• 한국약용작물학회지
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• 제20권5호
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• pp.381-386
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• 2012

Eucommia ulmoides OLIVER (EU) is a traditional Korean herbal used for the treatment of rheumatoid arthritis (RA). In the present study, the molecular pharmacology basis of its anti-inflammatory effect is revealed in this work, EU was studied in lipopolysaccharide (LPS)-activated macrophage cells (RAW 264.7) as an established inflammation model. After activation, nitric oxide (NO) production and iNOS mRNA were measured by using a colorimetric assay (Griess reagent), and reverse transcription polymerase chain reaction (RT-PCR), respectively. The change in the content of $PGE_2$, $TNF{\alpha}$, and IL-6 was concurrently monitored by ELISA. In results, we found that in the concentration range without showing cytotoxicity, EU produced a remarkable anti-inflammatory effect and showed a dose-dependent inhibition of LPSinduced NO production. Compared with indomethacin, EU has more potency and a specific action of NO inhibition, $PGE_2$, IL-6, and TNF-${\alpha}$ inhibition. These results suggest that EU may be a suitable herbal medicine to yield the greatest anti-inflammatory activity for food additives and medicine.

• Asian Pacific Journal of Cancer Prevention
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• 제13권3호
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• pp.991-994
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• 2012

Objective: VEGF-C has recently been identified as a key molecule which is involved in tumor lymphangiogenesis. The aim of this research was to investigate the role of PARP-1 inhibition in the regulation of VEGF-C expression in CT26 cells. Methods: CT26 cells were treated with or without the PARP-1 inhibitor 5-aminoisoquinolinone (5-AIQ). The expression of PARP-1, NF-kB, and VEGF-C proteins in CT26 cells was measured by Western blot analysis and the VEGF-C mRNA level was determined by reverse transcription polymerase chain reaction (RT-PCR). CT26-secreted VEGF-C was detected by enzyme-linked immunosorbent assay (ELISA). Results: The results of Western blot analysis showed that the expression levels of PARP-1, NF-kB, and VEGF-C were reduced in 5-AIQ treated CT26 cells and the levels of VEGF-C mRNA in 5-AIQ treated CT26 were significantly lower than t in 5-AIQ-untreated cells (P<0.05). The concentrations of CT26-secreted VEGF-C were also dramatically decreased (P<0.05). Conclusion: Here, we provide evidence for the first time that PARP-1 inhibition dramatically reduces VEGF-C expression via the nuclear factor NF-kB signaling pathway. We therefore propose that PARP-1 inhibition has an anti-lymphangiogenic effect and may contribute to the prevention of metastatic dissemination via the lymphatic system.

• 대한본초학회지
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• 제30권5호
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• pp.67-74
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• 2015

Objectives : TheHaedokgumhwa-sanwater extract (HDKHS) is used in Korea, Japan and China as a traditional therapeutic agent to cure an infectious disease. But its study is not enough. Therefore, the present study focused on the elucidation of HDKHS to investigate the anti-inflammatory effects and to established the possible mechanisms involved in its action on LPS-stimulated immune response in murine macrophages.Methods : Inflammatory status was induced by LPS and measured by increasement of inflammatory mediators. LPS induced secretions of NO and PGE₂in RAW 264.7 cells were measured using griess reagent and enzyme-linked immunosorbent assay (ELISA) kit respectively. production of IL-6 was examined using ELISA kit and expression of IL-6 mRNA was measured by RT-PCR method. To investigate the effects of HDKHS on inflammatory mediators, such as iNOS, COX-2 and MAPKs, western blot and RT-PCR were performed.Results : HDKHS significantly reduced production of NO and PGE2 which were induced by LPS. Also, activation of IL-6 was reduced both protein and mRNA levels. The expressions of inflammatory mediator include iNOS and COX-2 were decreased by pretreatment with HDKHS. futhermore The result showed HDKHS down-regulate the LPS induced phosphorylation of ERK 1/2, one of the MAPK family, which is considered as a main regulator of transmission from pathogens to nucleus of immune cells.Conclusions : Our results suggest that the anti-inflammatory properties of HDKHS may stem from the inhibition of pro-inflammatory mediators via suppression of initiation of inflammatory response by inhibiting MAPKs signaling pathways.

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.145-158
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• 1993

The oral microbiota such as P. gingivalis, P. intermedia and A. actinomycetemcomitans play a primary role in the initiation and progression of the periodontal disease. The purpose of this study was to evaluate the antimicrobial effects and inhibitory effects of honokiol and magnolol on the bacterial collagenase activity, cytotoxicity and cytokine production of periodontopathic microorganisms. The antimicrobial activities of honokiol and magnolol was evaluted with minimum inhibition concentration. Honokiol was more active than magnolol, but less than chlorhexidine on antimicrobial activity. The inhibitory effects of magnolol and honokiol on the collagenolytic activity and cytotoxicity were evaluated using a Collagenokit CLN-100 and rapid colorimetric assay (MTT method) for cellular growth and survival of gingival fibroblast and periodontalligament cell and $[^3H]-thymidine$ incorporation for the gingival epithelial cell. The inhibitory effects on the collagenolytic activity was the highest in chlorhexidine, and the lowest in magnolol. Magnolol had the lowest cytotoxic effect and chlorhexidine had the highest. The inhibitory effects on cytokine production was evaluated using $interleukin-1{\beta}$ ELISA kit (Cistron Biotech.), IL-6, $TNF-{\alpha}$ ELISA kit (Genzyme) and inhibitory effects were higher than bacterial LPS and there is no difference among the honokiol, magnolol and chlorhexidine. From these results, the antimicrobial and antienzymatic activities of honokiol and magnolol were seemed to inhibit bacterial growth and enzyme activities with lesser cytotoxic activities. Therefore, it was suggested that honokiol and magnolol are very effective antimicrobial agents on periodontal pathogens.

• 한국식품영양과학회지
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• 제33권7호
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• pp.1169-1174
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• 2004

본 연구에서는 감마선 및 전자선종이 ovalbumin(OVA)의 구조 변화에 미치는 영향을 비교 평가하였다. 표준항원 OVA을 3,5,7 및 10 kGy의 흡수선량으로 감마선 및 전자선 조사하였으며 그에 따른 구조적인 변화는 SDS-PAGE, GPC-HPLC 및 단클론 항체를 사용한 Ci-ELISA법으로 측정하였다. Native OVA분자는 감마선 및 전자선 조사에 의해 소편화 및 응집화 되었으며, 조사 선량이 증가할수록 OVA의 분자량이 감소하였다. 또한 OVA의 면역화학적 인 구조는 방사선 조사에 의해 항원-항체간의 결합능이 감소하는 것으로 나타났으나 감마선 및 전자선종에 따른 차이는 없었다. 이상의 결과들은 식품 알레르겐의 제거 및 면역원의 개발에 대한 전자선 조사의 기초자료로 이용될 수 있을 것으로 사료된다.

• 한국융합학회논문지
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• 제8권1호
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• pp.239-243
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• 2017

본 연구는 프랑킨센스를 이용하여 전염증성 사이토카인의 분비 억제 효과를 입증하고자한다. 이에 LPS를 처리한 마우스의 대식세포를 이용하여 $TNF-{\alpha}$, $IL-1{\beta}$와 같은 전염증성 사이토카인의 분비 변화를 살펴보기 위해 다자인하였다. 세포에 대한 독성 실험은 MTS인 CellTiter 96 AQueous One solution cell proliferation 시약을 이용하여 실시하였으며 전염증성 사이토카인의 분비는 ELISA kit을 사용하여 측정하였다. 그 결과 프랑킨센스는 10ug/ml-1000ug/ml에서 세포독성이 없었으며 전염증성 사이토카인인 $TNF-{\alpha}$, $IL-1{\beta}$생성을 유의성있게 억제시켰다. 전염증성 사이토카인의 분비 억제효과는 프랑킨센스의 항 염성을 입증하기 위한 기초적인 생리 활성 자료와 기능성 소재로 다양한 융합적인 활용이 가능하다고 사료된다.

• 한방재활의학과학회지
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• 제23권3호
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• pp.27-35
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• 2013

Objectives The aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. Methods Cells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines ($PGE_2$, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results MGH inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. Conclusions These results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

• IMMUNE NETWORK
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• 제7권2호
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• pp.95-100
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• 2007

Background: A red seaweed, Callophyllis japonica has been traditionally eaten in the oriental area. In a recent study, it has been demonstrated that the ethanol extract of C. japonica have antioxidant activity. However, there are few studies about the effects of C. japonica on the function of immune cells. We investigated the immunomodulatory effects of C. japonica on the function of dendritic cells, the potent antigen-presenting cells. Methods: Bone marrow-derived dendritic cells (DCs) were used and the viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and trypan blue exclusion test. Cytokine and nitric oxide (NO) levels were determined by using ELISA and Griess reagent, respectively. The expression levels of DC surface markers were measured by flow cytometric analysis. Results: C. japonica ethanol extract did not significantly affect the DCs viability and the IL-12 production from DCs, irrespective of the presence of lipopolysaccharide (LPS). In addition, it did not significantly change the expression of DC surface markers. However, C. japonica ethanol extract significantly inhibited the LPS-induced NO production and also increased the proliferation of allogeneic lymphocytes activated by DCs. Conclusion: Our data suggests that C. japonica ethanol extract enhances the proliferation of allogeneic lymphocytes activated by DCs which is associated with inhibition of NO production from DCs induced by LPS.

• 대한본초학회지
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• 제21권4호
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• pp.69-76
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• 2006

Objectives : The fresh young leaves and dried fruits of Zanthoxylum piperitum (Korean name: Chopi) are used as diuretics, stomachies, anthelmintic and for the treatments of disorders of the digestive organ in Asia. We investigated inhibitory effects of Zanthoxylum piperitum extract on lipopolysaccharide(LPS)-induced production of nitric oxide(NO) and pro-inflammatory cytokines including $TNF-{\alpha}$ and $IL-1{\beta}$ from RAW264.7 mouse macrophage cells. Methods : After methanol extract of Zanthoxylum Fructus (Zanthoxylum extract) was pretreated in RAW264.7 cells, the cells were stimulated with LPS. Cell toxicity of Zanthoxylum extract was assayed bv MTT assay. The production of NO from the cells was measured in culture medium by Griess reaction. The production of $TNF-{\alpha}$ and $IL-1 \;{\beta}$ from the cells was measured in culture medium by ELISA. Results : Zanthoxylum Fructus extract greatly inhibited the production of inflammatory mediators such as NO, $TNF-{\alpha}$ and $IL-1{\beta}$ from LPS-stimulated RAW264.7 cells. Conclusion : This result suggests that Zanthoxylum extract may have an anti-inflammatory effect through the inhibition of inflammatory mediators.

• BMB Reports
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• 제37권6호
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• pp.726-734
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• 2004

White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.