• 대한본초학회지
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• 제23권2호
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• pp.179-192
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• 2008

Objectives : Draconis Resina (DR) has been used as a traditional Korean herbal medicine since ancient times, and today it is used as a medication for wounds, tumors, diarrhea, rheumatism, in the itching of insect bites and with other conditions in the folk medicine. The aim of this study was to determine whether fractionated extracts of DR inhibit free radical generation, intracellular oxidation, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, Methods : DR extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of DR onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H202, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 formation in macrophages. Conclusions : Our results indicate that dichloromethane and ethyl acetate extracts of DR have potential as an agent of chronic inflammatory diseases.

• Journal of Ginseng Research
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• 제39권2호
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• pp.162-168
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• 2015

Background: Although the aerial parts of hydroponic Panax ginseng are reported to contain higher contents of total ginsenosides than those of roots, the isolation and identification of active metabolites from the aerial parts of hydroponic P. ginseng have not been carried out so far. Methods: The aerial parts of hydroponic P. ginseng were applied on repeated silica gel and octadecylsilane columns to yield four glycosyl glycerides (Compounds 1-4), which were identified based on nuclear magnetic resonance, infrared, fast atom bombardment mass spectrometry, and gas chromatography/mass spectrometry data. Compounds 1-4 were evaluated for inhibition activity on NO production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Results and conclusion: The glycosyl glycerides were identified to be (2S)-1-O-7(Z),10(Z),13(Z)-hexadecatrienoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (1), (2S)-1-O-linolenoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (2), (2S)-1-O-linolenoyl-2-O-linolenoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (3), and 2(S)-1-O-linoleoyl-2-O-linoleoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (4). Compounds 1 and 2 showed moderate inhibition activity on NO production in LPS-stimulated RAW264.7 cells [half maximal inhibitory concentration ($IC_{50}$): $63.8{\pm}6.4{\mu}M$ and $59.4{\pm}6.8{\mu}M$, respectively] without cytotoxicity at concentrations < $100{\mu}M$, whereas Compounds 3 and 4 showed good inhibition effect ($IC_{50}$: $7.7{\pm}0.6{\mu}M$ and $8.0{\pm}0.9{\mu}M$, respectively) without cytotoxicity at concentrations < $20{\mu}M$. All isolated compounds showed reduced messenger RNA (mRNA) expression of interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, and tumor necrosis factor-${\alpha}$ in LPS-induced macrophage cells with strong inhibition of mRNA activity observed for Compounds 3 and 4.

• 한국식품영양과학회지
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• 제44권1호
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• pp.7-13
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• 2015

본 연구에서는 아로니아 열매 추출물(AF-H)의 항염증 활성을 조사하기 위하여 LPS 자극에 의해 유도된 RAW 264.7 macrophage의 염증반응에서 AF-H의 염증매개인자 및 염증성 사이토카인 분비 억제활성과 이에 관련된 세포 내 작용기전 해석을 수행하였다. LPS($1{\mu}g/mL$)로 RAW 264.7 세포를 24시간 자극하는 염증모델에서 세포독성을 나타내지 않는 안전한 농도의 AF-H($0{\sim}500{\mu}g/mL$)를 LPS 처리 12시간 전에 처리하여 NO 및 PGE2의 분비 억제활성을 측정하였다. 그 결과 AF-H 처리에 의해 NO와 PGE2의 생성이 처리 농도에 의존하여 유의하게 억제되었으며, 이들 염증매개인자의 생합성 효소인 iNOS 및 COX-2의 세포 내 발현도 현저하게 억제되는 것으로 관찰되었다. 또한 AF-H의 처리에 의해 염증성 사이토카인인 $TNF-{\alpha}$와 IL-6의 분비도 유의하게 억제되는 것으로 확인하였다. 이러한 AF-H에 의한 항염증 활성의 세포 내 기전을 해석하기 위하여 LPS 자극에 의해 유도되는 MAPK와 $NF-{\kappa}B$ 전사인자의 활성화에 대한 억제 효과를 조사하였다. 그 결과 AF-H는 MAPK의 인산화에는 별다른 영향을 미치지 않고 $NF-{\kappa}B$의 활성화($I{\kappa}B$ 인산화)를 효과적으로 억제하는 것으로 확인되었다. 한편 LPS에 의한 in vivo 패혈증 모델에서 AF-H에 의한 패혈증 억제활성을 측정한 결과 비록 통계학적으로 유의하지는 않으나 AF-H 투여에 의해 생존율과 50% 사망률의 연장 효과가 관찰되었다. 이들 결과를 종합해 보면 아로니아 열매 열수추출물은 $NF-{\kappa}B$의 활성화 억제를 통해 NO, PGE2, $TNF-{\alpha}$ 및 IL-6 등의 염증매개인자와 사이토카인의 생성을 억제하는 항염증 활성을 지니는 것으로 확인되었다.

• 한국자원식물학회지
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• 제28권5호
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• pp.574-581
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• 2015

본 연구는 누은분홍잎(Acrosorium yendoi Yamada) 80% 에탄올를 가지고 추출한 후 추출물을 극성에 따라 순차적으로 용매분획을 실시하여, 80% 에탄올 추출물 및 그 분획물들의 염증반응의 주체가 되는 대식세포 계열인 RAW 264.7 세포에서 LPS로 유도된 NO의 생성억제 효과, 그리고 iNOS와 COX-2의 단백질 발현 억제효과, PGE₂ 생성 억제 효과 및 TNF-α와 IL-6등과 같은 pro-inflammatory cytokines 생성 억제효과 등을 알아보았다. 대식세포 계열인 RAW 264.7 세포에 LPS로 자극을 주고 누은분홍잎 80% 에탄올 추출물 및 분획물을 처리하여 확인해본 결과, 에틸아세테이트 분획물에서 NO와 PGE₂ 생성 억제 활성이 가장강하게 나타났으며, 농도 의존적으로 NO 및 PGE₂ 생성억제 활성을 보임을 확인할 수 있었고, 세포독성평가(LDH)에서는 누은분홍잎 80% 에탄올 추출물 및 분획물에 50㎍/㎖ 이하의 농도에서는 세포독성이 나타나지 않았다. 또한, 누은분홍잎의 80% 에탄올 추출물, 헥산 및 에틸아세테이트 분획물에서 iNOS 및 COX-2 발현 억제 활성을 보이고 있음을 확인할 수 있었으며, 에틸아세테이트 분획물의 경우는 뚜렷하게 농도 의존적으로 iNOS 및 COX-2 발현 억제 활성을 보임을 알 수 있었다. 이상의 결과는 누은분홍잎에 대한 효능, 이화학적 성분 및 유효성분등에 대한 연구가 전무하여 본 결과를 통해 누은분홍잎 추출물 및 활성 분회물을 이용한 항염증 효능을 갖는 유효성분 분리 및 이를 통한 작용기전 연구에 중요한 기초자료가 될 것이라 사료되고, 또한 안전하고 효과적인 항염증 관련 의약품, 의약외품 및 기능성 식품소재 개발에 대한 가능성을 제시하였다.

• 한국약용작물학회지
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• 제24권3호
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• pp.228-236
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• 2016

Background: Sedum takesimense Nakai has been used as folk medicine in Korea. The present study aimed to determine the biological activity of S. takesimense by investigating the anti-inflammatory effects of S. takesimense water extract (SKLC) on the lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Methods and Results: Cytotoxicity of SKLC on RAW 264.7 cells was determinded by performing MTS assay was found to have no cytotoxic effect on RAW 264.7 cells at a concentration range of $62-500{\mu}g/m{\ell}$. Further, pretreatment of SKLC inhibited lipopolysaccharide-induced nitric oxide (NO) production in a dose-dependent manner. To determined the inhibitory mechanisms of SKLC on inflammatory mediators, we assessed the inducible nitric oxide synthase (iNOS) and cyclooxygnease-2 (COX-2) pathways. The activities of these pathways were decreased in a dose-dependent manner by SKLC. The production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6 were also reduced. Conclusions: These results suggest that the down regulation of iNOS, COX-2, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 expression by SKLC are mediated by the down regulation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity, a transcription factor necessary for pro-inflammatory mediators. This might be the mechanism underlying the anti-inflammatory effects of SKLC.

• 생명과학회지
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• 제27권1호
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• pp.8-14
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• 2017

치자 열매는 아시아에서 음식과 옷의 염료로 사용되어 왔다. 본 연구에서는 BV-2 소교세포에서 치자열매 열수추출물(GJ)의 항염증 효과를 관찰하고 그 작용 기전을 연구하였다. GJ는 세포에 독성을 유도하지 않으면서 lipopolysaccharide로 인한 nitric oxide (NO) 분비와 inducible nitric oxide synthase (iNOS) 생성 및 활성산소 생성을 억제하였다. 또한 GJ는 농도의존적으로 heme oxygenase-1 (HO-1)의 발현을 유도하였다. 더군다나 HO-1 siRNA를 처리했을 때는 GJ가 iNOS의 발현을 억제하지 못하였다. GJ는 HO-1의 발현에 관여하는 전사인자인 nuclear factor E2-related factor 2를 핵으로 이동시켰다. 또한 GJ에 의한 HO-1의 발현은 phosphatidylinositol 3-kinase(PI-3K) 및 p38 kinase 억제제에 의해 감소되었으며, GJ가 Akt와 p38 kinase의 인산화를 유도하였다. 이상의 결과를 종합해보면, GJ는 PI3K/Akt 및 p38 신호전달과정을 통해 HO-1의 발현을 유도함으로써 NO와 같은 염증매개물질의 생성을 억제한다는 것을 알 수 있다. 이러한 연구결과는 치자열매가 신경염증을 억제하는 새로운 기전을 밝힌 것이다.

• 대한한방내과학회지
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• 제30권4호
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• pp.788-798
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• 2009

목적 : 이 연구는 백련초의 천식마우스 모델에 대한 효과를 조사하기 위해 수행하였다. 방법: 폐 세척액의 전체 세포와 호산구 수 그리고 폐조직으로의 면역세포의 침윤은 헤마톡실린과 에오신 염색으로 관찰 하였다. Interleukin (IL)-4의 발현량은 Enzyme-linked immunosorbent assay을 이용하여 측정하였으며, NO는 Griess roagont을 이용하였다. 결과 : 천식 병리현상의 하나인 Ovalbumin(OVA)에 의한 전체 세포와 호산구 수의 증가는 백련초 추출물 (200 mg/kg)을 처리한 군에서 감소하였다 (respectively, p<0.001 and p<0.01). 또한 OVA에 의한 증가된 면역세포의 침투는 백련초 추출물에 의해 감소하였다 (p < 0.01). 호산구의 침윤증상은 interleukin (IL)-4 그리고 5를 포함한 T helper (Th) 2 반응과 관련있기 때문에 폐세척액에서 IL-4의 발현량을 조사하였다. OVA에 증가한 IL-4는 백련초 추출물에 의해 감소하였다 (p < 0.05). 천식이 염증성 질병이기 때문에, lipopolysaccharide (LPS)로 자극한 RAW 264.7 세포에서 nitrite (NO) 생성을 살펴보았다. LPS에 의해 증가한 NO 생성을 백련초 추출물 (50, 100 and 200 mg/ml)에서 감소하였다 (respectively, p < 0.05, p<0.01 and p<0.05). 결론 : 이런 결과들은 백련초가 천식의 호산구 침윤에 저해효과를 가지고 있다고 나타내며, 백련초 추출물의 천식을 포함한 염증성 질환의 치료에 치료후보물질로 제안할 수 있다.

• 대한간호학회지
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• 제23권4호
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• pp.528-543
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• 1993

Vitamin E, which has its advocates in the treatment of diabetes mellitus. autoimmune disease, cancer and peripheral vascular and thromboembolic disease, has now been alleged to have a powerful antioxident effect and to affect various biological activities such as fertility factor, inhibition of human platelet aggregation and stabilization of biological membranes. The present study was designed to test whether vitamin I(alpha-tocopherol) can : (1) enhance the hemagglutinin response to sheep red blood cells (SRBC), (2) modulate Arthus and delayed type hypersensitivity(DTH) to SRBC and contact hypersensitivity to dinitrofluorobenzene (DNFB). (3) enhance the mitogenic response of murine splenocyte, (4) decrease the recovery of Cryptococcus neoformans from brain, lung, liver, spleen and kidney of infected mice and (5) have an inhibitory or enhancing effect on the induction of active systemic anaphylaxis(ASA) induced by chicken-gamma globulin (CGG) in mice. Mice were given either intramuscular injections of 0.3ml (300mg) of vitamin I before immunization or were infection for 10 consecutive days or were given by vitamin I esophageal intubation, 0.1ml(100mg), for 20 days before sacrifice for the mitogenic response experiments. It was found that vitamin E treated mice showed a significant enhancement in hemagglutinin response, Arthus reaction and DTH to SRBC and contact hypersensitivity to DNFB. There was no significant difference in the mitogenic response to phytohemagglutinin(PHA), but the response to concanavalin A(ConA) or pokeweed mitogem(PWM) was increased in vitamin E-treated mice. Interestingly, the vitamin E administration before C. neoformans infection decreased significantly the recovery of C. neoformans from brain lung, liver, spleen and kidney of the infected mice as compared with that of the control mice, strongly suggesting that vitamin E pretreatment may increase the resistance of mice to the fungal infection. Unexpectedly, vitamin E administration enhanced the production of CGG -induced ASA. Taken together, it can be concluded that vitamin I administration may in-crease the humoral and cellular immune response and resistance. to C. neoformans infection, but enhance the induction of ASA to CGG. Further studies are necessary to clarify the underlying mechanism accounting for these effects.

• Animal Bioscience
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• 제34권11호
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• pp.1766-1775
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• 2021

Objective: The oxidative stress status and changes of chicken ovary tissue after shading were studied, to determine the mechanism of the effect of shading on follicular development. Methods: Twenty healthy laying hens (40 weeks old) with uniform body weight and the same laying rate were randomly divided into two groups (the shading group and normal light group). In the shading group, the cage was covered to reduce the light intensity inside the cage to 0 without affecting ventilation or food intake. The normal lighting group received no additional treatment. After 7 days of shading, oxidative stress related indicators and gene expression were detected. Results: Analysis of paraffin and ultrathin sections showed that apoptosis of ovarian granulosa cells (GCs) increased significantly after light shading. Enzyme linked immunosorbent assay results revealed that the levels of total antioxidant capacity, malondialdehyde, superoxide dismutase (SOD), glutathione, catalase (CAT), and other substances in the sera, livers, ovaries, and follicular GCs of laying hens increased significantly after shading for 7 days; and reactive oxygen species (ROS) levels in the livers of laying hens also increased significantly. ROS in the serum, ovarian and GCs also increased. After shading for 7 days, the levels of 8-hydroxy-2 deoxyguanosine in the sera and ovarian tissues of laying hens increased significantly. Cell counting kit-8 detection showed that the proliferation activity of GCs in layer follicles decreased after shading for 7 days; the expression level of the anti-apoptotic gene B-cell lymphoma-2 in ovarian tissue and follicular GCs was significantly reduced, and the expression levels of pro-apoptotic caspase 3 (casp3), and SOD, glutathione peroxidase 2 (GPX2), and CAT were all significantly increased. Conclusion: Oxidative stress induced by shading light has a serious inhibitory effect on follicular development during reproduction in laying hens.

• ALGAE
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• 제29권4호
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• pp.333-341
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• 2014

In this study, four compounds isolated from the red alga Laurencia snackeyi were evaluated for their potential anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. These compounds were tested for their inhibitory effects on nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. Since $5{\beta}$-hydroxypalisadin B showed the best activity it was further tested for the production of prostaglandin-$E_2$ ($PGE_2$), expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the release of pro-inflammatory cytokines tumor necrotic factor-alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6). $5{\beta}$-Hydroxypalisadin B significantly reduced the $PGE_2$ release and suppressed the iNOS and COX-2 expression in LPS-stimulated RAW 264.7 cells. It also significantly reduced the release of pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These findings provide the first evidence of anti-inflammatory potential of $5{\beta}$-hydroxypalisadin B isolated from the red alga L. snackeyi and hence, it could be exploited as an active ingredient in pharmaceutical, nutraceutical and functional food applications.