• IMMUNE NETWORK
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• v.14 no.6
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• pp.321-327
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• 2014

TGF-${\beta}$ induces IgA class switching by B cells. We previously reported that Smad3 and Smad4, pivotal TGF-${\beta}$ signal-transducing transcription factors, mediate germline (GL) ${\alpha}$ transcription induced by TGF-${\beta}1$, resulting in IgA switching by mouse B cells. Post-translational sumoylation of Smad3 and Smad4 regulates TGF-${\beta}$-induced transcriptional activation in certain cell types. In the present study, we investigated the effect of sumoylation on TGF-${\beta}1$-induced, Smad3/4-mediated $GL{\alpha}$ transcription and IgA switching by mouse B cell line, CH12F3-2A. Overexpression of small ubiquitin-like modifier (SUMO)-1, SUMO-2 or SUMO-3 did not affect TGF-${\beta}1$-induced, Smad3/4-mediated $GL{\alpha}$ promoter activity, expression of endogenous $GL{\alpha}$ transcripts, surface IgA expression, and IgA production. Next, we tested the effect of the E3 ligase PIASy on TGF-${\beta}1$-induced, Smad3/4-mediated $GL{\alpha}$ promoter activity. We found that PIASy overexpression suppresses the $GL{\alpha}$ promoter activity in cooperation with histone deacetylase 1. Taken together, these results suggest that SUMO itself does not affect regulation of $GL{\alpha}$ transcription and IgA switching induced by TGF-${\beta}1$/Smad3/4, while PIASy acts as a repressor.

• The Journal of Korean Obstetrics and Gynecology
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• v.34 no.4
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• pp.12-29
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• 2021

Objective: This study was performed to investigate the inhibitory effect of Cheongpochukeo-tang (CCT) on lipopolysaccharide (LPS)-induced inflammation model. Methods: RAW 264.7 cells were pre-treated with CCT and incubated with LPS (500 ng/ml) after 1 hour. Cell viability was measured by MTT assay to figure out cytotoxicity of CCT. The production of nitric oxide and mRNA expression of pro-inflammatory cytokine were measured. And the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) were examined to figure out molecular mechanisms of CCT's anti-inflammatory effects. In addition, mice survival rate and cytokine levels of serum were observed after treated with CCT. And mice liver tissues were observed and their cytokines levels in liver tissue were measured. Results: CCT did not have cytotoxic effect in RAW 264.7 cells. It inhibited LPS-induced nitric oxide (NO) production, but showed an increase in NO by itself at 2 mg/ml concentration. CCT inhibited mRNA expression of IL-1β, IL-6, TNF-α in a dose dependant and the activaton of MAPKs and NF-κB. In addition, CCT reduced mortality in the LPS-induced mouse model and inhibited production of cytokines in mouse serum and liver tissue. Conclusion: The results suggest that CCT could reduce LPS-induced inflammation by inhibiting MAPKs and NF-κB activaton, NO production, and pro-inflammatory cytokines secretion. Thereby, CCT could be effective medicine for the inflammatory disease.

• BMB Reports
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• v.45 no.2
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• pp.126-131
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• 2012

We have previously identified a DNA silent region located downstream of the 3'-end of the ${\beta}^{major}$ globin gene (designated B1-559) that contains a B1 retrotransposon, consensus binding sites for erythroid specific transcription factors and shares the capacity to act as promoter in hematopoietic cells interacting with ${\beta}$-globin gene LCR sequences in vitro. In this study, we have cloned four new non-polyA RNA transcripts being detected upon blockade of murine erythroleukemia (MEL) cell differentiation to erythroid maturation by methylation inhibitors and demonstrated that two of them share high structural homology with sequences of B1 element found within the B1-559 region. Although it is not clear yet whether and how these RNAs interfere with induction of erythroid maturation, these data provide evidence for the first time showing that methylation inhibitors can activate silent repetitive DNA sequences in MEL cells and may have implications in cancer chemotherapy using demethylating drugs as antineoplastic agents.

• IMMUNE NETWORK
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• v.10 no.4
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• pp.126-134
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• 2010

Background: $CD8^+$ T cells contribute to the clearance of Hepatitis B virus (HBV) infection and an insufficient $CD8^+$ T cell response may be one of the major factors leading to chronic HBV infection. Since the HBx antigen of HBV can up-regulate cellular expression of several immunomodulatory molecules, we hypothesized that HBx expression in hepatocytes might affect $CD8^+$ T cell activity. Methods: We analyzed the activation and apoptosis of $CD8^+$ T cells co-cultured with primary hepatocytes rendered capable of expressing HBx by recombinant baculovirus infection. Results: Expression of HBx in hepatocytes induced low production of $interferon-{\gamma}$ and apoptosis of CD8+ T cells, with no effect on CD8 T cell proliferation. However, transcriptional levels of H-2K, ICAM-1 and PD-1 ligand did not correlate with HBx expression in hepatocytes. Conclusion: Our results suggest that HBx may inhibit $CD8^+$ T cell response by regulation of $interferon-{\gamma}$ production and apoptosis.

• YAKHAK HOEJI
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• v.47 no.3
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• pp.190-194
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• 2003

As previously reported, an immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1, specific for a cell wall B-l,2-mannotriose, was protective against vaginal infection due to Candida albicans when mice were treated with the antibody. In this study, the role of neutrophil was examined in the protective effect of MAb B6.1 against vaginal infection. To deplete neutrophils, mice were given intravenously rat anti-mouse neutrophile MAb RB6-8C5 prior to intraperitoneal administration of MAb B6.1 to these mice. The mice were examined for antibody in their reproductive tract. By an ELISA, MAb B6.1 was found in the vaginal homogenates, but no antibody was detected in vaginal lavage materials. The neutropenia was induced by a single dose of the anti-neutrophil antibody, but lymphocytes were also partially depleted. The protective effect of MAb B6.1 was decreased when mice pretreated with MAb RB6-8C5 were given the anti-Candida antibody before challenge with C. albicans yeast cells intravaginally. These results show that neutrophils are involved in the MAb B6.1 protection against Candida vaginal infection.

• Korean Journal of Medicinal Crop Science
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• v.18 no.2
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• pp.105-112
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• 2010

Korean Sophora japonica has been found to posses an anti-inflammatory activity. In this study, Korean Sophora japonica extract, rutin, was used to know whether rutin inhibits to produce inflammatory mediators NO and TNF-$\alpha$ from the mouse peritoneal macrophages that were treated an inflammatory agent LPS. The rutin-1 hr pretreated macrophages were incubated with LPS for 0.5~5 hrs, and then collected the supernatant and the cell lysate for measurements of the level of iNOS, NO, TNF-$\alpha$ mRNA, TNF-$\alpha$, and p-NF-${\kappa}B$. Minimal and maximal effective doses of the rutin on them were 1 and $100{\mu}g/ml$, respectively. The maximal effective dose of rutin certainly inhibted the productions of iNOS, NO, TNF-$\alpha$ mRNA, TNF-$\alpha$and p-NF-${\kappa}B$ from the LPS-treated macrophages (p<0.0001). Its $ED_{50}$ for inhibition of TNF-$\alpha$ mRNA and p-NF-${\kappa}B$ was $5{\mu}g/m{\ell}$, and for iNOS, NO, and TNF-$\alpha$ was $10{\mu}g/m{\ell}$. The rutin did not have any cytotoxic effect. As the results, the Sophora japonica rutin could be a good candidate for an anti-inflammatory action.

• The Korea Journal of Herbology
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• v.28 no.6
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• pp.15-23
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• 2013

Objectives : To clarify the possible effects of Sinapis Semen and Raphani Semen on the development of pulmonary eosinophilic inflammation in a asthmatic mouse model. Methods : BALBav/c mice were sensitized to OVA followed intratracheally and by aerosol allergene challenges. We investigated the effect of Sinapis Semen and Raphani Semen on airway hyperresponsiveness, eosinophiic infitratio, immune cell phenotype, The2 cytokine product, and OVA-spedific IgE production. Results : Total lung cells, eosinophils, and lung leukocytes, OVA specific IgE levels, and Th 2cytokine levels such as IL-5, IL-13, IL-17, TNF-alpha, and eotaxin in BALF were reduced compared with those of OVA sensitized asthma mice (control). The absolute numbers of $CD3^+$, $CD3^+/CD69^+$, $CD3^-/CCR3^+$, $CD4^+$, $CD8^+$, $Gr-1^+/CD11b^+$, $B220^+/CD22^+$, $B220^+/IgE^+$ cells in lung tissiues significantly reduced compared to those of control. Specially total lung cells in BALF and the absolute number of $CD3^+/CD69^+$ and, $B220^+/IgE^+$ cells in lung tissiue effectively reduced in Sinapis Semen plus Raphani Semen compared to those of Sinapis Semen and Raphani Semen. Conclusions : These results indicate that Sinapis Semen plus Raphani Semen has deep inhibitory effects on airway inflammation and hyperresponsiveness in asmatic mouse model and also has effect of suppression of IL-5, IL-13, IL-17, OVA specific IgE production in BALF. The results verified that Sinapis Semen, Raphani Semen, and Sinapis Semen plus Raphani Semen could act as a immunomodulator which possess anti-inflammatory and anti-asthmatic property by modulating the relationship of Th1/Th2 cytokine imbalance.

• The Korea Journal of Herbology
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• v.33 no.4
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• pp.35-41
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• 2018

Objectives : Hyeonggaeyeongyotang Gagambang (HYT) is a herbal medicine prescribed for the treatment of inflammatory diseases, but it is necessary to study the exact therapeutic efficacy. This study aims to investigate the antibacterial and anti-inflmmatory activities of HYT. Methods : Antibacterial activity of HYT was confirmed by staining Escherichia coli, a gram negative strain, and Staphylococcus aureus, a gram positive strain, on solid Lysogeny Broth (LB) medium containing HYT. Antioxidant activity of HYT was confirmed by 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. The phosphorylation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha ($I{\kappa}B{\alpha}$) after lipopolysaccharide (LPS) treatment with HYT-treated RAW 264.7 mouse macrophages cells was confirmed by immunoblot analysis and the level of interleukin 1 beta (IL-$1{\beta}$) mRNA expression level was confirmed by quantitative real-time PCR. Results : HYT showed a concentration-dependent antibacterial activity against Escherichia coli and Staphylococcus aureus and also showed excellent antioxidant activity. HYT treatment attenuated the phosphorylation of $I{\kappa}B{\alpha}$induced by LPS treatment in RAW 264.7 mouse macrophages cells. The phosphorylation of $I{\kappa}B{\alpha}$is crucial for the regulation of the expression of various pro-inflammatory cytokines. In addition, IL-$1{\beta}$ mRNA expression level of RAW 264.7 mouse macrophages cells stimulated by LPS treatment was also inhibited by HYT treatment. Conclusions : Through experimental demonstration of the antioxidative, antimicrobial and anti-inflammatory effects of HYT, we demonstrated that HYT is a herbal medicine effective for the treatment of inflammatory diseases caused by various bacterial infections.

• Archives of Pharmacal Research
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• v.28 no.3
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• pp.274-279
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• 2005

Pharmacological inhibition of interleukin-12 (IL-12) production may be a therapeutic strategy for preventing the development and progression of disease in experimental models of autoimmunity. In this study, the effects of myricetin, a naturally occurring flavonoid present in fruits, vegetables and medicinal herbs, on the production of IL-12 were investigated in mouse macrophages stimulated with lipopolysaccharide (LPS). Myricetin significantly inhibited the LPS­induced IL-12 production from both primary macrophages and the RAW264.7 monocytic cell-line in a dose-dependent manner. The effect of myricetin on IL-12 gene promoter activation was analyzed by transfecting RAW264.7 cells with IL-12 gene promoter/luciferase constructs. The repressive effect was mapped to a region in the IL-12 gene promoter containing a binding site for NF-${\kappa}B$. Furthermore, activation of macrophages by LPS resulted in markedly enhanced binding activity to the NF-${\kappa}B$ site, which significantly decreased upon addition of myricetin, indicating that myricetin inhibited IL-12 production in LPS-activated macrophages via the down­regulation of NF-KB binding activity.

• Preventive Nutrition and Food Science
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• v.8 no.1
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• pp.96-104
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• 2003

Ginger (Zingiber officinale Roscoe) is traditionally used as appetite enhancer, improver of the digestive system, antithusive, anti-cold, antipyretic, analgesic, and antiinflammation. In vitro evaluation using human lymphocyte cultures showed almost similar indication with those in in vivo mouse study, NK cell lysing activity was improved significantly. Proliferation activity of B and T cells, and CD3$^{＋}$ and CD3$^{＋}$CD4$^{＋}$T cell subset were better observed using oleoresin or gingerol and shogaol fractions. Although there were higher activities in gingerol, the improvement was almost equal to that by oleoresin. Shogaol did not show better improvement except at higher concentration. It could be concluded that treatment with single bioactive compound, such as gingerol, did not show significant effects compared to oleoresin, the crude extract. In human study, involving healthy male adult, the improvement of NK cell lysing activity was again demonstrated and even more apparent. The mechanism involved in the protection seemed to be through the antioxidant activity of gingerol. However, other mechanism underlying the improvement of NK cell lysing activity must be involved since this improvement seemed to be specifically toward NK cell activity. Since NK cells ave specific for the elimination of virus-infected cell and mutated cells, this positive effect on the immune system are very interesting. This work has also scientifically proved that the traditional beliefs that ginger had preventive effects on common cold appeared to be reasonable.