• Biomolecules & Therapeutics
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• 제24권5호
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• pp.543-551
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• 2016

This study was designed to evaluate the pharmacological effects of Vaccinium bracteatum Thunb. methanol extract (VBME) on microglial activation and to identify the underlying mechanisms of action of these effects. The anti-inflammatory properties of VBME were studied using lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. We measured the production of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin $E_2$ ($PGE_2$), tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-1 beta (IL-$1{\beta}$), and interleukin-6 (IL-6) as inflammatory parameters. We also examined the effect of VBME on intracellular reactive oxygen species (ROS) production and the activity of nuclear factor-kappa B p65 (NF-${\kappa}B$ p65). VBME significantly inhibited LPS-induced production of NO and $PGE_2$ and LPS-mediated upregulation of iNOS and COX-2 expression in a dose-dependent manner; importantly, VBME was not cytotoxic. VBME also significantly reduced the generation of the pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. In addition, VBME significantly dampened intracellular ROS production and suppressed NF-${\kappa}B$ p65 translocation by blocking $I{\kappa}B-{\alpha}$ phosphorylation and degradation in LPS-stimulated BV2 cells. Our findings indicate that VBME inhibits the production of inflammatory mediators in BV-2 microglial cells by suppressing NF-${\kappa}B$ signaling. Thus, VBME may be useful in the treatment of neurodegenerative diseases due to its ability to inhibit inflammatory mediator production in activated BV-2 microglial cells.

• IMMUNE NETWORK
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• 제6권1호
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• pp.27-32
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• 2006

Background: Chronic inflammation in the brain has known to be associated with the development of a various neurological diseases including dementia. In general, the characteristic of neuro-inflammation is the activated microglia over the brain where the pathogenesis occurs. Pro-inflammatory repertoires, interleukin-1${\beta}$ (IL-1${\beta}$) and nitric oxide (NO), are the main causes of neuro-degenerative disease, particularly in Alzheimer's disease (AD) which is caused by neuronal destruction. Those pro-inflammatory repertoires may lead the brain to chronic inflammatory status, and thus we hypothesized that chronic inflammation would be inhibited when pro-inflammatory repertoires are to be well controlled by inactivating the signal transduction associated with inflammation. Methods: In the present study, we examined whether biphenyl dimethyl dicarboxylate (DDB), an active compound from Schizandra chinensis Baillon, inhibits the NO production by a direct method using Griess reagent and by RT-PCR in the gene expression of inducible nitric oxide synthase (iNOS) and IL-1${\beta}$. Western blots were also used for the analysis of NF-${\kappa}B$ and I${\kappa}B$. Results: In the study, we found that DDB effectively inhibited IL-1${\beta}$ as well as NO production in BV-2 microglial cell, and the translocation of NF-${\kappa}B$ was comparably inhibited in the presence of DDB comparing those to the positive control, lipopolysaccharide. Conclusion: The data suggested that the DDB from Schizandra chinensis Baillon may play an effective role in inhibiting the pro-inflammatory repertoires which may cause neurodegeneration and the results imply that the compound suppresses a cue signal of the microglial activation which can induce the brain pathogenesis such as Alzheimer's disease.

• Asian-Australasian Journal of Animal Sciences
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• 제24권3호
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• pp.414-420
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• 2011

A study was conducted to investigate the effects of the combined stressor induced by high stocking density with feed restriction on immunological parameters such as leukocyte differential counts and cytokine expression in laying hens. A total of forty White Leghorn laying hens were randomly allotted into the control (12 kg of body weight/$m^2$) and the stress (44 kg of body weight/$m^2$) groups, and then birds of the stress group were given 75% of voluntary intake of the control birds for 12-d on a daily basis. There was a significant decrease in body weight without affecting the relative weights of the liver and spleen after 12-d of the combined stressor. In hematological values, no significant difference in leukocyte differential counts including heterophils (H), lymphocytes (L), monocytes and H:L ratio was observed between the two groups. In cytokines, hepatic lipopolysaccharide-induced tumor necrosis factor-${\alpha}$ (LITNF-${\alpha}$) and inducible nitric oxide synthase (iNOS) expression levels in the stress group were significantly (p<0.05) higher compared with those in the control group. However, the expression levels of interleukin (IL)-4 and IL-6 in the liver were not affected by the combined stressor. Splenic LITNF-${\alpha}$ expression in the combined stressor group was significantly (p<0.05) up-regulated compared with that in the control birds. However, the combined stressor did not affect splenic IL-4, IL-6 and iNOS expression. In conclusion, the combined stressor caused by high stocking density with feed restriction enhanced some pro-inflammatory cytokines including LITNF-${\alpha}$ and iNOS in lymphoid and non-lymphoid organs of birds, suggesting that altered cytokine expression to given stressors can be another parameter that can be used in assessing stress responses of birds.

• Tuberculosis and Respiratory Diseases
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• 제52권6호
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• pp.616-626
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• 2002

연구배경 : PM2.5는 호흡기질환의 악화 및 호흡기 질환에 의한 사망률과 밀접하게 관계가 있다. PM2.5에 의한 호흡기 염증반응의 악화가 그 원인으로 생각되나 염증반응의 조절기전은 확실하지 않다. 본연구에서는 PM2.5에 의한 폐장의 염증반응을 조절하는데 중요한 역할을 하는 전사인자인 $NF{\kappa}B$의 활성화 기전을 알아보아 염증 조절기전에 대한 이해를 높이고자 시도되었다. 방 법 : 폐상피세포주인 A549 세포에서 PM2.5 자극에 의한 $NF{\kappa}B$의 활성화 및 ROS, 그리고 RNS 분비를 관찰 하였다. iNOS 억제제인 L-NIL가 $NF{\kappa}B$ 활성화에 미치는 영향을 관찰하였다. 결 과 : PM2.5는 A549 세포에서 $NF{\kappa}B$의 활성화를 유도 하였다. PM2.5로 A549 세포를 자극시 즉각적인 RNS의 분비는 관찰되었으나 명확한 ROS의 증가는 관찰되지 않았다. RNS 억제제인 L-NIL 처리시 $NF{\kappa}B$ 활성화는 억제되었다. 결 론 : A549 세포에서 PM2.5에 의하여 유도된 즉각적인 $NF{\kappa}B$ 활성화 과정에는 RNS가 중요한 역할을 할 것으로 생각된다.

• Tuberculosis and Respiratory Diseases
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• 제62권4호
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• pp.308-313
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• 2007

연구배경: 패혈증에서 Nitric oxide 스테로이드 호르몬은 혈역학적 변화와 염증반응에 관여하는데 이 두 인자는 서로 상관성이 있는 것으로 알려지고 있다. 하지만 실제 환자에서 서로의 상관성이나 임상적 의의는 연구가 부족한 실정이다. 방법: 패혈증 환자 26예와 대조군 14예를 대상으로 혈중 총 NO와 혈중 코티졸 농도를 측정하였고 이어서 제 3, 5, 7병일에도 연속적으로 측정을 하였다. 결과: 패혈증 환자군에서 초기 혈중 코티졸 및 총 NO 농도는 대조군에 비하여 유의하게 증가하였고 경증 패혈증에 비하여 중증 패혈증 환자군에서 유의하게 높았다. 초기 혈중 총 NO의 농도는 APACHE II 점수, 정맥혈 lactate 농도와 상관성이 있었다. 패혈증의 시간의 경과에 따라 혈중 NO 농도는 제 1병일, 제 5병일, 제 7병일에 혈중 코티졸의 농도와 유의한 상관성이 있었다. 결론: 패혈증 환자들에서 혈중 NO와 코티졸 농도는 증가되어 있었으며, 경과에 따라 서로 유의한 상관성이 지속되었다. 상호작용기전에 대하여는 추가적인 연구가 필요할 것이다.

• 생명과학회지
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• 제22권9호
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• pp.1254-1260
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• 2012

모링가(Moringa oleifera Lam.)는 항알러지 약물로써 식용 가능한 식물이다. 본 연구에서 부위별 모링가의 피부 보호제로서의 가능성을 확인하기 위하여, TNF-${\alpha}$로 염증을 유도한 각질형성세포에서 모링가의 씨, 뿌리, 잎과 열매 메탄올 추출물의 항염증 효과를 비교 실험하였다. 피부세포의 콜라겐 분해 관련인자인 MMP-2, MMP-9의 효소 활성을 측정한 결과 모든 부위별 모링가 추출물이 MMP-9의 효소 활성을 감소시켰다. 특히 모링가 뿌리 추출물은 낮은 농도에도 MMP-9을 효과적으로 감소시켰으며 MMP-2의 효소활성 억제에도 효과가 관찰되었다. 또한 피부 염증관련 인자로 알려진 iNOS와 COX-2의 단백질 발현을 측정한 결과, COX-2의 단백질 발현은 모링가 잎을 제외한 뿌리, 씨앗, 열매 추출물에 의해 억제되었다. 그 중 모링가 뿌리 추출물은 낮은 농도에서도 COX-2 단백질의 발현을 억제시켰다. 그러나 iNOS는 부위별 모링가 추출물에 의한 단백질 발현의 변화가 없는 것으로 나타났다. 뿐만 아니라 피부 염증을 일으키는 cytokine으로 알려진 IL-6의 mRNA발현을 확인한 결과 TNF-${\alpha}$에 의해 증가된 IL-6 발현을 모링가 뿌리 추출물이 효과적으로 억제 시키는 것을 확인할 수 있었다. 이상의 결과로 미루어 보아 부위별 모링가 추출물 중 뿌리 추출물에서 가장 피부노화 억제와 항 염증 효과가 높을 것으로 사료되며, 식물 유래의 피부 보호제 제품 개발에 있어 유용한 원료로 사용될 수 있을 것으로 생각된다.

• 대한본초학회지
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• 제30권5호
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• pp.59-65
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• 2015

Objectives : Fermentation of herbs has been known to be helpful in improving the immune systems and protecting body against disease. The present study was conducted to evaluate anti-inflammatory effects of the fermentation extracts (FE) consisting of soybean, red ginseng andCitrus UnshiuPeel in lipopolysaccharide (LPS)-activated Raw264.7 cells.Methods : FE were prepared by the fermentation withBacillus Subtilisand then by extraction with ethanol (95%; prepared by the fermentation process). Cell viability was measured by MTT assay. Nitric oxide (NO) production was measured in culture media by Griess assay. The expression of nuclear factor (NF)-κB and inhibitory kappa B alpha (IκBα) was determined by Western blot.Results : LPS-induced production of NO and PGE₂was dose-dependently decreased by the treatment of FE in Raw264.7 cells. These suppressive effects of FE on NO and PGE₂production were related to the inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. FE inhibited LPS-induced production of pro-inflammatory cytokines, TNF-α, IL-6, and IL-1βin a dose-dependent manner. Furthermore, FE inhibited the NF-κB signaling pathway through the prevention of LPS-induced degradation of IκBαin cytosol and the nuclear translocation of NF-κB.Conclusions : These findings suggest that FE could have anti-inflammatory effects on LPS-induced inflammatory responses in macrophages.

• 대한한의학방제학회지
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• 제18권1호
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• pp.169-179
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• 2010

The pistil of nelumbo nucifera (PNN) is used in the treatment of nocturnal pollution, hematemesis, epistaxis, metrorrhagia and diarrhoea in traditional medicine. The present study was examined to evaluate the effects of PNN on the production of pro-inflammatory mediators in vitro. After the treatment of PNN, cell viability was measured by MTT assay, nitric oxide (NO) production was monitored by measuring the nitrite content in culture medium. The protein bands were determined by immunoblot analysis and levels of cytokines were analyzed by sandwich immunoassays. In the MTT assay, the doses of PNN extract (0.03, 0.10 mg/ml) had no significant cytotoxicity. The increases of NO production and inducible nitric oxide synthase expression were detected in lipopolysaccharide(LPS)-activated Raw 264.7 cells compared with control, in contrast, these increases were significantly attenuated by pre-treatment with PNN. In cytokine assay, the massive pro-inflammatory cytokines such as tumour necrosis factor-$\alpha$, interleukin (IL)-$1{\beta}$ and IL-6 were induced in LPS-activated Raw 264.7 cells, but pre-treatment of Raw 264.7 cells with PNN caused inhibition (TNF-$\alpha$=14.17%, IL-$1{\beta}$=107.43%, IL-6=46.27%) the production of cytokines by LPS. In addition, PNN reduced prostaglandin E2 productions in a dose-dependent manner (0.03mg/ml=37.52%, 0.10 mg/ml=83.77%) as a consequence of the inhibition of cyclooxygenase-2 expression. Taken together, our data indicates that PNN can regulate the inflammatory response in macrophage cells activated by Gram-negative infection.

• 한국약용작물학회지
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• 제24권1호
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• pp.31-37
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• 2016

Background : We studied the anti-oxidant activity and anti-inflammatory effects of Spiraea fritschiana Schneid extract (SFSE). Methods and Results : The SFSE was prepared using methanol and was evaluated for its total phenol and flavonoid content, DPPH (1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity, reducing power, and effect on nitric oxide (NO) production, and cell viability by using real-time polymerase chain reaction (PCR). The total phenol content was $212.78{\mu}g{\cdot}galli$c acid equivalent (GAE)/mg and the total flavonoid content was $66.84{\mu}g{\cdot}quercetin$ equivalent (QE)/mg. The extract showed antioxidant activity (DPPH free-radical scavenging activity) with $RC_{50}$ value of $76.61{\mu}g/m{\ell}$. The reducing power of the extract was Abs 0.58 at $250{\mu}g/m{\ell}$. Cell viability was determined using the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. To evaluate anti-inflammatory activity, we examined the inhibitory effects on lipopolysaccharide-(LPS)-induced NO production in RAW 264.7 cells. The NO inhibition rate was 90% at $200{\mu}g/m{\ell}$ SFSE. At the same concentration, the expression of pro-inflammatory genes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 also decreased. Conclusions : Our results suggest that SFSE is a novel resource for the development of foods and drugs that possess anti-oxidant and anti-inflammatory activity.

• 대한한방내과학회지
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• 제34권2호
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• pp.204-214
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• 2013

Objectives : This study was designed to investigate the cellular and molecular mechanisms of anti-inflammatory activity of the water extract of Polygala tenuifolia Willd. (Pt-WE). Methods : Using lipopolysaccharide (LPS)-stimulated murine RAW264.7 cells, we examined inflammatory mediators such as nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$). Also, the inhibitory effect of Pt-WE on the activity of activator protein 1 (AP-1) and upstream signaling molecules was evaluated. To assess the protective effect of Pt-WE on hydrochloride/ethanol (HCl/EtOH)-induced gastric ulcer in mice, we compared Pt-WE (200 mg/kg) with ranitidine (50 mg/kg) treated mice's gastric mucosa, based on gross observations. Results : Pt-WE inhibited LPS-induced production of NO, $PGE_2$ in a dose-dependent manner, without causing cytotoxicity. Pt-WE suppressed AP-1 activation by reducing generations of both c-Jun and c-Fos. In addition, Pt-WE inhibited the p-MKK 4/7 (mitogen-activated protein kinase kinase 4/7) and p-JNK (c-Jun N-terminal kinase) 1 in LPS-stimulated RAW264.7 cells. HCl/EtOH-induced gastric ulcer lesions were inhibited by pre-treatment of Pt-WE based on gross observations. In addition, Pt-WE decreased the phosphorylation level of JNK. Conclusions : These results demonstrate that Pt-WE has anti-inflammatory and gastroprotective effects. Thus, Pt-WE may be used widely in treatment of not only neurodegenerative diseases but also inflammatory diseases.