• Natural Product Sciences
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• 제22권3호
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• pp.147-153
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• 2016

Inflammation plays an important role in host defense against external stimuli such as infection by pathogen, endotoxin or chemical exposure by the production of the inflammatory mediators that produced by macrophage. Anti-inflammatory factor is important to treat the dangers of chronic inflammation associated with chronic disease. This research aims to analyze the anti-inflammatory effects of Garcinia mangostana L. peel extract (GMPE), ${\alpha}$-mangostin, and ${\gamma}$-mangostin in LPS-induced murine macrophage cell line (RAW 264.7) by inhibiting the production of inflammatory mediators. The cytotoxic assay of G. mangostana L. extract, ${\alpha}$-mangostin, and ${\gamma}$-mangostin were performed by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) to determine the safe and non-toxic concentration in RAW 264.7 for the further assay. The concentration of inflammatory mediators (COX-2, IL-6, and IL-$1{\beta}$) were measured by the ELISA-based assay and NO by the nitrate/nitrite colorimetric assay in treated LPS-induced RAW 264.7 cells. The inhibitory activity was determined by the reducing concentration of inflammatory mediators in treated LPS-induced RAW 264.7 over the untreated cells. This research revealed that GMPE, ${\alpha}$-mangostin, and ${\gamma}$-mangostin possess the anti-inflammatory effect by reducing COX-2, IL-6, IL-$1{\beta}$, and NO production in LPS-induces RAW 264.7 cells.

• 생명과학회지
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• 제21권10호
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• pp.1377-1384
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• 2011

Astaxanthin (ATX)은 다양한 생명체에서 생성되는 카로티노이드 색소이다. 본 연구에서는 ATX가 RAW264.7 cell에서 LPS에 의한 inducible nitric oxide synthase (iNOS), nitric oxide (NO), 염증성 사이토카인, nuclear factor-kappa B (NF-${\kappa}B$)와 reactive oxygen species (ROS)의 생성을 억제 시키는 지 또한, superoxide radical 소거능이 있는 지를 조사하였다. iNOS와 NF-${\kappa}B$는 immunoblot analysis로, interleukin (IL)-6와 tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$)는 ELISA 법으로 분석하였다. NO 양은 nitrite의 양을 측정하였고, ROS는 2',7'-dichlorodihydrofluorescin diacetate (DCFH-DA) 법으로 superoxide radical 소거능은 superoxide radical scavenging activity assay로 검증하였다. 100 ${\mu}M$의 ATX 농도에서 LPS로 유도된 NO, IL-6 및 TNF-${\alpha}$ 같은 염증성 사이토카인의 생성 뿐만 아니라 iNOS 및 NF-${\kappa}B$의 발현도 억제되었다. 특히, IL-6 및 TNF-${\alpha}$ 생성에 있어 ATX의 최대 억제율은 각각 65.2% 및 21.2% 이었으며 LPS로 유도된 NF-${\kappa}B$의 전사활성을 억제하였다. 이러한 현상은 세포질에서 핵으로 NF-${\kappa}B$의 전위를 억제하는 것과 관련이 있다. 또한, 25-100 ${\mu}M$의 ATX 농도에서 세포 내 ROS 생성을 억제하였으며, 5 mg/ml 농도의 ATX는 동일농도의 ${\alpha}$-tocopherol에 비해 superoxide radical 소거능이 1.33배 높았다. 이러한 결과들은 ATX가 대식세포에서 ROS 생성 및 NF-${\kappa}B$ 활성을 저해하므로 iNOS의 발현, NO 및 염증성 사이토카인의 생성을 억제하며, 또한 우수한 superoxide radical 소거능을 보유한다는 것을 나타내었다. 결론적으로, ATX가 항염증제 및 항산화제로서 유용하게 사용될 수 있을 것으로 사료된다.

• Molecular & Cellular Toxicology
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• 제1권3호
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• pp.179-188
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• 2005

To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.

• 한국식품저장유통학회지
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• 제22권4호
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• pp.613-621
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• 2015

본 연구에서는 마늘의 부산물로 발생하는 마늘대의 항산화 및 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 Raw 264.7 세포에 대한 열수추출물(ASSW)과 70% 에탄올 추출물(ASSE)의 효과를 살펴보았다. ASSW의 폴리페놀 함량은 $37.08{\pm}1.51mg(TAE)/g$, ASSE의 폴리페놀 함량은 $44.7{\pm}1.32mg(TAE)/g$ 이 함유되어있음을 확인하였다. DPPH 실험과 $ABTS^+$ 실험에서 ASSW, ASSE 모두 농도 의존적으로 증가하였으며, DPPH의 경우 $1,000{\mu}g/mL$에서 대조군인 Vit.C의 $50{\mu}g/mL$의 항산화능이 있다는 것이 확인되었고, $ABTS^+$의 경우 $500{\mu}g/mL$ 이상부터 대조군인 Vit.C와 비슷한 효과를 나타냄으로서 ASSW, ASSE 모두 항산화능이 있다는 것을 확인하였다. MTT측정으로 인해 세포 독성을 가지지 않았던 농도대(5, 10, 25, 50, $100{\mu}g/mL$)에서 염증 억제 효과를 살펴보기 위해 NO를 측정한 결과 ASSW, ASSE 모두 $25{\mu}g/mL$에서부터 유의적으로 분비량이 감소함을 확인하였으며 특히 $100{\mu}g/mL$의 농도에서 약 18%, 23%의 억제 효과를 보였다. 또한 대식세포의 염증성 cytokine인 IL-6, TNF-${\alpha}$, IL-$1{\beta}$ 및 $PGE_2$의 분비량을 첨가 농도 의존적으로 억제함을 확인하였다. 특히 $PGE_2$에 대해 ASSW, ASSE $100{\mu}g/mL$의 농도에서 약 55%, 60%의 감소효과를 보였다. ASSW의 iNOS, COX-2의 발현 저해효과는 나타내지 못하였지만, ASSE는 $100{\mu}g/mL$의 농도에서 iNOS의 발현량이 현저하게 억제됨을 확인하였고, COX-2의 경우 농도 의존적으로 저하되어 특히 $50{\mu}g/mL$, $100{\mu}g/mL$의 구간에서 단백질 발현 저해효과가 있음을 확인하였다. 이를 통해 ASSW, ASSE 모두 항산화 효과와 항염증 효과가 있음을 확인하였으며, ASSW 보다 ASSE에서 활성산소종(reactive oxygen species, ROS) 및 ROS에 의해 유발되는 염증을 억제시켜주는 소재가 될수있을 것이라 예상된다.

• 대한한의학회지
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• 제39권2호
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• pp.92-105
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• 2018

Objectives: This aim of this study is to compare the reflux esophagitis improvement effects of Sepiae Os, Arcae Concha, Ostreae Concha, and Proton Pump Inhibitor(esomeprazole) through rat experiments. Methods: NO production inhibitory effect was measured by NO production amount and iNOS mRNA expression level in cell lines. iNOS, $TNF-{\alpha}$ and $p-I{\kappa}B$, and serotonin were compared using immunohistochemistry at the rat reflux esophagitis. Reflux esophagitis connection external form, lower esophageal sphincter, and gap were observed and an esophageal inflammatory indicator, IL-6 activity was also evaluated by immunohistochemistry. Results: NO production and iNOS mRNA expression was showed concentration dependent decrease in cell lines treated with Sepiae OS, Arcae Concha, and Ostreae Concha at the experiments of cell lines. In the suppression of iNOS and $p-I{\kappa}B$ at the rat reflux esophagitis, Sepiae Os treat group(SOT) and Ostreae Concha treat group(OCT) were more effective. In the increase of serotonin at the rat reflux esophagitis, ACT, MT and OCT were more effective. Damage of lower esophageal sphincter, and gap between esophageal keratin and mucosa were observed less at the SOT, ACT, OCT. In the suppression of IL-6 at the rat reflux esophagitis, SOT and OCT were more effective than GE and, SOT was more effective than MT significantly. Conclusions: The anti-inflammatory effect was the best in the SOT and lower esophageal sphincter muscle contraction was the best in the ACT at the rat reflux esophagitis. Sepiae OS was more effective than esomeprazole in the suppression of iNOS, $TNF-{\alpha}$, and IL-6.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.385-393
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• 2008

As an attempt to search for bioactive natural products exerting anti-inflammatory activity, we evaluated the effects of the methanol extract of Polytrichum commune Hedw (PCM) (Polytrichaceae) on lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines release in murine macrophage cell line RAW 264.7. PCM potently inhibits the production of NO, $PGE_2$, tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Consistent with these results, PCM also concentration-dependently inhibited LPS-induced inducible NO synthase (iNOS) and cyclooxygase (COX)-2 at the protein levels, and iNOS, COX-2, TNF-$\alpha$ and IL-6 at the mRNA levels without an appreciable cytotoxic effect on RAW 264.7 macrophag cells. Furthermore, PCM inhibited LPS-induced nuclear factor-kappa B (NF-$\kappa$B) activation as determined by NF-$\kappa$B reporter gene assay, and this inhibition was associated with a decrease in the nuclear translocation of p65 and p50 NF-$\kappa$B. Taken together, these results suggest that PCM may play an anti-inflammatory role in LPS-stimulated RAW 264.7 macrophages through the inhibitory regulation of iNOS, COX-2, TNF-$\alpha$ and IL-6 via NF-$\kappa$B inactivation.

• 생명과학회지
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• 제16권6호
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• pp.989-993
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• 2006

우리나라 전국 산지에서 쉽게 구할 수 있는 노루발풀을 건조시켜 80% ethanol 추출물과 여러 가지 용매 분획물의 미생물 증식 억제 효과에 대해서 검색하였다. 노루발풀의 80% ethanol 추출물은 그람 양성균 B. subtilis에서 농도 의존적으로 항균활성을 나타내는 반면 그람 음성균 E. coli에서는 활성이 나타나지 않았다. 노루발풀의 80% ethanol 추출물을 용매 분획하여 얻은 분획물 중 hexane층에서 항균활성이 나타나는 것을 확인하였다. 노루발풀의 항균활성 물질을 검색하기 위한 목적으로 hexane 분획물을 silica gel column chromatography, prep. TLC 및 HPLC를 실시하였으며 그 결과, peak I의 단일 물질이 노루발풀의 B. subtilis에 대한 항균활성을 나타내는 물질임을 확인할 수 있었다.

• Journal of Photoscience
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• 제9권2호
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• pp.201-204
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• 2002

Epidermal cells produce a panel of antioxidants as well as cytokines after UVB irradiation, which counteract reactive oxygen species, however, how these antioxidants might regulate melanogenesis is unclear. An important constituent of the cellular antioxidant buffering system which controls the redox state of proteins is thioredoxin (TRX), a 13-kD protein that catalyzes thiol-disulfide exchange reactions, regulates activation of transcription factors, and possesses several other biological functions similar to cytokines. TRX suppressed the UVB-induced production and secretion of $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH) and of adrenocorticotropic hormone (ACTH), and also suppressed proopiomelanocortin (POMC) mRNA expression by normal human keratinocyte (KC)s. Further, L-cysteine, N-acetyl-cysteine, $\alpha$-tocopheryl ferulate showed suppressive effect on UVB-induced POMC mRNA expression. However, TRX released from UVB-irradiated KCs stimulated melanogenesis by up-regulating MSH receptor expression and its binding activity in melanocyte (MC)s. UVB-induced KC derived cytokines such as IL1, IL6, and ET1 upregulated MSH-receptor binding ability as well as MCl-R mRNA expression in cultured normal human MCs. MCl-R has a tendency to be upregulated by UVB-induced KC-derived cytokines as well as by direct UVB irradiation. These results suggest that antioxidants such as TRX suppresses UVB induction of POMC, but in the case of MCl-R, this gene can be mainly in the trend of upregulation by UVB-induced KC-derived factors including TRX.

• 대한한의학회지
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• 제24권3호
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• pp.84-95
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• 2003

Objective : This study was carried out to investigate the effects of Kamisopunghwalhyeol-tang (Jiaweishujenghuoxie-tang; Kami-SPHHT) on the immunity responses of the Synoviocytes isolated from the patients on rheumatoid arthritis. Methods : Cells were stimulated by $Interleukin-1{\beta}$ and Tumor Necrosis $Factor-{\alpha}$ in the presence or absence of Kami-SPHHT, and then induced cytokine mRNA levels were determined by RT-PCR and real-time quantitative RT-PCR. Results : Levels of $IL-1{\beta},{\;}IL-6,{\;}TNF-{\alpha}$, COX-2, and NOS II mRNA expressions significantly decreased in Kami-SPHHT treated cells compared to non-treated control cells. Also, DNA-binding activity of $NF-{\kappa}B$ and AP-l decreased in Kami- SPHHT treated hFLSs. Conclusion : These results suggest that Kami-SPHHT may be involved in anti-inflammatory reactions by inducing cytokine gene expression in synoviocytes, and further in vivo examination on its efficacy can provide potential application for the treatment of rheumatoid arthritis.

• The Korean Journal of Physiology and Pharmacology
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• 제23권1호
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• pp.29-35
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• 2019

Decursin is a major biological active component of Angelica gigas Nakai and is known to induce apoptosis of metastatic prostatic cancer cells. Recently, other reports have been commissioned to examine the anticancer activities of this plant. In this study, we evaluated the inhibitory activity and related mechanism of action of decursin against glioblastoma cell line. Decursin demonstrated cytotoxic effects on U87 and C6 glioma cells in a dose-dependent manner but not in primary glial cells. Additionally, decursin increased apoptotic bodies and phosphorylated JNK and p38 in U87 cells. Decursin also down-regulated Bcl-2 as well as cell cycle dependent proteins, CDK-4 and cyclin D1. Furthermore, decursin-induced apoptosis was dependent on the caspase activation in U87 cells. Taken together, our data provide the evidence that decursin induces apoptosis in glioblastoma cells, making it a potential candidate as a chemotherapeutic drug against brain tumor.