic inflammatory cytokine levels. Together, these results demonstrate that CKRF modulates the TLR4-dependent NF-${\kappa}B$ activation, and suggest a therapeutic role for Gram-negative septic shock.