• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.1947-1959
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• 2016

Background: Colorectal cancer (CRC) is a major cause of morbidity and mortality, being the second most common type of cancer worldwide in both men and women. It accounts yearly for approximately 9% of all new cases of cancers. Furthermore, the current chemotherapeutic regimens seem unsatisfactory, so that exploration of novel therapeutic modalities is needed. The present study was undertaken to investigate the inhibitory effects of a crude alkaloid extract (CAERS) of a medicinal herb, Rhazya stricta, on proliferation of CRC HCT116 cells and to elucidate mechanisms of action. To achieve these aims, we utilized MTT, comet, DNA laddering and gene reporter assays, along with Western blot and RT-PCR analyses. Results: We found that CAERS inhibited cell proliferation and induced apoptotic cell death in HCT116 cells. Hallmarks of morphological and biochemical signs of apoptosis were clearly evident. CAERS down-regulated DNA-binding and transcriptional activities of NF-${\kappa}B$ and AP-1 proteins, while up-regulating expression of the Nrf-2 protein. It also down-regulated expression levels of the ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin and VEGF and up-regulated levels of Bax, caspase-3/7 and -9, p53, p21, Nrf-2. Markedly, it promoted mRNA expression levels of cytoprotective genes including the hemeoxygenase-1, NAD(P)H quinine oxidoreductase 1 and UDP-glucuronyltransferase. Conclusions: These findings indicate that CAERS exerts antiproliferative action on CRC cells through induction of apoptotic mechanisms, and suggest CAERS could be a promising agent for studying and developing novel chemotherapeutic agents aimed at novel molecular targets for the treatment of CRC.

• 생약학회지
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• 제42권2호
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• pp.175-181
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• 2011

Oxidative stress or accumulation of reactive oxygen species (ROS) leads neuronal cellular death and dysfunction, and it contributes to neuronal degenerative disease such as Alzheimer's disease, Parkinson's disease and stroke. Glutamate is one of the major excitatory neurotransmitter in the central nervous system (CNS). Glutamate contributes to fast synaptic transmission, neuronal plasticity, outgrowth and survival, behavior, learning and memory. In spite of these physiological functions, high concentration of glutamate causes neuronal cell damage, acute insults and chronic neuronal neurodegenerative diseases. Heme oxygenase-1 (HO-1) enzyme plays an important role of cellular antioxidant system against oxidant injury. NNMBS020, the water-insoluble fraction of the 70% EtOH extract of root barks of Dictamnus dasycarpus, showed dominant neuroprotective effects on glutamate-induced neurotoxicity in mouse hippocampal HT22 cells by induced the expression of HO-1 and increased HO activity. In mouse hippocampal HT22 cells, NNMBS020 makes the nuclear accumulation of Nrf2 and stimulates extracellular signal-regulated kinase (ERK) pathway. The ERK MAPK pathway inhibitor significantly reduced NNMBS020-induced HO-1 expression, whereas the JNK and p38 inhibitors did not. In conclusion, the water-insoluble fraction of the 70% EtOH extract of root barks of D. dasycarpus (NNMBS020) significantly protect glutamate-induced oxidative damage by induction of HO-1 via Nrf2 and ERK pathway in mouse hippocampal HT22 cells.

• 생명과학회지
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• 제28권2호
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• pp.207-215
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• 2018

염증 반응과 산화적 스트레스는 다양한 질환의 발생과 진행에 중요한 역할을 담당한다. 따라서 염증과 산화적 스트레스를 동시에 억제할 수 있는 소재의 발굴은 인체 질환 제어에 매우 유용하게 적용될 수 있다. 본 연구의 목적은 RAW 264.7 대식세포에서 검은콩 품종인 소청자와 소청2호 추출물이 염증성 및 산화적 스트레스에 미치는 영향을 알아보기 위함이다. 본 연구의 결과에 의하면 소청자 및 소청2호 추출물은 LPS에 의한 iNOS와 COX-2의 발현을 억제하여 NO와 $PGE_2$의 생성을 억제하였으며, $TNF-{\alpha}$, $IL-1{\beta}$와 같은 염증성 cytokine의 분비와 발현을 감소시켰다. 또한 소청자 및 소청2호 추출물은 LPS로 자극된 세포 내 ROS 축적을 유의적으로 차단시켰고, Nrf2와 HO-1 발현을 증가시켰다. 또한 LPS에 의해 유도된 MAPKs의 활성화도 소청자 및 소청2호 추출물에 의하여 억제되었다. 결론적으로 소청자 및 소청2호 추출물은 RAW 264.7 대식세포에서 MAPKs 경로를 차단시킴으로써 LPS-유도 염증 및 산화 반응에 대한 보호 역할을 할 수 있으며, 이러한 효과에는 최소한 Nrf2/HO-1 경로 활성화가 관련되어 있었다. 이러한 결과를 고려해 볼 때 소청자 및 소청2호 추출물은 대식세포의 과다 활성화로 인한 염증성 및 산화적 반응 차단에 잠재적인 효과가 있음을 일 수 있었다.

• 대한한의학방제학회지
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• 제31권1호
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• pp.29-39
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• 2023

Objective : Gardeniae Fructus (GF) is the ripe fruit of Gardenia jasminoides Ellisa with a bitter taste and cold properties. Ingredient compounds including geniposide are known to have anti-inflammatory, antioxidant, and neuroprotective effects. The purpose of this study was to investigate the neuroprotective effect of GF on tBHP-induced PC12 cells. Methods : Cell viability was measured by the MTT assay, and apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The expression level of each protein was monitored by Western blot analysis, and reactive oxygen species (ROS) were analyzed using DCFH-DA. Results : In PC12 cells, tBHP induced cell death through apoptosis with caspase activation and PARP inactivation. Cells treated with tBHP showed an increase in intracellular ROS and depletion of GSH. Pretreatment with GF prevented tBHP-induced apoptosis, reduced ROS, and increased GSH. GF also maintained increased Nrf2 expression in the presence of tBHP. Phosphorylation of JNK and p38 MAPK was increased by tBHP, whereas phosphorylation of ERK was decreased. GF restored changes in ERK and p38 phosphorylation, but not JNK phosphorylation. Conclusion : These results indicate that GF has neuroprotective effects through anti-apoptotic and antioxidant effects mediated by regulation of Nrf2 expression and phosphorylation of ERK and p38. It also demonstrates the potential use of GF as a source of antioxidant and neuroprotective substances.

• 생약학회지
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• 제51권4호
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• pp.349-359
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• 2020

Gastroesophageal reflux disease (GERD) is a disease that stomach contents continually refluxing, and is currently on the rise worldwide. The purpose of this study is to find natural materials that can reduce side effects and effectively treat chronic acid reflux esophagitis (CARE), one of GERD. First, the antioxidant activity was confirmed by varying the mixing ratio of Coptidis Rhizoma and Evodiae Fructus, which are effective against chronic reflux esophagitis. After, animal experiments were conducted using a 1:1 (CE) and 1:2 (CEE) combination ratio of Coptidis Rhizoma and Evodiae Fructus, which had the best antioxidant efficacy. Gross lesion of esophageal mucosa after CE or CEE treatment showed a superior enhancement compared with that of CARE control rats. Additionally, its inhibited MAPK phosphorylation and led NF-κB inactivation through the suppression of IκBα phosporylation by regulating Nrf2/Keap-1, and NF-κB inactivation induced reduced protein expressions including inflammatory mediators and cytokines. Moreover, its improved esophageal barrier function through upregulating protein expressions of tight junction protein, whereas downregulating protein expressions of MMPs. Taken together, a mixture of Coptidis Rhizoma and Evodiae Fructus can attenuate the esophageal mucosal ulcer by inhibiting MAPK and NF-κB pathway, and upregulating proteins associated with tight junction.

• 생명과학회지
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• 제27권12호
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• pp.1437-1444
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• 2017

본 연구에서는 갈조류인 큰열매모자반 에탄올 추출물(Ethanol extract of Sargassum Macrocarpum, EESM)의 항염증 활성을 조사하였다. Lipopolysaccharide (LPS)로 염증이 유도된 RAW 264.7 대식세포에서 EESM에 의한 항염증 효과를 조사한 결과 EESM은 nitric oxide (NO)와 prostaglandin $E_2$ ($PGE_2$)의 분비를 억제하는 결과를 보였고, 이는 inducible NO synthase (iNOS)와 cyclooxygenase-2 (COX-2)의 발현 역시 억제하였다. 또한 전염증성 사이토카인인 tumor necrosis factor-alpha ($TNF-{\alpha}$)와 interleukin 1 beta ($IL-1{\beta}$)의 발현도 억제하였다. 이러한 염증반응이 어떤 신호경로를 통해 일어나는지 알아보기 위해 nuclear factor kappa B ($NF-{\kappa}B$), phosphoinositide-3-kinase (PI3K)/Akt 그리고 mitogen-activated protein kinases (MAPKs) 신호전달 경로를 조사한 결과 EESM에 의해 $NF-{\kappa}B$가 세포질에서 핵으로 이동과 Akt 단백질의 인산화가 억제되었고 MAPK에 속하는 kinase 중 ERK의 단백질 인산화가 억제되었다. 따라서 EESM이 $NF-{\kappa}B$ 신호계를 포함한 PI3K/Akt 및 MAPK/ERK 신호경로를 동시에 관여하고 있음을 알 수 있었다. 또한 heme oxygenase-1 (HO-1)과 그 전사 인자인 nuclear factor-E2-related factor 2 (Nrf2)의 단백질 발현에 미치는 영향을 분석한 결과 EESM에 의해 HO-1 및 Nrf2의 발현이 증가됨을 보였다. 본 연구 결과 EESM은 높은 항염증 활성을 갖는 것으로 확인되였으며 향후 잠재적인 기능성 소재로서 유용하게 활용될 수 있을 것으로 사료된다.

• 생명과학회지
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• 제29권3호
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• pp.325-331
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• 2019

상추는 가장 선호하는 녹색 채소 중 하나이다. 상추는 폴리페놀성 화합물을 비롯한 다양한 성분을 함유하고 있으며, 항균, 항산화, 항염증 등의 생리활성을 가지고 있는 것으로 알려져 있다. 본 연구에서는 건조상추의 에탄올 추출물(DLE)을 제조하고 이들의 항염증 활성을 연구하였다. DLE의 항염증 활성을 측정하기 위하여 LPS로 활성화된 마우스 대식세포 RAW 264.7 세포주에서 nitric oxide (NO) 생성을 측정하였다. DLE는 세포주의 생존에는 영향을 미치지 않으면서 NO 생산을 현저하게 저해하였다. DLE에 의해 염증 유전자인 iNOS와 COX-2의 유전자와 단백질의 발현이 모두 감소하였으며, 6개의 염증관련 cytokine 유전자($IL-1{\alpha}$, $IL-1{\beta}$, IL-1F6, $TNF-{\alpha}$, CSF2, 그리고 CXCL10)의 발현이 모두 감소하였다. 또한, DLE의 처리는 MAPKs 경로의 인산화를 모두 저해하였으며, $NF-{\kappa}B$ p65의 핵으로의 이동을 저해하였다. 이러한 결과는 DLE의 항염증 활성은 MAPKs 경로와 $NF-{\kappa}B$ 경로를 조절함으로써 이루어짐을 시사한다. 또한, DLE는 농도의존적으로 reactive oxygen species (ROS)의 생산을 저해하였으며, hemeoxygenase-1 (HO-1) 단백질의 발현을 증가시켰으며, HO-1의 전사조절인자인 Nrf2의 핵으로의 이동을 증가시켰다. 결론적으로, 이러한 연구결과는 DLE가 염증관련 유전자의 발현을 감소시키며, MAPKs, $NF-{\kappa}B$, 그리고 Nrf2/HO-1 등 다양한 경로를 조절함으로써 항염증 활성을 가지는 것을 제시한다.

• 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.

• Journal of Nutrition and Health
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• 제44권6호
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• pp.488-497
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• 2011

Oxygen is necessary to sustain life, yet cellular oxygen metabolism creates destructive elements called free radicals. Free radicals are chemically unbalanced and carrying free electrons that can damage molecules, potentially damaging the cell itself. For this reason, many antioxidant products, including supplements and functional foods, are being developed. In particular, natural products are rich sources of pharmacologically active compounds. The purpose of this study was to investigate the antioxidant effects of target biomaterials in Korean traditional spices such as diallyl sulfide (DAS), capsaicin (CAP), and gingerol (GGR), and to investigate the response of the antioxidant defense system to oxidative stress by hydrogen peroxide ($H_2O_2$) compared to sulforaphane (SFN) in HepG2 cells. After the analysis of the cell viability using Cell Counting kit-8 (CCK-8) assay, we determined that the optimum levels were $200{\mu}M$ DAS, $25{\mu}M$ CAP, $50{\mu}M$ GGR, and $12.5{\mu}M$ SFN. Antioxidant enzymes were measured and protein expression was detected by Western blotting. All treatments showed a significant decrease in antioxidant enzyme activity such as superoxide dismutase, catalse, and glutathione peroxidase in HepG2 cells. Additionally, DAS, CAP, GGR and SFN increased the antioxidant system-related transcription factor Nrf2 which was found to be regulated by the activation of MAPK-JNK in this study. In conclusion, these results indicate the protective effects of DAS CAP, GGR, and SFN against $H_2O_2$-induced oxidative stress.

• The Korean Journal of Physiology and Pharmacology
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• 제20권3호
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• pp.229-236
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• 2016

Resveratrol (RSV) may provide numerous protective effects against chronic inflammatory diseases. Due to local hypoxia and hypertonicity, the renal medulla is subject to extreme oxidative stress, and aldehyde products formed during lipid peroxidation, such as 4-hydroxy-2-hexenal (HHE), might be responsible for tubular injury. This study aimed at investigating the effects of RSV on renal and its signaling mechanisms. While HHE treatment resulted in decreased expression of Sirt1, AQP2, and nuclear factor erythroid 2-related factor 2 (Nrf2), mouse cortical collecting duct cells (M1) cells treated with HHE exhibited increased activation of p38 MAPK, extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and increased expression of NOX4, $p47^{phox}$, Kelch ECH associating protein 1 (Keap1) and COX2. HHE treatment also induced $NF-{\kappa}B$ activation by promoting $I{\kappa}B-{\alpha}$ degradation. Meanwhile, the observed increases in nuclear $NF-{\kappa}B$, NOX4, $p47^{phox}$, and COX2 expression were attenuated by treatment with Bay 117082, N-acetyl-l-cysteine (NAC), or RSV. Our findings indicate that RSV inhibits the expression of inflammatory proteins and the production of reactive oxygen species in M1 cells by inhibiting $NF-{\kappa}B$ activation.