• IMMUNE NETWORK
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• 제6권3호
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• pp.117-122
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• 2006

Background: Caveolin, a family of integral membrane proteins are a principal component of caveolae membranes. In this study, we investigated the effect of p38 kinase on differentiation and on inflammatory responses in sodium nitroprusside (SNP)-treated chondrocytes. Methods: Rabbit articular chondrocytes were prepared from cartilage slices of 2-week-old New Zealand white rabbits by enzymatic digestion. SNP was used as a nitric oxide (NO) donor. In this experiments measuring SNP dose response, primary chondrocytes were treated with various concentrations of SNP for 24h. The time course of the SNP response was determined by incubating cells with 1mM SNP for the indicated time period $(0{\sim}24h)$. The cyclooxygenase-2 (COX-2) and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin $E_2\;(PGE_2)$ assay was used to measure the COX-2 activity. The tyrosine phosphorylation of caveolin-1 was determined by immunoblot analysis and immunostaining. Results: SNP treatment stimulated tyrosine phosphorylation of caveolin-1 and activation of p38 kinase. SNP additionally caused dedifferentiation and inflammatory response. We showed previously that SNP treatment stimulated activation of p38 kinase and ERK-1/-2. Inhibition of p38 kinase with SB203580 reduced caveolin-1 tyrosine phosphorylation and COX-2 expression but enhanced dedifferentiation, whereas inhibition of ERK with PD98059 did not affect caveolin-1 tyrosine phosphorylation levels, suggesting that ERK at least is not related to dedifferentiation and COX-2 expression through caveolin-1 tyrosine phosphorylation. Conclusion: Our results indicate that SNP in articular chondrocytes stimulates dedifferentiation and inflammatory response via p38 kinase signaling in association with caveolin-1 phosphorylation.

• Natural Product Sciences
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• 제17권2호
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• pp.90-94
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• 2011

An in vitro bioassay-guide revealed that the methanol (MeOH) extract of the whole plant of Patrinia saniculaefolia (Valerianaceae) showed cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) dual inhibitory activity by assessing their effects on the production of prostaglandin $D_2$ ($PGD_2$) and leukotriene $C_4$ ($LTC_4$) in mouse bone marrow-derived mast cells (BMMCs). Phytochemical study of the MeOH extract of this plant led to the isolation of twelve compounds; ${\beta}$-farnesene (1), squalene (2), nardostachin (3), patridoid I (4), patridoid II (5), patridoid II-A (6), oleanolic acid (7), oleanonic acid (8), 23-hydroxyursolic acid (9), oleanolic acid 3-O-${\alpha}$-L-arabinopyranoside (10), oleanolic acid 3-O-${\beta}$-D-glucopyranoside (11), oleanolic acid 3-O-[${\beta}$-D-xylopyranosyl-(1${\rightarrow}$3)-${\beta}$-D-(6-O-butyl)glucuronopyranoside] (12). Among the compounds, 4 and 5 strongly inhibited both the COX-2-dependent $PGD_2$ generation with $IC_{50}$ values of 8.7 and 13.6 ${\mu}M$, respectively, and the generation of $LTC_4$ in the 5-LOX dependent phase with $IC_{50}$ values of 41.7 and 46.9 ${\mu}M$, respectively, which suggest that the anti-inflammatory activity of P. saniculaefolia might occur in part via the inhibition of both $PGD_2$ and $LTC_4$ generation by 4 and 5.

• Archives of Pharmacal Research
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• 제26권5호
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• pp.383-388
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• 2003

Bee venom is used as a traditional medicine for treatment of arthritis. The anti-inflammatory activity of the n-hexane, ethyl acetate, and aqueous partitions from bee venom (Apis mellifera) was studied using cyclooxygenase (COX) activity and pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production, in vitro. COX-2 is involved in the production of prostaglandins that mediate pain and support the inflammatory process. The aqueous partition of bee venom showed strong dose-dependent inhibitory effects on COX-2 activity ($IC_{50} = 13.1 \mu$ g/mL), but did not inhibit COX-1 activity. The aqueous partition was subfractionated into three parts by molecular weight differences, namely, B-F1 (above 20 KDa), B-F2 (between 10 KDa and 20 KDa) and BF-3 (below 10 KDa). B-F2 and B-F3 strongly inhibited COX-2 activity and COX-2 mRNA expression in a dose-dependent manner, without revealing cytotoxic effects. TNF-$\alpha and IL-1\beta$ are potent pro-inflammatory cytokines and are early indicators of the inflammatory process. We also investigated the effects of three subfractions on TNF-$\alpha and IL-1\beta$ production using ELISA method. All three subfractions, B-F1, B-F2 and B-F3, inhibited TNF-$\alpha and IL-1\beta$production. These results suggest the pharmacological activities of bee venom on anti-inflammatory process include the inhibition of COX-2 expression and the blocking of pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production.

• Journal of Ginseng Research
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• 제43권2호
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• pp.319-325
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• 2019

Background: Ginsenoside Rf is a ginseng saponin found only in Panax ginseng that affects lipid metabolism. It also has neuroprotective and antiinflammatory properties. We previously showed that Korean Red Ginseng (KRG) inhibited the expression of cyclooxygenase-2 (COX-2) by hypoxia via peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$). The aim of the current study was to evaluate the possibility of ginsenoside Rf as an active ingredient of KRG in the inhibition of hypoxia-induced COX-2 via $PPAR{\gamma}$. Methods: The effects of ginsenoside Rf on the upregulation of COX-2 by hypoxia and its antimigration effects were evaluated in A549 cells. Docking of ginsenoside Rf was performed with the $PPAR{\gamma}$ structure using Surflex-Dock in Sybyl-X 2.1.1. Results: $PPAR{\gamma}$ protein levels and peroxisome proliferator response element promoter activities were promoted by ginsenoside Rf. Inhibition of COX-2 expression by ginsenoside Rf was blocked by the $PPAR{\gamma}-specific$ inhibitor, T0070907. The $PPAR{\gamma}$ inhibitor also blocked the ability of ginsenoside Rf to suppress cell migration under hypoxia. The docking simulation results indicate that ginsenoside Rf binds to the active site of $PPAR{\gamma}$. Conclusions: Our results demonstrate that ginsenoside Rf inhibits hypoxia induced-COX-2 expression and cellular migration, which are dependent on $PPAR{\gamma}$ activation. These results suggest that ginsenoside Rf has an antiinflammatory effect under hypoxic conditions. Moreover, docking analysis of ginsenoside Rf into the active site of $PPAR{\gamma}$ suggests that the compound binds to $PPAR{\gamma}$ in a position similar to that of known agonists.

• 대한본초학회지
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• 제32권2호
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• pp.17-24
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• 2017

Objective : This study was designed to evaluate candidate materials as anti-inflammation agent from extracts of various Korean Compositae herbs in Hwaak mountain. Among Korea medicinal herbs, Ainsliaea acerifolia (AA) belongs to the Compositae family, has been used for the treatment of rheumatic arthritis. However, AA has not been previously reported to have an anti-inflammatory effect. Therefore, we investigated the anti-inflammatory effects of AA and its underlying molecular mechanisms in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Methods : Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in RAW 264.7 macrophages. Nitric oxide (NO) was measured with Griess reagent and pro-inflammatory cytokines were detected by enzyme immunoassay (EIA) kits in LPS-stimulated RAW 264.7 macrophages. Protein expressions of inducible nitric oxide synthase, and cyclooxygenase-2 (COX-2) and p65 subunit of nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) were determined by Western blot analysis. Results : Among 8 extracts of Korean Compositae herbs tested, AA showed the inhibition of NO production without cytotoxicity. Consistent with the observation, AA reduced the expression levels of iNOS and COX-2 proteins in LPS-simulated RAW 264.7 macrophages in dose-dependent manner. In addition, AA inhibited the productions of $TNF-{\alpha}$ and IL-6 in LPS-simulated RAW 264.7 macrophages. However, AA did not inhibit activation of p65 $NF-{\kappa}B$ in LPS-simulated RAW 264.7 macrophages. Conclusion : These results suggest that down-regulation of iNOS, COX-2 protein expression and $TNF-{\alpha}$ and IL-6 production by AA are responsible for its anti-inflammatory effects.

• 한국식품과학회지
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• 제39권3호
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• pp.348-352
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• 2007

한국인이 상용하는 식물성 식품 30종의 에탄올 추출물을 이용하여 $sPLA_{2}$, COX-1, COX-2, 12-LOX 의 활성 억제 효과를 측정하여 식물 추출물이 염증 효소계에 미치는 영향을 포괄적으로 평가하였다. 그 결과 모든 식물 추출물은 적어도 한 개 이상의 염증 관련 효소의 활성을 저해하였다. 몇 가지 두류와 숙주나물, 일부 잎채소는 염증반응의 상위 단계 효소인 $sPLA_{2}$의 활성을 저해하였으며, 12-LOX 활성은 발아나물인 콩나물과 숙주나물에 의해서만 특이적으로 저해되었다. 대부분의 식품들은 COX-1과 COX-2 활성을 동시에 저해하였고, 미나리와 비름만이 COX-1 활성 저해 없이 COX-2만을 선택적으로 저해하였다. 모든 두류 유래식품과 식물 뿌리류는 COX-2에만 선택적이지는 않았으나, COX-1보다 더 낮은 농도에서 COX-2 활성을 억제하였다. 여러 식품들 중 염증반응의 상위단계 효소인 $sPLA_{2}$ 활성을 억제하는 일부 두류와 잎채소류 및 숙주나물은 염증 초기에 작용하여 염증 반응의 발전을 차단하는데 도움을 줄 수 있을 것으로 보인다. 특히, 녹두는 비교적 낮은 $IC_{50}$ 값을 보이며 $sPLA_{2}$와 COX-2를 효과적으로 저해하는 것으로 나타나 염증반응의 여러 단계에서 항염 효과를 나타낼 수 있는 유용한 식품으로 판단된다. 또한 각각의 염증 관련 효소에 대한 억제 능력이 크지는 않았지만, 염증 반응의 초기 및 후기 단계의 모든 효소를 저해 하였던 숙주나물과 12-LOX 및 COX-2를 동시에 저해한 콩나물도 여러 염증 효소를 복합적으로 억제시킴으로써 항염능을 나타낼 수 있다는 점에 주목 할 필요가 있다. 본 연구 결과 식물성 식품에 의한 염증 완화 및 예방 효과는 각기 다른 염증 효소의 활성을 다양한 정도로 저해함으로써 발현됨을 알 수 있다. 밝혀졌으며 제2형 당뇨모델인 $KK-A^{y}$ 마우스를 이용한 동물실험에서도 뚜렷한 혈당강하효과를 나타내어 인슐린 민감성 제재로 개발 할 수 있는 가능성을 보여 주었다.균은 $0.9{\sim}2.6%$이었으며, 8종류 약제에 저항성인 균도 1.7%있었다. 이상의 결과로 국내에서 분리된 M. pneumoniae 균주는 적게는 1-4 종류의 항생제에, 많게는 5-8 종류의 항생제에 저항성인 균주가 있으므로 마이코플라스마폐렴 환자를 치료할때는 macrolide계나 quinolone계의 항생제 선택에 신중을 기하여야 하며, 가급적이면 항생제 감수성 검사를 실시하여 적절한 항생제를 선택함으로써 저항성균의 출현율을 줄일 수 있고 효율적인 치료도 할 수 있도록 하여야 할 것으로 생각된다.이어트에도 도움이 되리라 생각한다. 56.3%, 엽산 81.3% 등으로 높게 나타나 근로자들의 영양 문제가 심각함을 알 수 있다.혁신지방분권위원회에서 제시한 자치경찰제도(안)을 중심으로 자치경찰제도 운용의 목적 충족과 실질적인 효과의 측면에서 분석하고 바람직한 자치경찰제도의 운용에 대해 살펴본다.rc}C$ 이내에서 높을수록, 염분은 20-35 psu 이내에서 높을수록 잠입률이 높은 경향을 나타내었다. 교수학습모형에 관련된 지식을 묻는 내용으로 주로 출제되었다. 이에 구체적인 개선방안으로 특정 교수학습모형의 이론적 토대가 되고 전체적인 교수설계를 하기 위한 기본 바탕이 될 수 있는 교수학습이론에 관한 내용, 또한 현재가정과교육에 있어서 유용한 교수학습법이라고 입증되고 있는 실천적 추론 가정과 수업에 관한 내용으로의 확대를 제안하였다. 가정과교육평가 문항의 출제는 대다수의 문항이 수행평가에 관한 문항내용으로 출제되었다. 이에 구체적인 개선방안으로 문항의 변별도 여부의 판단, 평가문항의 내용 타당도 분석, 평가결과를 해석하는 능력, 평가자의 철학적 관점과 같은 내용으로의 확대를 제안하였다.

• Archives of Pharmacal Research
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• 제26권7호
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• pp.545-553
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• 2003

The expression of cyclooxygenase-2 (COX-2) is a characteristic response to inflammation, which can be inhibited with sodium salicylate. IL-1$\beta$ and TNF-$\alpha$ can induce extracellular signal-regulated kinase (ERK), IKK, IkB degradation and NF-$\kappa$B activation. Salicylate inhibited the IL-1$\beta$ and TNF-$\alpha$-induced COX-2 expressions, regulated the activation of ERK, IKK and IkB degradation, and the subsequent activation of NF-$\kappa$B, in neonatal rat ventricular cardiomyocytes. The inhibition of the ERK pathway, with a selective inhibitor, PD098059, blocked the expressions of IL-1$\beta$ and TNF-$\alpha$-induced COX-2 and $PGE_2$ release. The antioxidant, N-acetyl-cysteine, also reduced the glutathione or catalase- attenuated COX-2 expressions in IL-1$\beta$ and TNF-$\alpha$-treated cells. This antioxidant also inhibited the activation of ERK and NF-$\kappa$B in neonatal rat cardiomyocytes. In addition, IL-1$\beta$ and TNF-$\alpha$-stimulated the release of reactive oxygen species (ROS) in the cardiomyocytes. However, salicylate had no inhibitory effect on the release of ROS in the DCFDA assay. The results showed that salicylate inhibited the activation of ERK and IKK, I$\kappa$B degradation and NF-$\kappa$B activation, independently of the release of ROS, which suggested that salicylate exerts its anti-inflammatory action through the inhibition of ERK, IKK, IkB and NF-$\kappa$B, and the resultant COX-2 expression pathway in neonatal rat ventricular cardiomyocytes.

• The Korean Journal of Physiology and Pharmacology
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• 제7권4호
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• pp.239-246
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• 2003

The expression of cyclooxygenase-2 (COX-2) is a characteristic response to inflammation and can be inhibited with sodium salicylate. $TNF-{\alpha}$ plus $IFN-{\gamma}$ can induce extracellular signal-regulated kinase (ERK), IKK, $I{\kappa}B$ degradation and NF-${\kappa}B$ activation. The inhibition of the ERK pathway with selective inhibitor, PD098059, blocked cytokine-induced COX-2 expression and $PGE_2$ release. Salicylate treatment inhibited COX-2 expression induced by $TNF-{\alpha}$/$IFN-{\gamma}$ and regulated the activation of ERK, IKK and $I{\kappa}B$ degradation and subsequent NF-${\kappa}B$ activation in MC3T3E1 osteoblasts. Furthermore, antioxidants such as catalase, N-acetyl-cysteine or reduced glutathione attenuated COX-2 expression in combined cytokines-treated cells, and also inhibited the activation of ERK, IKK and NF-${\kappa}B$ in MC3T3E1 osteoblasts. In addition, $TNF-{\alpha}$/$IFN-{\gamma}$ stimulated ROS release in the osteoblasts. However, salicylate had no obvious effect on ROS release in DCFDA assay. The results showed that salicylate inhibited the activation of ERK and IKK, $I{\kappa}B$ degradation and NF-${\kappa}B$ activation independent of ROS release and suggested that salicylate exerts its anti-inflammatory action in part through inhibition of ERK, IKK, $I{\kappa}B$, $NF-{\kappa}B$ and resultant COX-2 expression pathway.

• 대한한방부인과학회지
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• 제20권4호
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• pp.56-73
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• 2007

Purpose: 이 실험은 골다공증의 치료약물로 자하거의 골질재흡수 억제효과를 검토하기 위하여 설계되었다. Methods: 자하거의 골질재흡수 효과를 확인하기 위하여 생쥐의 두개골 골모세포를 이용하여 Cyclooxygenase-1(COX-1), COX-2, $TGF-{\beta}$, $L-1{\beta}$, $TNF-{\alpha}$, IL-6, prostaglandin E2등의 활성화 정도를 측정하였으며, 골조직의 미세구조적 변화를 확인하였다. Results: 자하거는 $IL-1{\beta}$, $TNF-{\alpha}$, IL-6 또는 그 세가지의 조합에 의하여 유발된 PGE2의 생성 뿐만 아니라 COX-2 mRNA 수치도 감소시켰으나 COX-1 mRNA 수치에는 영향을 주지 않았다. 이로써 자하거는 시험관내에서 그리고 생체내에서 펩티드의 인산화를 억제함으로써 골의 재흡수를 저해하였다. 그리고 자하거는 생쥐에서 $IL-1{\beta}$에 의해 유발된 고칼슘혈증을 감소시켰고, 골의 재흡수를 저해하는 경로를 통하여 골에 대한 보호효과를 보여줌으로써 조기에 난소 절제한 쥐에서 골질감소와 미세구조적 변화를 부분적으로 방지하였다. 이러한 결과는 PGE2 생성에 대한 $IL-1{\beta}$, $TNF-{\alpha}$, IL-6사이의 상승효과는 COX-2의 유전자 발현이 증가한 결과이며 이러한 tyrosine kinase가 생쥐의 두개골 골모세포에서 COX-2의 신호전달에 관계한다는 것을 보여준다. Conclusion: 자하거가 생쥐의 두개골 골모세포에서 여러 신호전달물질의 활성화를 통하여 골질재흡수를 저해하는 특성을 확인함으로써 앞으로 골다공증의 예방과 치료에 대한 추가적인 임상연구가 필요할 것으로 사료된다.

• Animal cells and systems
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• 제16권5호
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• pp.366-375
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• 2012

Gallotannin (GT) is derived from plant poly phenol and is associated with biological actions in a wide range of cells. In this study, we evaluated the effect of GTon apoptosis and cyclooxygenase-2 (COX-2) expression and attempted to shed light on the mechanism of action in A549 human lung carcinoma cells. We found that GT dramatically induced apoptosis as demonstrated by expression of p53 and active caspase-3 via western blot analysis and fragmented DNA as detected by DNA fragmentation and DAPI staining. We also observed that GT significantly causes COX-2 expression in a dose-dependent manner determined by western blot analysis. Phosphorylation of Akt and p38 was considerably increased by GT in A549 human lung carcinoma cells. Inhibition of Akt and p38kinase with LY294002 or SB203580 suppressed GT-induced apoptosis and COX-2 expression. Furthermore, we have shown that prevention of COX-2 with NS398 or indomethacin does not any effects on apoptosis induced by GT. Taken together, our present results suggest that GT regulates apoptosis and COX-2 expression through Akt and p38kinase pathway in A549, human lung carcinoma cells.