• Journal of Pharmacopuncture
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• v.14 no.2
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• pp.15-24
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• 2011

Background: Type I allergy is involved in allergic asthma, allergic rhinitis, and atopic dermatitis which are accompanied by an acute and chronic allergic inflammatory responses. Rehmannia glutinosa is a traditional medicine in the East Asian region. This study examined whether a Rehmannia Glutinosa pharmacopuncture solution (RGPS) had anti-allergic or anti-inflammatory effects in antigen-stimulated-RBL-2H3 cells. Methods: We determined the effect of RGPS on cell viability using the 3-[4,5-dimethylthiazolyl]-2,5-diphenyltetrazolium bromide (MTT) assay. We also examined the effect of RGPS on the release of ${\beta}$-hexosaminidase and the secretion of IL-4 and TNF-${\alpha}$ using ELISA. In addition, we evaluated the effect of RGPS on the mRNA expression of various cytokines; IL-2, IL-3, IL-4, IL-5, IL-13 and TNF-${\alpha}$ using RT-PCR. Furthermore, we assessed the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-${\kappa}$B using Western blotting after RGPS treatment. Results: We found that RGPS ($10^{-4}$ to $10^{-1}$ dilution) did not cause any cytotoxicity. We observed significant inhibition of ${\beta}$-hexosaminidase release and suppression of the protein secretion of IL-4 and TNF-${\alpha}$ and mRNA expression of multiple cytokines in antigen-stimulated-RBL-2H3 cells after RGPS treatment. Additionally, RGPS suppressed not only the phosphorylation of MAPKs, but also the transcriptional activation of NF-${\kappa}$B in antigen-stimulated-RBL-2H3 cells. Conclusions: These results suggest that RGPS inhibits degranulation and expression of cytokines including IL-4 and TNF-${\alpha}$ via down-regulation of MAPKs and NF-${\kappa}$B activation in antigen-stimulated-RBL-2H3 cells. In conclusion, RGPS may have beneficial effects in the exerting anti-allergic or anti-inflammatory activities.

• Biomolecules & Therapeutics
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• v.13 no.3
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• pp.190-197
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• 2005

Advanced glycation end products (AGE) have been implicated in the pathogenesis of diabetic complications including nephropathy. However, the role of AGE in the activation of mesangial cells cultured under high glucose has not been elucidated. The effects of aminoguanidine, which prevents formation of AGE and protein cross-linking, on the synthesis of $TGF-{\beta}1$ and fibronectin by rat mesangial cells cultured under high glucose for 2 weeks were examined and compared with the effects of $N^G$-nitro-L-arginine methyl ester (NAME), a selective nitric oxide synthase inhibitor, because aminoguanidine also inhibits the inducible nitric oxide synthase. Culture of mesangial cells in 30 mM (high) glucose for 2 weeks induced 1.5-fold (ELISA) and 1.9-fold (Western blot analysis) increase in AGE in the culture media compared to 5.6 mM (control) glucose. Northern blot analysis revealed 1.5-fold increase in $TGF-{\beta}1$ and 1.7-fold increase in fibronectin mRNA expression in cells cultured under high glucose compared to control glucose. Increases in mRNA expression were followed by increased protein synthesis. Mink lung epithelial cell growth inhibition assay revealed 1.4-fold increase in $TGF-{\beta}1$ protein in high glucose media compared to control. Fibronectin protein also increased 2.1-fold that of control glucose by Western blot analysis. Administration of aminoguanidine suppressed AGE formation in a dose dependent manner and at the same time suppressed $TGF-{\beta}1$ and fibronectin synthesis by mesangial cells cultured in both control and high glucose. In contrast, NAME did not affect high glucose-induced changes. These findings support a role for AGE in high glucose-induced upregulation of $TGF-{\beta}1$ and fibronectin synthesis by mesangial cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.6
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• pp.1832-1842
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• 2004

The present study was performed to examine the possible anti-inflammatory effects of a herbal drug ASCH in RAW264.7 cell line. Inflammation was induced by LPS toxin treatment to RAW264.7 macrophage cell line. Increases in cytokine production such as IL-1β, IL-6. and IL-18, COX-2, NOS-Ⅱ (iNOS), and TNF-alpha were observed at mRNA level in the LPS-treated RAW264.7 cells. Measurement of IL-6, nitric oxide and the reactive oxygen species (ROS) showed increased production of these inflammation mediators. Treatment of ASCH effectively decreased IL-1β protein in a dose-dependent manner, and IL-6 and IL-18 were reduced at 100㎍/㎖ of ASCH concentration. NO production was also decreased by ASCH treatment. A slight inhibition for TNF-alpha in terms of protein, but not mRNA level was obtained by 100㎍/㎖ of ASCH treatment. ASCH treatment to normal RAW264.7 cells did not produce any cytotoxicity, indicting that the action of ASCH was selective to inflammatory cells. Thus, the present data suggest that ASCH may act as an important regulator to alleviate the inflammatory symptoms.

• The Journal of Pediatrics of Korean Medicine
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• v.26 no.1
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• pp.36-45
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• 2012

Objectives Perillae Folium (PF) has been widely used in Korean herbal medicines used for treatment of acute and chronic inflammatory diseases, such as rhinitis, asthma, and enteritis. In this study, to investigate the protective effect of PF on type 1 allergic response, we determined whether PF inhibits early or late allergic responses. Methods The effect of PF was analyzed by ELISA,. RT-PCR and Western blot in RBL-2H3 cells. Levels of ${\beta}$-hexosaminidase, interleukin (IL)-4 and TNF-${\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISAs). mRNA levels of cytokines and enzymes were analyzed with RT-PCR. Signal transduction was analyzed with Western blot. Results We found that PF suppressed ${\beta}$-hexosaminidase release in RBL-2H3 by the IgE-DNP-HSA stimulation. PF also significantly inhibited enzymes level, such as COX-1, COX-2, iNOS, and HDC2, along with reduced cytokine levels, such as IL-2, IL-3, IL-4, IL-6, IL-13, and TNF-${\alpha}$ in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, and $I{\kappa}B{\alpha}$. Conclusions Our results indicate that PF protects against type 1 allergic response and exert an anti-inflammatory effect through the inhibition of degranulation and expression of cytokines and enzymes via the suppression of signal transduction.

• The Journal of Pediatrics of Korean Medicine
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• v.26 no.1
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• pp.1-15
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• 2012

$CheongGiGeoYangTang$ has been used for anti-allergenic purpose. However there was no experimental study about its effect. Therefore, this study was designed to investigate the anti-allergenic effect of $CheongGiGeoYangTang$. Methods Modifiability of RBL-2H3 mast cells' IL-4, IL-13 was analyzed by qRT-PCR and ELISA. Also, the suppressive effect of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors was observed by western blotting. OVA-specific IgE, IL-4 and IL-13 production in ovalbumin allergy model was examined as well. Results It was showed that the RBL-2H3 mast cells treated with $CheongGiGeoYangTang$ extract(CGGYT) was significantly suppressed mRNA expression, production of IL-4 and IL-13, and prominently inhibited the expression of transcription factors including GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 but not c-Jun. The administration of CGGYT was suppressed the amount of OVA-specific IgE, IL-4 and IL-13 in OVA/alum-sensitized mice. Conclusions We considered CGGYT would regulate the allergic inflammation as inhibition of IL-4 and IL-13 production in activated mast cells and Th2 cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.4
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• pp.768-773
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• 2002

Behcets disease is a systemic inflammatory disorder. The etiology and pathogenesis of Behcets disease has yet been fully elucidated but might involve immune dysfunction. Cytokines involved in the regulation of inflammatory reactions and immune responses may play a role in the pathogenesis of Behcets disease (BD). Onchungeum is an Oriental herbal medication, which has been successfully used in Korea for the treatment of BD. This report describes modulation effects of Onchungeum on cytokine production from phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) of Behcets patients by ELISA. Onchungeum significantly inhibited the production of pro-inflammatory cytokines. TNF-α and IL-1β, compared to absence of Onchungeum (by 52.3 1.4 % inhibition for TNF-α and 113.5 3.3 % for IL-1β, p < 0.001). Onchungeum also inhibited the production of IFN-γ, immunoregulatory Th1 cytokine, by 89.4 0.8 % (p < 0.001). The inhibitory effects of Onchungeum on cytokine production showed dose-dependent manner, and the pre-treatment of 1 mg/ml Onchungeum had better effects than immunosuppressive drug for treatment of BD, cyclosporin A. Our results suggest that Onchungeum treatment for Behcets disease patients may have pharmacologic activities and abilities of regulation of immune and inflammatory responses by cytokine modulation.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7719-7724
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• 2013

Background: Overexpression of survivin, a known inhibitor of apoptosis, is associated with tumor progression and drug resistance in numerous malignancies, including leukemias. The aim of this study was to investigate the effect of a specific survivin small interference RNA (siRNA) on proliferation and the sensitivity of HL-60 acute myeloid leukemia (AML) cells to the chemotherapeutic drug etoposide. Materials and Methods: The cells were transfected with siRNAs using Lipofectamine $^{TM}2000$ transfection reagent. Relative survivin mRNA and protein levels were measured by quantitative real-time PCR and Western blotting, respectively. Trypan blue exclusion assays were performed to monitor tumor cell proliferation after siRNA transfection. The cytotoxic effects of etoposide and survivin siRNA, alone and in combination, on leukemic cells were determined using MTT assay. Apoptosis was assessed by ELISA cell death assay. Results: Survivin siRNA markedly reduced both mRNA and protein expression levels in a time-dependent manner, leading to distinct inhibition of cell proliferation and increased spontaneous apoptosis. Surprisingly, survivin siRNA synergistically increased the cell toxic effects of etoposide. Moreover, survivin down-regulation significantly enhanced its induction of apoptosis. Conclusions: Our study suggests that down-regulation of survivin by siRNA can trigger apoptosis and overcome drug resistance of leukemia cells. Therefore, survivin siRNA may be an effective adjuvant in AML chemotherapy.

• The Journal of Korean Medicine
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• v.29 no.4
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• pp.146-160
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• 2008

Objectives: The present study was designated to evaluate the direct effects of Fructus Liquidambaris on suppression of eosinophil activity and on suppression of chemokines such as eotaxin, IL-8, ICAM-1, and VCAM-1, in vitro. Methods: A549 cells were stimulated with TNF-$\alpha$ (100 ng/ml), IL-4 (100 ng/ml) or IL-$1{\beta}$(10 ng/ml) to induce chemokines and adhesion molecules involved in eosinophil chemotaxis. Then after treatment of Fructus Liquidambaris, inhibition effect assay such as ELISA, real-time RT-PCR, and chemotaxis assay was performed. Results: Eotaxin level was suppressed in both protein secretion and mRNA expression. Eosinophil recruitment to lung epithelial cells was also reduced by Fructus Liquidambaris, implying the role of eotaxin in eosinophil recruitment. In addition, expression of IL-8 was also suppressed by Fructus Liquidambaris (p<0.05). However, expression of adhesion molecules (ICAM-1, VCAM-1) related to eosinophil was not affected. The eosinophil migration was inhibited at high concentration of Fructus Liquidambaris (p<0.05). Conclusion: These results suggest that Fructus Liquidambaris may regulate a common signaling pathway of eotaxin and IL-8. FS might be of therapeutic value in diseases such as asthma.

• Journal of Pharmacopuncture
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• v.11 no.1
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• pp.119-125
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• 2008

Objective : This experimental study was performed to investigate the effects of Houttuyniae Herba extract on anti-inflammation and anti-oxidation. Methods : The cytotoxicity of Houttuyniae Herba water extract and ethanol extract about viability of Raw 264.7 cell were tested using a colormetric tetrazolium assay(MTT assay). We investigated the inhibitory effects of Houttuyniae Herba water extract and ethanol extract on Propionibactrium acnes using paper disk diffusion method. To investigate the anti-inflammation effects of Houttuyniae Herba water extract and ethanol extract on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit. We evaluated anti-oxidation effects of Houttuyniae Herba water extract and ethanol extract on HaCaT cell by Enzyme recycling method. Results : 1. In Houttuyniae Herba water extract and ethanol extract, cell toxicity depended on the density and wasn't difference between two extracts. 2. Houttuyniae Herba water extract and ethanol extract has not the significant inhibition effect of Propionibactrium acnes. 3. Concentration of 50, $100{\mu}g/m{\ell}$ Houttuyniae Herba water extract inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 4. All extracts except for $20{\mu}g/m{\ell}$ Houttuyniae Herba water extract showed anti-oxidation effect by decreasing the DPPH radicals. Conclusion : These results indicate that Houttuyniae Herba extract has anti-inflammation and anti-oxidation effects. If further study is performed, the use of Houttuyniae Herba extract will be valuable and benificial in the therapy of Propionibactrium acnes.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.280-287
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• 2015

Current influenza vaccines are produced in embryonated chicken eggs. However, egg-based vaccines have various problems. To address these problems, recombinant protein vaccines have been developed as new vaccine candidates. Unfortunately, recombinant proteins frequently encounter aggregation and low stability during their biogenesis. It has been previously demonstrated that recombinantly expressed proteins can be greatly stabilized with high solubility by fusing stabilizing peptide (SP) derived from the C-terminal acidic tail of human synuclein (ATS). To investigate whether SP fusion proteins can induce protective immunity in mice, we produced influenza HA and SP fusion protein using a baculovirus expression system. In in vitro tests, SP-fused recombinant HA1 (SP-rHA1) was shown to be more stable than recombinant HA1 (rHA1). Mice were immunized intramuscularly with baculovirus-expressed rHA1 protein or SP-rHA1 protein ($2{\mu}g/mouse$) formulated with aluminum hydroxide. Antibody responses were determined by ELISA and hemagglutination inhibition assay. We observed that SP-rHA1 immunization elicited HA-specific antibody responses that were comparable to rHA1 immunization. These results indicate that fusion of SP to rHA1 does not negatively affect the immunogenicity of the vaccine candidate. Therefore, it is possible to apply SP fusion technology to develop stable recombinant protein vaccines with high solubility.