• 대한한의진단학회지
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• 제17권3호
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• pp.263-273
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• 2013

Objectives This study was designed to investigate the anti-inflammatory effect of extracts from Ligustrum obtusifolium S. fruits(LOF) in RAW 264.7 Macrophages stimulated with lipopolysaccharide(LPS). Methods We examined productions of nitric oxide(NO), reactive oxygen species(ROS), inducible isoforms of NO synthase(iNOS), cyclooxygenase-2(COX-2) to investigate the anti-inflammatory effect of LOF extracts. In addition, we measured generation of pro-inflammatory cytokines(TNF-${\alpha}$, IL-6). Results Cell viability showed that LOF extracts had no cytotoxicity in Raw 264.7 cells. The treatment with LOF extracts significantly decreased the generation of NO and pro-inflammatory cytokines(TNF-${\alpha}$, IL-6) in LPS-stimulated macrophage cells. Furthermore LOF extracts inhibited intracellular ROS generation dose dependently and reduced the expression of iNOS, COX-2 proteins. Conclusions These results showed that the LOF extracts had an anti-inflammatory effect on LPS-stimulated Raw 264.7 cells. These findings provide scientific support for the use of this Ligustrum obtusifolium S. for inflammatory-related diseases.

• 한국동물학회지
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• 제36권3호
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• pp.433-438
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• 1993

본 연구를 통해서 근세포 융합과정에서 신호 전달물질의 가능성이 제기되고 있는 세포내 cGMP peak가 guanylate cyclase activity의 변화와 관련이 있으며, guanylate cyclase는 L-arginine: NO synthase에 의해서 촉진될 것임을 입증할 수 있는 간접적 증거를 제시한다. 즉, SNP는 근세포의 융합과 guanylate cyclase activity를 아울러 증가시키며, L-arginine: NO synthase inhibitor인 L-NG-monomethyl arginine은 biochemical differentiation에는 영향을 주지 않고 근세포 융합만을 억제한다. 이러한 결과들과 muthylene blue가 근세포의 융합만을 억제하면서도 biochemical differentiation에는 영향을 주지 않으며 guanylate cyclase activity를 억제하는 사실들을 종합해서 생각할 때, 근세포 융합에서 cGMP peak가 guanylate cyclase activity의 활성화와 관련이 있으며, L-arginine: NO synthase가 $Ca^2$+ influx와 guanylate cyclase 사이를 매개할 가능성을 암시한다.

• Natural Product Sciences
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• 제24권3호
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• pp.181-188
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• 2018

Caryopteris incana (Verbenaceae) has been used to treat cough, arthritis, and eczema in Oriental medicine. The two fractions ($CHCl_3-$ and BuOH fractions) and the essential oil of the plant material were subjected to the inducible nitric oxide synthase (iNOS) assay. The $IC_{50}$ of the $CHCl_3$ fraction and the essential oil on LPS-induced macrophage RAW 264.7 cells were $16.4{\mu}g/mL$ and $23.08{\mu}g/mL$, respectively. On gas chromatography (GC)-mass spectroscopy (MS) analysis, twenty-five components representing 85.5% amount of total essential oil were identified. On the chromatogram, three main substances, trans-pinocarveol, cis-citral, and pinocarvone, occupied 18.8%, 13.5% and 18.37% of total peak area. Furthermore, by HPLC-UV analysis, six compounds including one iridoid (8-O-acetylharpagide)- and five phenylethanoid glycosides (caryopteroside, acteoside, phlinoside A, 6-O-caffeoylphlinoside, and leucosceptoside A) isolated from the BuOH fraction were quantified. The content of six compounds were shown as the following order: caryopteroside (162.35 mg/g) > 8-O-acetylharpagide (93.28 mg/g) > 6-O-caffeoylphlinoside (28.15 mg/g) > phlinoside (22.60 mg/g) > leucosceptoside A (16.87 mg) > acteoside (7.05 mg/g).

• Molecules and Cells
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• 제26권1호
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• pp.48-52
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• 2008

We here demonstrate an anti-inflammatory action of raloxifene, a selective estrogen receptor modulator, in lipopolysaccharide (LPS)-induced murine macrophage RAW264.7 cells. Treatment with raloxifene at micromolar concentrations suppressed the production of nitric oxide (NO) by down-regulating expression of the inducible nitric oxide synthase (iNOS) gene in LPS-activated cells. The decreased expression of iNOS and subsequent reduction of NO were due to inhibition of nuclear translocation of transcription factor NF-${\kappa}B$. These effects were significantly inhibited by exposure to the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor, LY294002, or by expression of a dominant negative mutant of PI 3-kinase. In addition, pretreatment with raloxifene reduced LPS-induced Akt phosphorylation as well as NF-${\kappa}B$ DNA binding activity and NF-${\kappa}B$-dependent reporter gene activity. Thus our findings indicate that raloxifene exerts its anti-inflammatory action in LPS-stimulated macrophages by blocking the PI 3-kinase-Akt-NF-${\kappa}B$ signaling cascade, and eventually reduces expression of pro-inflammatory genes such as iNOS.

• Advances in Traditional Medicine
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• 제10권3호
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• pp.155-164
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• 2010

Lycii fructus is the fruit of Lycium chinense Miller and is part of the Solanaceae family. Lycii fructus produces various effects such as hypotensive, hypoglycemic, anti-pyretic, and anti-stress activities. Lycii fructus is known to contain betaine, carotene, nicotinic acid, zeaxanthin, and cerebroside. In the present study, the effects of Lycii fructus aqueous extract on lipopolysaccharide (LPS)-induced inflammation in murine raw 264.7 macrophage cells were investigated. In this study we utilized the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcriptionpolymerase chain reaction (RT-PCR), Western blotting, and nitric oxide (NO) detection. Lycii fructus aqueous extract suppressed NO production by inhibiting the LPS-induced expressions of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-$\alpha$) mRNA and iNOS protein in murine raw 264.7 macrophage cells. Also, Lycii fructus aqueous extract suppressed the activation of nuclear factor-kappa B (NF-${\kappa}B$) in the nucleus. These results demonstrated that Lycii fructus aqueous extract causes an anti-inflammatory effect that was likely produced by the suppression of iNOS expression through the down-regulation of NF-$\hat{e}B$ binding activity.

• Toxicological Research
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• 제13권3호
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• pp.215-222
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• 1997

The features of seizure-related brain injuries in rats poisoned i.p. with diisopropylfiuorophosphate were investigated. Pyridostigmine bromide (0.1 mg/kg) and atropine methylnitrate (20 mg/kg), which are centrally inactive, were pretreated i.m. 30 min and 10 min, respectively. before diisopropylfluorophosphate (10 mg /kg, $2LD_50$) poisoning to reduce the mortality and eliminate peripheral signs. Diisopropylfluorophosphate induced severe limbic seizures, and early necrotic and delayed apoptotic brain injuries. The necrotic brain injury, which was closely related to seizure intensity, was exerted as early as 1 hr predominently in hippocampus and piriform cortex. showing spongiform change (malacia) of neurophils in severe cases, in contrast to a typical apoptotic (TUNEL-positive)pattern after 12 hr in thalamus, and a mixed type in amygdala. Nitric oxide content in cerebrospinal fiuid significantly increased after 2 hr, reaching a maximal level at 6 hr. Pretreatment with $_L-N^G$-nitroarginine, an inhibitor of nitric oxide synthase, reduced nitric oxide content and attenuated only apoptotic brain injury in all four brain regions examined without affecting seizure intensity and necrotic injury. Taken together, early necrotic and delayed apoptotic brain injuries induced by diisopropylfiuorophosphate poisoning in rats may be related to seizure intensity and nitric oxide production, respectively.

• Archives of Pharmacal Research
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• 제27권1호
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• pp.94-98
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• 2004

Effects of dopaminergic drugs on the degranulation of mast cells (RBL-2H3 cells) and the nitric oxide production from macrophage cells (RAW 264.7) were studied. Among the dopaminergic agonists and antagonists tested, bromocriptine, 7-OH-DPAT, haloperidol, and clozapine showed potent inhibitions of mast cell degranualtion ($IC_{50} value, 5 \mu$ M). However, these dopaminergic agents did not affect the tyrosine phosphorylations of the signaling components of the high affinity IgE receptor ($Fc\varepsilonRI$), such as Syk, $PLC\gamma1$, and $PLC\gamma2$.; This suggested that these signaling components were not involved in the inhibition of the mast cell degranulation by these compounds. On the other hand, dopamine, bromocriptine, 7-OH-DAPT, and haloperidol markedly inhibited the nitric oxide production from RAW 264.7 cells ($IC_{50}$ values, 10-20$\mu$M). Bromocriptine, a dopamine agonist that is routinely used for the treatment of Parkinsons disease, inhibited the expression of the inducible nitric oxide synthase at an early stage of the LPS-induced protein expression in a dose-dependent manner. The results suggested that these dopaminergic agents, when used for the treatment of dopamine receptors-related diseases, such as Schizophrenia or Parkinsons disease, might have additional beneficial effects.

• Natural Product Sciences
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• 제11권2호
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• pp.85-88
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• 2005

The methanolic extract from the seeds of Arctium lappa was found to inhibit the LPS-induced nitric oxide (NO) production in murine macrophage RAW264.7 cells. Bioassay-guided fractionation of a methylene chloride soluble fraction led to the isolation of three lignan compounds, arctiin (1), arctigenin (2), and lappaol B (3). Their structures were elucidated by UV, IR, MS, and NMR data, as well as by comparison with those of the literatures. Arctigenin (2) and lappaol B (3) had an iNOS inhibitory activity with $IC_{50}$ values of 12.5 and $25.9\;{\mu}M$, respectively.

• Toxicological Research
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• 제18권2호
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• pp.117-127
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• 2002

Saururus chinensis Baill has been used in Korean folk medicine for the treatment of various diseases such as edema, Jaundice, and furuncle. The components of this plant were extracted into four fraction. Among the four fraction, hexane and ethyl acetate fraction were highly toxic to 3T3 mouse embryo fibroblast and Raw 264.7 mouse macrophage, but n-butanol and residue fraction did not show any toxic effect to those cell lines. n-Butanol and residue fraction exhibited antioxidant effects on hydro-gen peroxide, hydroxyl radical, and superoxide anion directly in vitro and in the 3T3 fibroblasts. All the four fractions inhibited lipid peroxidation measured by thiobarbituric acid reactive substances (TBARS) formation. In addition, n-butanol and residue fraction showed inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide production, and also down-regulated inducible nitric oxide synthase (iNOS) mRNA transcription 6 h after LPS stimulation in Raw 204.7 cells. Only n-butanol fraction, which mainly consists of flavonoids, inhibited NF-kB activation by decreasing IkBa degradation 90 min after LPS stimulation. horn the results, it is suggested that this plant could be a good candidate material for drug development based on its antioxidant and/or anti-inflammatory constituents.

• 대한화장품학회지
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• 제32권2호
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• pp.111-116
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• 2006

Essential oils은 식물의 2차 대사산물로서 항박테리아, 항바이러스, 해열 및 항염증작용 등 다양한 생리활성을 가지고 있어 화장품 및 아로마테라피 소재를 비롯하여 식품, 항료, 의약품 원료 등 다양한 용도로 사용되고 있다. 뿐만 아니라, 최근 에스테틱 분야에서도 염증성 피부트러블개선, 주름개선, 노화방지 등 다양한 효능의 피부미용 개선을 목적으로 한 피부관리요법이나 미용관련제품소재로 널리 사용되고 있으나 이에 대한 체계적/객관적 효능 검증 및 작용기작 규명에 대한 과학적 연구는 매우 미비한 실정이다. 따라서, 본 연구는 피부염증과 연관지어 33종의 essential oils의 항염증 기능을 알아보기 위하여 중요 염증 매개인자의 일종인 nitric oxide (NO)를 탐색하였다. 그 중 NO 생성을 효과적으로 억제시키는 essential oil을 선택하여 농도별로 NO 생성억제효과, 세포독성, inducible nitric oxide synthase (iNOS) 발현에 미치는 영향을 탐색하고 작용기작을 규명하고자 하였다. medical skin care에서 주로 사용되는 33종의 essential oils(각 $100 {\mu}g/mL$)의 NO 생성억제효과를 탐색한 결과, 항염증 효과로 알려진 Rosemery (12.3%), Chamomile (37.8%), Lavender (14.2%), Cedar Wood (17.3%), Cypress (7.5%) 보다 Lemongrass (95.3%)의 NO 생성 억제율이 높았으며, iNOS발현에서도 억제 효과를 보여주었다. Lemongrass는 농도의존적으로 NO 생성을 억제시켰으며, $IC_{50}$값은 $22 {\mu}g/mL$이다. 결론적으로 Lemongrass oil은 체내 염증반응의 중요 매개인자인 NO의 생성을 억제하는 항염증 물질로서 염증성 피부질환인 여드름피부의 개선 및 예방 제품 개발에 좋은 소재로서의 가능성이 제시되어진다.