• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.52-66
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• 2001

Background : NF-${\kappa}B$ is the most important transcriptional factor in IL-8 gene expression. Triptolide is a new compound that recently has been shown to inhibit NF-${\kappa}B$ activation. The purpose of this study is to investigate how triptolide inhibits NF-${\kappa}B$-dependent IL-8 gene transcription in lung epithelial cells and to pilot the potential for the clinical application of triptolide in inflammatory lung diseases. Methods : A549 cells were used and triptolide was provided from Pharmagenesis Company (Palo Alto, CA). In order to examine NF-${\kappa}B$-dependent IL-8 transcriptional activity, we established stable A549 IL-8-NF-${\kappa}B$-luc. cells and performed luciferase assays. IL-8 gene expression was measured by RT-PCR and ELISA. A Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation and an electromobility shift assay was done to analyze NF-${\kappa}B$ DNA binding. p65 specific transactivation was analyzed by a cotransfection study using a Gal4-p65 fusion protein expression system. To investigate the involvement of transcriptional coactivators, we perfomed a transfection study with CBP and SRC-1 expression vectors. Results : We observed that triptolide significantly suppresses NF-${\kappa}B$-dependent IL-8 transcriptional activity induced by IL-$1{\beta}$ and PMA. RT-PCR showed that triptolide represses both IL-$1{\beta}$ and PMA-induced IL-8 mRNA expression and ELISA confirmed this triptolide-mediated IL-8 suppression at the protein level. However, triptolide did not affect $I{\kappa}B{\alpha}$ degradation and NF-$_{\kappa}B$ DNA binding. In a p65-specific transactivation study, triptolide significantly suppressed Gal4-p65T Al and Gal4-p65T A2 activity suggesting that triptolide inhibits NF-${\kappa}B$ activation by inhibiting p65 transactivation. However, this triptolide-mediated inhibition of p65 transactivation was not rescued by the overexpression of CBP or SRC-1, thereby excluding the role of transcriptional coactivators. Conclusions : Triptolide is a new compound that inhibits NF-${\kappa}B$-dependent IL-8 transcriptional activation by inhibiting p65 transactivation, but not by an $I{\kappa}B{\alpha}$-dependent mechanism. This suggests that triptolide may have a therapeutic potential for inflammatory lung diseases.

• International Journal of Clinical Preventive Dentistry
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• v.14 no.4
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• pp.264-270
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• 2018

Objective: This study performed a quantitative analysis using the real-time polymerase chain reaction technique to examine the oral microbial prevalence in adults and intended to examine the correlations between risk factors of periodontal disease and oral bacteria and correlation between oral test scores and oral microorganisms. Methods: We examined papillary marginal attached (PMA) index, modified patient hygiene performance (M-PHP) index, probing depth (PD), modified gingival index, and oral bacteria counts and surveyed 117, 20 years or older adult males and females who visited dental clinics in the Daejeon region to analyze the prevalence and oral health. Results: The prevalence was 100% for Fusobacterium nucleatum, meaning it was observed in all examined subject, 85.5% for Parvimonas micra, 76.1% for Prevotella intermedia, and 72.6% for Tannerella forsythia. The averages of P. gingivalis and T. forsythia increased as the examined subjects were older, and there was a statistically significant difference between T. forsythia and E. nodatum in relation to medical history, between P. intermedia and P. micra in relation to gender, and between P. intermedia and E. corrodens in relation to smoking (p<0.05). For a correlation between the oral test scores and oral microorganisms, P. gingivalis and F. nucleatum was highly correlated with PD (correlation coefficient of 0.51 and 0.41) (p<0.01) while P. gingivalis, P. micra, C. rectus, and E. nodatum were significantly correlated with M-PHP index, gingival index, PD, and PMA index (p<0.01, p<0.05). Conclusion: For oral health management of adults, the age, systemic disease, and smoking are closely related to oral bacteria, and P. gingivalis, T. forsythia, F. nucleatum, P. intermedia, P. micra, C. rectus, E. corrodens, and E. nodatum are considered to be the oral microorganisms that indicate periodontal health.

• Journal of Korean Medicine Rehabilitation
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• v.15 no.1
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• pp.25-37
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• 2005

목적 : 진피(Citus Unshiu, CU)는 실험적으로 항산화, 항알러지, 항종양 효과 등이 있다고 보고되고 있다. 본 연구에서는 비만세포에서 PMA와 A23187에 의하여 유도된 싸이토카인의 증가에 대한 진피 추출물의 억제 효과와 그 기전을 알아보고자 한다. 방법 : PMA+A23187에 의한 싸이토카인 생성에 대한 진피 추출물의 억제효과 기전을 조사하기 위하여 진피를 처리한 후 $IL-1{\beta}$, IL-8, $TNF-{\alpha}$의 생산 및 혈관내피성장인자 (VEGF), 과립구 대식구 군집 자극 인자 (GM-CSF), 저산소 유도 인자(HIF-1)의 발현을 조사하였다. 결과 : 실험에서 PMA와 A23187을 처리한 경우 대조군에 비하여 $IL-1{\beta}$, IL-8, $TNF-{\alpha}$의 생산을 유의하게 증가시켰으나 진피 추출물을 처리한 군에서는 유의하게 억제되었다. 특히 $IL-1{\beta}$의 생성은 $103.7{\pm}5%$, IL-8 생성은 $34.6{\pm}7%$, $TNF-{\alpha}$의 생성은 $85.9{\pm}4.5%$ 억제되었다. (P<0.05). 또한 진피 추출물은 PMA와 A23187에 의하여 증가한 VEGF, GM-CSF, $HIF-1{\alpha}$의 발현 증가를 억제하였다 (약 90.9%의 VEGF, 61.6%의 GM-CSF). 결론 : 이러한 결과는 진피가 염증반응에 대한 HIF-1의 억제자로 작용할 수 있으면, 진피가 비만세포 매개성 염증 질환 치료제의 후보 물질이 될 것으로 사료된다.

• The Korea Journal of Herbology
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• v.23 no.4
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• pp.81-89
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• 2008

Objectives: In this study, the pharmacological effects of the ethylacetate extract of Ostericum koreanum(North Kangwhal; NK) on allergic inflammation were investigated in activated human mast cells. Methods: North Kangwhal was extracted with 80% methanol for 24 h, and then fractionated with ethylacetate(NK-EtOAc extract). HMC-1 cells, an human mast line, were pre-incubated with different concentrations of NK-EtOAc extract for 30 min, and then stimulated with PMA(50 nM/ml) and A23187($1{\mu}M/ml$) at indicated times. The cell toxicity was determined by MTT assay. The concentrations of prostaglandin E2(PGE2) and cytokines(TNF-${\alpha}$, IL-8) were measured by enzyme-linked immunosorbant assay. Results: NK-EtOAc extract($10{\sim}50{\mu}g/ml$) significantly inhibited the productions of $PGE_2$, TNF-${\alpha}$ and IL-8 in PMA/A23187-stimulated HMC-1 cells without cell toxicity($0{\sim}50{\mu}g/ml$). NK-EtOAc extract also inhibited PMA/A23187-induced phosphorylation of ERK1/2 MAPK and the NF-${\kappa}B$ p65 subunit translocation into the nuclear of HMC-1 cells. Conclusions: This study suggests that NK-EtOAc extract may have an anti-inflammatory property through suppressing the production of inflammatory mediators in activated mast cells and its molecular mechanism underlies the blocking of NF-${\kappa}B$ pathway.

• The Korea Journal of Herbology
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• v.23 no.4
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• pp.91-101
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• 2008

Objectives: In this study, we investigated the inhibitory effects of Ampelopsis Radix methanol(AR-M) extract on allergic inflammation in activated human mast cells and its potential therapeutic or toxic effects. Methods: Ampelopsis Radix(AR) was extracted with 80% methanol. HMC-1 cells, a human mast cell line, were treated with different concentrations of AR-M extract, and then stimulated with PMA plus A23187. The cell toxicity of AR-M extract was determined by MTT assay. The concentrations of $PGE_2$ and cytokines were measured by ELISA. The gene expression of COX-2 and its protein levels were determined by RT-PCR and Western blot. The phosphorylation of ERK MAPK and the NF-${\kappa}B$ activation were determined by Western blot. Results: AR-M extract was significantly inhibited the production of PGE2 and inflammatory cytokines(TNF-${\alpha}$, IL-6 and IL-8) in PMA/A23187-stimulated HMC-1 cells. AR-M extract also attenuated the mRNA expression of COX-2 and its protein induction. Furthermore, AR-M extract attenuated PMA/A23187-induced phophorylation of ERK1/2 MAPK and the NF-${\kappa}B$ p65 subunit translocation into nuclear of HMC-1 cells. AR-M extract significantly decreased PMN A23187-induced release of histamine in a dose-dependent manner. Conclusions: These results indicate that Ampelopsis Radix shows the property of anti-allergic inflammation In vitro through suppressing the production of inflammatory mediators released from mast cells, suggesting have a potential for the treatment of allergic diseases.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.289-296
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• 2014

Human adipose-tissue-derived stromal cells (hADSCs) are abundant in adipose tissue and can differentiate into multi-lineage cell types, including adipocytes, osteoblasts, and chondrocytes. In order to define the optimal harvest site of adipose tissue harvest site, we solated hADSCs from different subcutaneous sites (upper abdomen, lower abdomen, and thigh) and compared their proliferation and potential to differentiate into adipocytes and osteoblasts. In addition, this study examined the effect of phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, on proliferation and differentiation of hADSCs to adipocytes or osteoblasts. hADSCs isolated from different subcutaneous depots have a similar growth rate. Fluorescence-activated cell sorting (FACS) analysis showed that the expression levels of CD73 and CD90 were similar between hADSCs from abdomen and thigh regions. However, the expression of CD105 was lower in hADSCs from the thigh than in those from the abdomen. Although the adipogenic differentiation potential of hADSCs from both tissue regions was similar, the osteogenic differentiation potential of hADSCs from the thigh was greater than that of hADSCs from the abdomen. Phorbol 12-myristate 13-acetate (PMA) treatment increased osteogenic differentiation and suppressed adipogenic differentiation of all hADSCs without affecting their growth rate and the treatment of Go6983, a general inhibitor of protein kinase C (PKC) blocked the PMA effect. These findings indicate that the thigh region might be a suitable source of hADSCs for bone regeneration and that the PKC signaling pathway may be involved in the adipogenic and osteogenic differentiation of hADSCs.

• Biomolecules & Therapeutics
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• v.30 no.6
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• pp.540-545
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• 2022

Betulin is a triterpenoid natural product contained in several medicinal plants including Betulae Cortex. These medicinal plants have been used for controlling diverse inflammatory diseases in folk medicine and betulin showed anti-inflammatory, antioxidative, and anticancer activities. In this study, we tried to examine whether betulin exerts a regulative effect on the gene expression of MUC5AC mucin under the status simulating a pulmonary inflammation, in human airway epithelial cells. Confluent NCI-H292 cells were pretreated with betulin for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h or the indicated periods. The MUC5AC mucin mRNA expression and mucin glycoprotein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. To elucidate the action mechanism of betulin, effect of betulin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated by western blot analysis. The results were as follows: 1) Betulin significantly suppressed the production of MUC5AC mucin glycoprotein and down-regulated MUC5AC mRNA expression induced by PMA in NCI-H292 cells. 2) Betulin inhibited NF-κB activation stimulated by PMA. Suppression of inhibitory kappa B kinase (IKK) by betulin led to the inhibition of the phosphorylation and degradation of inhibitory kappa B alpha (IκBα), and the nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of MUC5AC glycoprotein production in NCI-H292 cells. These results suggest betulin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

• Biomolecules & Therapeutics
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• v.12 no.2
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• pp.79-84
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• 2004

02PS15 extracts (BuOH, $H_2O$, and crude extracts) significantly inhibited IL-4 and IL-6 secretion from the phytohemagglutinin (PHA)-plus phorbol 12-myristate 13-acetate (PMA)-induced peripheral blood mononuclear cleas (P<0.05). 02PS15 extracts (BuOH and crude extracts) also significantly inhibited the histamine release from rat peritoneal mast cells (P<0.05). Significant reduced levels (P<0.05) of PMA- and A23187-induced IL-8 were observed in the human mast cell line, HMC-1, with O2PS15 extracts (BuOH and crude extracts). 02PS15 extracts (BuOH and crude extract) downregulated the expression of IL-6 and IL-8 in the activated HMC-1. These results suggest that O2PS15 has the inhibitory effect of atopic allergic reaction anil this might be useful for clinical application to treat several allergic diseases such as atopic dermatitis.

• Journal of Microbiology and Biotechnology
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• v.30 no.12
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• pp.1835-1842
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• 2020

Ergosterol, an essential constituent of membrane lipids of yeast, is distributed in both the cell membrane and intracellular endomembrane components such as vacuoles. Honokiol, a major polyphenol isolated from Magnolia officinalis, has been shown to inhibit the growth of Candida albicans. Here, we assessed the effect of honokiol on ergosterol biosynthesis and vacuole function in C. albicans. Honokiol could decrease the ergosterol content and upregulate the expression of genes related with the ergosterol biosynthesis pathway. The exogenous supply of ergosterol attenuated the toxicity of honokiol against C. albicans. Honokiol treatment could induce cytosolic acidification by blocking the activity of the plasma membrane Pma1p H+-ATPase. Furthermore, honokiol caused abnormalities in vacuole morphology and function. Concomitant ergosterol feeding to some extent restored the vacuolar morphology and the function of acidification in cells treated by honokiol. Honokiol also disrupted the intracellular calcium homeostasis. Amiodarone attenuated the antifungal effects of honokiol against C. albicans, probably due to the activation of the calcineurin signaling pathway which is involved in honokiol tolerance. In conclusion, this study demonstrated that honokiol could inhibit ergosterol biosynthesis and decrease Pma 1p H+-ATPase activity, which resulted in the abnormal pH in vacuole and cytosol.

• Journal of Life Science
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• v.20 no.3
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• pp.388-395
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• 2010

Cyclooxygenase (COX)-2 is generally known as an inducible enzyme, and it produces arachidonic acid to prostaglandin $E_2$ ($PGE_2$), which has been demonstrated to play a critical role in inflammation. In the present study, we investigated the effects of the extracts of fermented beans including soybean (FS), black agabean (FBA) and yellow agabean (FYA), on the expression of COXs and production of $PGE_2$ in U937 human promonocytic cells. Treatment of phorbol 12-myristate 13-acetate (PMA) significantly induced pro-inflammatory mediators such as COX-2 expression and $PGE_2$ production, whereas the levels of COX-1 remained unchanged. However, pre-treatment with FS, FBA and FYA significantly decreased PMA-induced COX-2 protein as well as mRNA, which is associated with inhibition of $PGE_2$ production. Moreover, FS, FBA and FYA markedly prevented the increase of nuclear translocation of nuclear factor kappa B (NF-${\kappa}B$) p65 by PMA. Our data indicate that the extracts of fermented beans exhibits anti-inflammatory properties by suppressing the transcription of pro-inflammatory cytokine genes through the NF-${\kappa}B$ signaling pathway.