• 대한본초학회지
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• 제27권6호
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• pp.1-6
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• 2012

Objectives : ROS are involved in a wide spectrum of diseases including chronic inflammation and cancer. S.chinensis Baill, a perennial herb commonly called Chinese lizard's tail or Sam-baek-cho in Korea, is used for the treatment of edema and inflammatory diseases in the Oriental folk medicine. In this study, we investigated the antioxidant activities and anti-inflammatory effects of the two extracts, water(WE) and ethyl acetate(EAE) from S.chinensis Baill. Methods : Anti-oxidant activity was evaluated using Fe2+ chelating and hydroxyl radical scavenging assay. DNA cleavage assay, and western blot and immunostaining for phospho-p65 were performed to evaluate anti-oxidative effect. Anti-inflammatory effect was performed using NO generation assay and western blot in LPS-stimulated RAW264.7 cell. Results : In Fe2+ chelating activity and hydroxyl radical scavenging activity, WE showed more strong scavenging activity for hydroxyl radical than EAE. WE scavenged hydroxyl radical by 12% at 3.2 ${\mu}g/ml$, 21% at 16 ${\mu}g/ml$, 32% at 80 ${\mu}g/ml$, 66% at 400 ${\mu}g/ml$ and 82% at 2000 ${\mu}g/ml$, respectively. In addition, WE showed more strong chelating activity than EAE. WE chelated Fe2+ ion by 1.1% at 3.2 ${\mu}g/ml$, 8.2% at 16 ${\mu}g/ml$, 26.3% at 80 ${\mu}g/ml$, 72% at 400 ${\mu}g/ml$ and 89% at 2000 ${\mu}g/ml$, respectively. Also, WE inhibited oxidative damage via its anti-oxidant activity. In anti-inflammatory effect, EAE inhibited NO production and iNOS expression. In addition EAE suppressed the NF-${\kappa}B$ and MAPK signaling pathway in LPS-stimulated RAW 264.7 cells. Conclusions : Together, these data indicate that S. chinensis Baill, shows anti-oxidant activity and anti-inflammatory effect.

• 한국미생물·생명공학회지
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• 제43권2호
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• pp.112-119
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• 2015

본 연구에서는 큰잎모자반의 항염증 효과를 알아보기 위해 LPS에 의해 염증반응이 유도된 RAW 264.7 세포에 대한 큰잎모자반 에탄올 추출물의 항염증 효과를 살펴보았다. 세포내 염증매개성 cytokine (IL-6, TNF-α 및 IL-1β) 분비량의 측정결과, 농도 의존적으로 유의적 감소를 보였으며, 특히, 100 μg/ml로 처리하였을 경우 LPS 단독 처리와 비교 시 약 48% 이상의 높은 분비량 감소효과를 보였다. 추출물이 iNOS, COX-2 및 NF-kB 발현 억제에 미치는 효과를 알아본 결과, LPS 단독처리구에 의해 각 단백질의 발현량이 현저히 증가하였으나, 추출물을 처리하였을 때 그 발현량이 농도 의존적으로 감소하는 것을 확인할 수가 있었다. 특히 50 및 100 μg/ml 농도에서 PBS 처리구와 유사한 발현량을 보여 우수한 항염증 효과를 가짐을 확인하였다. 귀 부종 억제 효과 및 조직 관찰을 수행한 결과, 추출물 250 mg/kg 농도에서 prednisolone 10 mg/kg 처리와 유사하게 감소함을 보였다. 또한, croton oil로 부종을 유발한 마우스 귀 조직에서 croton oil만을 처리한 경우에 비해 추출물을 100 mg/ml 농도로 처리한 경우 prednisolone 처리구와 유사한 정도로 경피 및 진피 두께가 얇아진 것을 확인하였으며, 조직 내 mast cell 침윤을 현저히 억제함을 보였다. 이 결과를 종합해 볼 때, 큰잎모자반 에탄올 추출물이 염증 치료제로써의 소재로 이용될 가치가 충분할 것으로 사료된다.

• IMMUNE NETWORK
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• 제10권6호
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• pp.219-229
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• 2010

Background: N-myc downstream regulated gene 2 (NDRG2) is a member of the NDRG gene family. Our previous report indicated a possible role for NDRG2 in regulating the cytokine, interleukin-10 (IL-10), which is an important immunosuppressive cytokine. Several pathways, including p38-MAPK, NF-${\kappa}B$, and JAK/STAT, are used for IL-10 production, and the JAK/STAT pathway can be inhibited in a negative feedback loop by the inducible protein, SOCS3. In the present study, we investigated the effect of NDRG2 gene expression on IL-10 signaling pathway that is modulated via SOCS3 and STAT3. Methods: We generated NDRG2-overexpressing U937 cell line (U937-NDRG2) and treated the cells with PMA to investigate the role of NDRG2 in IL-10 production. U937 cells were also transfected with SOCS3- or NDRG2-specific siRNAs to examine whether the knockdown of SOCS3 or NDRG2 influenced IL-10 expression. Lastly, STAT3 and SOCS3 induction was measured to identify the signaling pathway that was associated with IL-10 production. Results: RT-PCR and ELISA assays showed that IL-10 was increased in U937-mock cells upon stimulation with PMA, but IL-10 was inhibited by overexpression NDRG2. After PMA treatment, STAT3 phosphorylation was decreased in a time-dependent manner in U937-mock cells, whereas it was maintained in U937-NDRG2 cells. SOCS3 was markedly reduced in U937-NDRG2 cells compared with U937-mock cells. IL-10 production after PMA stimulation was reduced in U937 cells when SOCS3 was inhibited, but this effect was less severe when NDRG2 was inhibited. Conclusion: NDRG2 expression modulates SOCS3 and STAT3 activity, eventually leading to the inhibition of IL-10 production.

• Journal of Nutrition and Health
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• 제41권3호
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• pp.224-231
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• 2008

본 연구는 십자화과 채소의 섭취로 체내유용 물질인 인돌이 전립선암 세포 PC3 cell의 항전이 효과 기전에 미치는 영향에 대하여 알아보았다. 인돌은 전립선암 세포중식을 농도 의존적으로 억제하였으며 인돌에 의한 세포 사멸의 영향과 관계없이 MMP-2, -9의 활성과 전사수준 및 단백질 발현을 억제하였다. 역으로 MMP활성 억제 물질인 TIMP-1,-2의 발현이 인돌 첨가에 의해 증가하였다. $NF{-\kappa}B$의 upstream에 존재하는 MAPK signaling 유전자인 ERK1/2, p38, JNK 발현이 인돌처리로 인산화를 억제하였다. 그리고 전립선암 세포 PC3 침윤성이 인돌 처리 시 유의적으로 감소하였다. 결론적으로 인돌은 PC3 인체 전립선암 세포의 전이 과정을 MAPK phthway를 통한 MMP 활성과 발현 억제, TIMP 발현 증가로 암 세포 전이 억제를 하는 것 으로 나타나 암 전이 억제 식품으로 가능성을 제시한다.

• 대한한방내과학회지
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• 제41권1호
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• pp.1-13
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• 2020

Objectives: Osteoarthritis (OA) is a chronic and degenerative joint disease characterized by progressive degeneration of articular cartilage. Inflammation is a recognized and important factor of OA progression. The present study was designed to investigate the protective effect of Corni Fructus water extract (CFW) on a monosodium iodoacetate (MIA)-induced rat model of OA. Methods: Osteoarthritis was induced by injection of MIA (50 µL; 80 mg/mL) into the knee joint cavity of rats. After an adaptation period for seven days, the rats were divided into 4 groups (n=8/group): normal, control, indomethacin-treated (5 mg/kg), and CFW-treated (200 mg/kg) groups. The rats were treated orally for 14 days. Pain was evaluated by determining hind paw weight distribution. For biochemical analyses, we measured the changes in reactive oxygen species (ROS) and peroxynitrite (ONOO^-) in the knee joint. The presence of anti-oxidant proteins and inflammatory proteins was determined by western blotting. Results: The administration of CFW significantly improved the hind paw weight distribution. The ROS and ONOO^- levels of knee joint were significantly decreased in the CFW group. CFW inhibited the production of inflammatory mediators, such as COX-2, and inflammatory cytokines, including IL-6 and IL-1β, via the NF-κB signaling pathway. The expression of anti-oxidant enzymes, such as catalase and GPx-1/2 also increased significantly. Conclusions: The findings indicate that CFW has a therapeutic and protective effect on OA by suppression of inflammation. Therefore, CFW could represent a potential and effective candidate for OA treatment.

• 대한본초학회지
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• 제34권1호
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• pp.117-124
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• 2019

Objective : Chronic acid reflux esophagitis (CARE), one of gastroesophageal reflux disease (GERD) is increasing worldwide. Coptidis rhizoma extract (CRE) is a traditional herb that cures a variety of diseases. This study was conducted to evaluate the protective effect of CR on rats with chronic acid reflux esophagitis. Methods : The antioxidant activities were evaluated through radical scavenging assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) radical scavenging assays. CARE was surgically induced in 5-week-old male SD rats by ligating the border between forestomach and glandular portion with a 2-0 silk tie and covering the duodenum using 18-Fr $N{\acute{e}}laton$ catheter. To evaluate the esophageal protective effect of CRE, rats were divided into 3 groups: Nor (normal rats), Veh (chronic acid reflux esophagitis induced rats), CR (chronic acid reflux esophagitis induced rats treated with CRE 200 mg/kg body weight). Results : The administration of CRE significantly prevented the mucosal injury of the esophagus tissue and histological findings improved the esophageal lesion. It has been shown that inflammation is prevented by the increase of antioxidant-related factors (Nrf-2, HO-1, SOD, catalase, and GPx-1/2) through the antioxidant pathway of esophageal tissue. The administration of CRE reduced the increase of serum peroxynitrite ($ONOO^-$) and markedly reduced the protein expression of inflammatory mediator such as $NF-{\kappa}Bp65$, $p-I{\kappa}B{\alpha}$, iNOS, and IL-6. Conclusions : Overall, these results suggest that CRE administration confirmed the protective effect of esophageal mucosa, suggesting that it is a potential treatment for chronic acid reflux esophagitis.

• 대한의생명과학회지
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• 제18권2호
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• pp.104-111
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• 2012

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases which degrade extracellular matrix (ECM) during embryogenesis, wound healing, and tissue remodeling. Dysregulation of MMP activity is also associated with various pathological inflammatory conditions. In this study, we examined the expression pattern of MMPs during PMA-induced differentiation of THP-1 monocytic cells into macrophages. We found that MMP1, MMP8, MMP3, MMP10, MMP12, MMP19, MMP9, and MMP7 were upregulated during differentiation whereas MMP2 remained unchanged. Expression of MMPs increased in a time-dependent manner; MMP1, MMP8, MMP3, MMP10, and MMP12 increased beginning at 60 hr post PMA treatment whereas MMP19, MMP9, and MMP7 increased beginning at 24 hr post PMA treatment. To identify signal transduction pathways involved in PMA-induced upregulation of MMPs, we treated PMA-differentiated THP-1 cells with specific inhibitors for PKC, MEK1, NF-${\kappa}B$, PI3K, p38 MAPK and PLC. We found that inhibition of the MEK1 pathway blocked PMA-induced upregulation of all MMPs to varying degrees except for MMP-2. In addition, expression of select MMPs was inhibited by PI3K, p38 MAPK and PLC inhibitors. In conclusion, we show that of the MMPs examined, most MMPs were up-regulated during differentiation of monocyte into macrophage via the MEK1 pathway. These results provide basic information for studying MMPs expression during macrophage differentiation.

• Asian-Australasian Journal of Animal Sciences
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• 제30권10호
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• pp.1471-1477
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• 2017

Objective: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusion: It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an $NF-{\kappa}B$ signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.

• Tuberculosis and Respiratory Diseases
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• 제57권5호
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• pp.449-460
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• 2004

연구배경 : PS-341은 최근에 개발된 강력하고 특이적인 proteasome 억제제로서, 일부 암환자에 투여하여 좋은 성적이 보고되고 있다. Proteasome 억제제의 항암효과는 아포프토시스 유발 물질, 즉 p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax, Bcl-2 등의 발현 증가와 관련이 있는 것으로 생각되고 있다. JNK와 GSK-$3{\beta}$도 아포프토시스에 관여하는 것으로 잘 알려져 있지만, PS-341에 의한 아포프토시스에서의 역할은 규명되지 못한 상태이다. 본 연구에서는 폐암세포주에서 PS-341에 의한 아포프토시스에서 JNK와 GSK-$3{\beta}$의 역할을 규명하고자 하였다. 방 법 : NCI-H157과 A549 폐암세포주를 실험에 사용하였다. 세포생존능은 MTT 방법으로 평가하였고, 아포프토시스는 PARP의 분해로 평가하였다. JNK의 활성도는 in vitro immuno complex kinase 방법과 내인성 c-Jun의 인산화로 측정하였다. 각종 단백의 발현은 Western 분석으로 평가하였다. JNK1과 GSK-$3{\beta}$의 과발현은 각각 plasmid vector와 adenovirus vector를 이용하였다. 결 과 : PS-341 처치로 아포프토시스에 의한 세포생존율의 감소가 관찰되었다. PS-341 처치로 JNK가 활성화되었고, c-Jun의 발현이 유도되었다. Dominant negative JNK1의 과발현 또는 SP600125 전치치로 JNK의 활성화를 차단하면 PS-341에 의한 아포프토시스가 억제되었다. PS-341 처리로 JNK 활성화에 의존적으로 caspase 3의 활성화가 유도되었다. Caspase 활성화의 차단으로도 PS-341에 의한 아포프토시스가 억제되었다. PS-341에 의해 Akt가 활성화되었고, Akt 활성화의 차단으로 PS-341에 의한 아포프토시스가 심화되었다. PS-341에 의한 Akt 활성화로 GSK-$3{\beta}$가 불활성화되었다. Constitutively active GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 심화되었고, dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 감소되었다. Lithium chloride 전처치와 dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 JNK의 활성화와 c-Jun의 발현 증가가 억제되었다. 결 론 : 폐암세포주에서 PS-341에 의한 아포프토시스는 JNK/caspase 경로가 관여하며, 이는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화에 의해 억제되는 것으로 판단된다. 따라서 PS-341의 항암효과를 최대화하기 위해서는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화를 차단하는 치료법이 병행되어야 할 것으로 판단된다.

• Gut and Liver
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• 제12권6호
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• pp.682-693
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• 2018

Background/Aims: Intestinal barrier dysfunction is a hallmark of inflammatory bowel diseases (IBDs) such as ulcerative colitis. This dysfunction is caused by increased permeability and the loss of tight junctions in intestinal epithelial cells. The aim of this study was to investigate whether estradiol treatment reduces colonic permeability, tight junction disruption, and inflammation in an azoxymethane (AOM)/dextran sodium sulfate (DSS) colon cancer mouse model. Methods: The effects of $17{\beta}$-estradiol (E2) were evaluated in ICR male mice 4 weeks after AOM/DSS treatment. Histological damage was scored by hematoxylin and eosin staining and the levels of the colonic mucosal cytokine myeloperoxidase (MPO) were assessed by enzyme-linked immunosorbent assay (ELISA). To evaluate the effects of E2 on intestinal permeability, tight junctions, and inflammation, we performed quantitative real-time polymerase chain reaction and Western blot analysis. Furthermore, the expression levels of mucin 2 (MUC2) and mucin 4 (MUC4) were measured as target genes for intestinal permeability, whereas zonula occludens 1 (ZO-1), occludin (OCLN), and claudin 4 (CLDN4) served as target genes for the tight junctions. Results: The colitis-mediated induced damage score and MPO activity were reduced by E2 treatment (p<0.05). In addition, the mRNA expression levels of intestinal barrier-related molecules (i.e., MUC2, ZO-1, OCLN, and CLDN4) were decreased by AOM/DSS-treatment; furthermore, this inhibition was rescued by E2 supplementation. The mRNA and protein expression of inflammation-related genes (i.e., KLF4, NF-${\kappa}B$, iNOS, and COX-2) was increased by AOM/DSS-treatment and ameliorated by E2. Conclusions: E2 acts through the estrogen receptor ${\beta}$ signaling pathway to elicit anti-inflammatory effects on intestinal barrier by inducing the expression of MUC2 and tight junction molecules and inhibiting pro-inflammatory cytokines.