• 한국미생물·생명공학회지
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• 제42권3호
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• pp.275-284
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• 2014

본 연구에서는 식물자원으로부터 생리활성을 보유한 새로운 기능성 소재를 찾고자 하였다. 이를 위해 수종의 중국 자생 식물을 분석하여 강한 항산화능을 보유한 3종(Malus hupehensis, Ophiorrhiza cantonensis, 그리고 Psychotria rubra)을 선별하고, 각 추출물의 항산화능과 항염증 생리활성을 분석하였다. 먼저 각 추출물의 항산화능을 DPPH radical 소거능을 통해 분석한 결과 모두 양성 대조군으로 사용한 ascorbic acid와 유사한 정도의 높은 활성을 보여 각 소재가 매우 강한 항산화능을 보유함을 확인하였다. 또한 RAW 264.7 세포주에서 $H_2O_2$ 및 LPS에 의해 유도된 ROS에 대한 각 추출물의 소거능을 분석한 결과에서도 농도의존적인 강한 소거능을 보였다. 뿐만 아니라 대표적인 항산화효소 중 하나로 항산화능 보유 천연물에 의해 발현이 유도되는 HO-1 및 그 전사 인자인 Nrf-2의 단백질 발현이 각 추출물의 처리에 의해 증가됨을 보였다. 한편 각 소재가 LPS에 의해 유도된 NO 생성에 미치는 영향을 분석한 결과 모두 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 확인하였다. 이와 같은 각 소재의 NO 생성 및 iNOS 발현 억제 효과는 염증 상위신호 전달계인 NF-${\kappa}B$ 및 AP-1의 조절을 통해 일어날 가능성을 보였다. 이러한 결과를 통해 중국 자생 식물 3종의 높은 항산화능과 항염증 활성을 처음으로 확인하였으며 향후 기능성 소재로서 유용하게 활용될 수 있을 것으로 판단된다.

• 한방재활의학과학회지
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• 제33권2호
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• pp.1-18
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• 2023

Objectives The purpose of this study was to evaluate the antioxidant, anti-inflammatory and wound healing effects of Samul-tang-ga-dansam water extract (SD) in wound-induced mice. Methods The mice were divided into five groups (n=7): the normal group, the control group, the positive control group, the low-dose SD group and the high-dose SD group. The normal group had no wounds and the other groups were wounded on the back with a leather punch. Distilled water was administered to the control group, 200 mg/kg of vitamin E was administered to the positive control group. In the low-dose SD group and the high-dose SD group, 1.23 g/kg and 2.47 g/kg of SD were administered, respectively. Antioxidant and anti-inflammatory protein levels were evaluated using western blot analysis. Skin tissue was analyzed by H&E, Masson's trichrome staining method. Results Oral administration of the SD significantly reduced the visible skin damage and decreased the reactive oxygen species and ONOO^- activity of the serum. It significantly increased heme oxygenase-1, superoxide dismutase, catalase, GPx-1/2, Nrf2 and Keap-1 which are antioxidant-related factors in skin tissue and reduced NF-κB p65, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor α, interleukin (IL)-1β, IL-6 which are inflammation-related factors. Also, SD significantly decreased NOX2, p22^phox and p47^phox and increased α-smooth muscle actin and COL1A1 protein expression in fibroblasts involved in connective tissue repair. According to histological examination, the thickened epithelial layer was thinned and collagen fibers were increased to accelerate wound healing. Conclusions It is suggested that Samul-tang-ga-dansam has antioxidant and anti-inflammatory effects and promotes wound tissue repair.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5231-5235
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• 2013

Objective: NF-E2-related factor 2 (Nrf2) is activated in several human malignancies. However, the role of Nrf2 in gastric cancer (GC) remains incompletely understood. In this study, we therefore analyzed associations of Nrf2 expression status with clinical features and chemotherapeutic resistance in GC. Materials and Methods: A total of 186 samples from GC patients who underwent gastrectomy were used for prognostic assessment. A further 142 samples from GC cases who received first-line combination chemotherapy were applied for investigation of chemoresistance. The Nrf2 expression was evaluated by immunohistochemistry in GC samples, and its relationship with clinicopathological parameters and chemotherapy sensitivity was analyzed. The effect of Nrf2 gene silencing on chemotherapy resistance was also examined by cell viability assay in vivo. Results: Of the 186 patients with GC, 104/186 (55.9%) showed high expression for Nrf2. The overexpression of Nrf2 was an independent predictor of overall survival [OS, hazard ratio (HR) 3.9; P=0.011] and disease-free survival (DFS, HR 4.3; P=0.002). The gene silencing of Nrf2 reduced resistance to cell death induced by 5-FU in GC cell lines. Conclusion: Our data show that Nrf2 is an independent prognostic factor in GC. Furthermore, Nrf2 confers resistance to chemotherapeutic drug 5-FU in GC cells. Taken together, Nrf2 is a potential prognostic marker and predictive for 5-FU resistance in GC.

• 농업생명과학연구
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• 제50권2호
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• pp.139-150
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• 2016

말뼈추출물은 다양한 골질환의 예방과 치료에 탁월한 효능이 있다고 이전에 보고되었다. 하지만 말뼈추출물의 다른 약리학적 효능에 대해서는 아직 자세히 밝혀지지 않고있다. 본 연구에서는 말뼈추출물이 중요한 항산화 인자인 hemeoxygenase-1(HO-1)의 발현을 상승시킬 수 있는지, 만약 발현이 증가한다면 HO-1의 상향 조절이 대식세포에서 항염증 효과를 매개할 수 있는지에 관하여 조사하였다. 이를 위해서 nitric oxide(NO) 농도측정, 세포 생존능 측정, DPPH 라디칼 소거능 검사를 시행하였다. 또한 염증성 사이토카인 유전자 발현과 단백질 발현을 측정하기 위해 real time PCR과 Western blotting을 시행하였다. 말뼈추출물은 lipopolysaccharide(LPS, 0.1㎍/ml)로 자극한 대식세포주인 RAW264.7 세포에서 어떠한 세포독성 없이 NO의 생성을 유의성 있게 억제하였으며 inducible nitric oxide(iNOS)와 cyclooxygenase 2(COX-2)의 발현을 억제하였다. 뿐만 아니라 말뼈 추출물은 염증성 사이토카인인 tumor necrosis factor(TNF)-α와 interleukin(IL)-1β의 발현을 억제하였으며 ERK, JNK 및 p-38 MAPK의 단백질 인산화를 억제하였다. 그리고 말뼈추출물은 HO-1과 NF-E2-related factor-2(Nrf-2) 의 발현을 증가시켰고 이것은 말뼈추출물이 가지고 있는 항 염증효과를 매개할 수 있는 것으로 보인다. 즉, 말뼈추출물이 HO-1의 발현을 상향 조절한 반면 ERK1/2의 신호전달 경로에 손상을 주는 것으로 확인되었으며 이러한 말뼈추출물의 효과가 최종적으로 세포손상과 세포의 과산화 자극으로부터 세포를 보호 할 수 있는 것으로 사료된다.

• 대한본초학회지
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• 제30권6호
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• pp.7-15
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• 2015

Objectives : Rosa laevigata Michx. has been used for the treatment of renal disease in traditional Korean medicine. In this study, we investigated cytoprotective effect of R. laevigata water extract (RLE) against oxidative stress induced by arachidonic acid (AA) + iron.Methods : To evaluate the protective effects of RLE against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The effects of RLE on reduced glutathione level, production of reactive oxygen species and mitochondrial membrane potential were also monitored. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) was examined by immunoblot analysis. Additionally, Nrf2 transactivation and its downstream target genes expression were also determined by reporter gene and realtime RT-PCR analyses.Results : RLE pretreatment (30-300 μg/ml) prevented cells from AA + iron-mediated cell death in a concentration dependent manner. In addition, 100 μg/ml RLE inhibited AA + iron-induced glutathione depletion, reactive oxygen species production and mitochondrial dysfunction. RLE accumulated nuclear Nrf2 and also transactivated Nrf2, which was evidenced by antioxidant response element- and glutathione S-transferase A2-driven luciferase activities and mRNA level of glutamate-cysteine ligase catalytic subunit, NAD(P)H:quinone oxidoreductase 1 and sestrin 2. Moreover, protective effect of RLE against AA + iron was abolished in Nrf2 knockout cells.Conclusions : These results indicate that RLE has the ability to protect hepatocyte against oxidative stress through Nrf2 activation.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1331-1338
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• 2010

This study was designed to investigate the potentially protective effects of Curcuma longa Linn. extract (CLE) on carbon tetrachloride ($CCl_4$)-induced hepatotoxicity in rats. Male Sprague-Dawley rats were pretreated with 50 or 100mg/kg of CLE or 100mg/kg of butylated hydroxytoluene(BHT) for 14 days before $CCl_4$ administration. In addition, the CLE control group was pretreated with 100mg/kg CLE for only 14 days. Three hours after the final treatment, a single dose of $CCl_4$ (20mg/kg) was administrated intraperitoneally to each group. After the completion of this phase of the experiment, food and water were removed 12 h prior to the next step. The rats were then anesthetized by urethane and their blood and liver were collected. It was observed that the aspartate aminotransferase and alanine aminotransferase activities of the serum, and the hepatic malondialdehyde levels had significantly decreased in the CLE group when compared with the $CCl_4$-treated group. The antioxidant activities, such as superoxide dismutase, catalase, and glutathione peroxidase activities, in addition to glutathione content, had increased considerably in the CLE group compared with the $CCl_4$-treated group. Phase II detoxifying enzymes, such as glutathione S-transferase, were found to have significantly increased in the CLE group as opposed to the $CCl_4$-treated group. The content of Nrf2 was determined by Western blot analysis. Pretreated CLE increased the level of nuclear translocated Nrf2, and the Nrf2 then increased the activity of the antioxidant and phase II detoxifying enzymes. These results indicate that CLE has protective effects against $CCl_4$-induced hepatotoxicity in rats, via activities of antioxidant and phase II detoxifying enzymes, and through the activation of nuclear translocated Nrf2.

• 대한본초학회지
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• 제30권6호
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• pp.63-68
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• 2015

Objectives : Gastric lesions affect many people around the world and their development are results of the imbalance between destructive and protective factors in the gastric mucosa. Lycium chinense has been widely used as a traditional Korean medicine, it was recently reported that they have potent anti-inflammatory effects in chronic hepatitis models. Therefore, this study aimed to investigate the anti-inflammatory activity of Lycium chinense extract (LCE) on HCl-Ethanol induced gastric lesion mice.Methods : The ICR mice were divided randomly into five groups of six animals each. Group A was normal mice, and group B was treated orally with 0.5 ml 150 mM HCl-60% Ethanol. Mice in group C and D were pre-treatment of LCE (100 mg/kg and 200 mg/kg bodyweight, p.o before HCl/ethanol treatment) and group E was orally administered sucralfate (10 mg/kg).Results : 150mM HCl/60% ethanol-induced gastric mucosal injury mice were ameliorated mucosal damage upon histological evaluation by treatment of LCE. Pre-treatment of LCE attenuated reactive oxidative species (ROS) and produces peroxynitrite (ONOO^-) in stomach tissues. As results of stomach protein analyses, LCE effectively reduce inflammatory-related factors such as cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), inducible nitric oxide synthase (iNOS) and interleukin-6 (IL-6) in gastric lesion mice. In addition, nuclear factor kappa B (NF-κB) and inhibitor of phosphorylation of nuclear factor kappa B (p-IκB) were down-regulated in LCE-administrated gastric lesion mice.Conclusions : Our discovery supports that the therapeutic activity of LCE ameliorate the development of gastric lesion via suppressing the oxidative stress and gastric partial inflammation induced by 150 mM HCl/60% ethanol.

• Molecular & Cellular Toxicology
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• 제5권2호
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• pp.120-125
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• 2009

Alaria esculenta is a brown seaweed found in the Arctic. This study investigated the protective effect of A. esculenta extract (AEE) against oxidant-mediated injury and its mode of action in RAW264.7 macrophages. The methyl thiazolyl tetrazolium (MTT) assay showed that $H_2O_2$ treatment reduced cell viability, whereas AEE protected cells from $H_2O_2$-mediated cytotoxicity in a dose-dependent manner. Because heme oxygenase-1 (HO-1) is known to protect cells against oxidative damage, we investigated the effect of AEE on HO-1 gene expression and HO enzyme activity. The protective effect of AEE against $H_2O_2$-induced injury was correlated with increased HO enzyme activity. AEE also induced HO-1 mRNA and protein expression, as determined RT-PCR and Western blotting, respectively. To characterize the mechanisms by which AEE induces HO-1 gene expression, we examined the effect of AEE on the nuclear translocation of NF-E2-related factor-2 (Nrf2) and Akt phosphorylation. AEE treatment activated upstream signaling for HO-1 gene expression, including the nuclear translocation of Nrf2 and Akt phosphorylation. Collectively, these results suggest that AEE has anti-oxidant activity that is mediated, at least in part, via the activation of Nrf2 and Akt and the subsequent induction of HO-1 gene expression.

• 한방안이비인후피부과학회지
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• 제26권2호
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• pp.45-57
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• 2013

Objectives : Daucus carota sativa has been frequently used as food supplements in many of the Asian countries, and a nutritional medical drug in traditional medicine. This research investigated the effects of Daucus carota sativa extract (DCE) on acute phases of inflammation in Raw 264.7 cells treated with lipopolysaccharide (LPS) in terms of the inhibition of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines production. Methods : NO, $PGE_2$, tumor necrosis factor (TNF)-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6 contents were assayed by ELISA, and expressions of inflammation-related proteins such as inducible NO synthase (iNOS) were determined by immunoblot analyses. Results : DCE treatment attenuated the LPS ability to increase the productions of NO and $PGE_2$ as well as the protein level of iNOS in a concentration-dependent manner. Consistently, treatment of the cells with DCE suppressed the production of TNF-${\alpha}$, interleukin-$1{\beta}$ and interleukin-6. DCE also caused decreases of inhibitor of ${\kappa}B{\alpha}$ phosphorylation induced by LPS in the cells, which means DCE inhibition of NF-${\kappa}B$ activity. Furthermore, DCE blocked LPS-induced phosphorylation of p38 and SAPK/JNK. Conclusion : This study showing here may be of help to understand the action mechanism of DCE, and provide the information for the medical use of Daucus carota sativa for the inflammatory disease.

• 한국식품저장유통학회지
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• 제25권1호
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• pp.107-116
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• 2018

본 연구의 목적은 블루베리 잎의 용매별 추출물에 따른 약리활성에 대한 검증 및 효능 평가이다. 블루베리 잎의 항산화, 항염증에 대한 효과를 확인하였다. 염증 반응은 자극이 가해지면 histamine, serotonin, prostaglandin과 같은 혈관 활성물질에 의해 혈관 투과성이 증대되어 염증을 유발하고 cytokine, free radical, lysosomal enzyme 등 다양한 매개 인자가 관여한다. 자극에 의한 macrophage cell의 염증반응은 tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin-6(IL-6), interleukin-$1{\beta}$(IL-$1{\beta}$)와 같은 pro-inflammatory cytokine의 발현이 유도되고, inducible nitric oxide synthase(i-NOS)와 cyclooxygenase-2(COX-2)에 영향을 받는 유전자의 발현을 자극하게 되어 nitric oxide(NO) 및 $PGE_2$등의 염증 인자가 생성된다. 이에 따라 블루베리 잎 추출물의 항염증에 대한 연구를 위해 이에 영향을 주는 인자인 i-NOS, COX-2의 단백질 발현억제 작용을 확인 하였다. 그 결과 BLA > BLE > BLW 순서로 높은 효능을 확인 할 수 있었다. 가장 효과가 좋은 BLA 처리군에서 $PGE_2$ 분비량 및 다양한 염증성 인자의 mRNA 발현량을 확인하였다. 측정 결과, BLA($100{\mu}g/mL$)는 $PGE_2$ 분비량을 85.3% 억제하였으며 i-NOS, COX-2, IL-$1{\beta}$, IL-6, TNF-${\alpha}$ 단백질과 mRNA 발현이 각각 86.8%, 85.7%, 62.7%, 77%, 32.2% 억제되는 효과를 확인할 수 있었다. 결론적으로 블루베리 70% 아세톤 추출물(BLA)의 항염증 효과가 가장 높았으며, 블루베리 잎은 세포의 mRNA 및 단백질 수준에서 염증인자들의 억제를 통해 대식세포에서 항염증 효과가 명백히 확인되었다. 더 나아가, nuclear factor kappa B(NF-${\kappa}B$), signal transducer and activator of transcription(STAT-1), mitogen-activated protein kinases(MAPKs) 등의 세포 내 염증관련 중간기전을 연구해볼 가치가 있다고 사료된다.