• 생명과학회지
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• 제25권9호
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• pp.976-983
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• 2015

Achyranthoside C dimethyl ester (ACDE)는 우슬에서 분리한 oleanolic acid glycoside이다. 본 연구에서는 RAW264.7 대식세포에서 ACDE의 항염증 효과를 관찰하고 그 작용 기전을 연구하였다. ACDE는 세포에 독성을 유도하지 않으면서 heme oxygenase-1 (HO-1)의 발현을 유도하였다. ACDE 는 HO-1의 발현에 관여하는 전사인자인 nuclear factor E2-related factor 2 (Nrf2)를 핵으로 이동시켰다. 또한 ACDE에 의한 HO-1의 발현은 phosphatidylinositol 3-kinase (PI-3K) 및 mitogen activated protein kinases (MAPK) 억제제에 의해 감소되었으며, ACDE가 Akt, c-Jun kinase (JNK), extracellular signal regulated kinase (ERK), p38 kinase의 인산화를 유도하였다. 한편 ACDE는 lipopolysaccharide (LPS)로 인한 nitric oxide (NO)의 생성과 inducible NO synthase (iNOS) 발현을 억제하였으며 HO-1 siRNA를 처리했을 때 ACDE가 iNOS의 발현을 억제하지 못하였다. 이상의 결과를 종합해보면, ACDE는 대식세포에서 PI3K/Akt 및 MAPK와 Nrf2 신호전달과정을 통해 HO-1의 발현을 유도함으로써 NO와 같은 염증매개물질의 생성을 억제한다는 것을 알 수 있다. 이러한 연구결과는 ACDE가 항염증제로 사용될 수 있음을 시사한다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.167-174
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• 2014

Tumor angiogenesis, growth and metastasis are three closely related processes. We therefore investigated the effects of barbigerone on all three in the B16F10 tumor model established in both zebrafish and mouse models, and explored underlying molecular mechanisms. In vitro, barbigerone inhibited B16F10 cell proliferation, survival, migration and invasion and suppressed human umbilical vascular endothelial cell migration, invasion and tube formation in concentration-dependent manners. In the transgenic zebrafish model, treatment with $10{\mu}M$ barbigerone remarkably inhibited angiogenesis and tumor-associated angiogenesis by reducing blood vessel development more than 90%. In vivo, barbigerone significantly suppressed angiogenesis as measured by H and E staining of matrigel plugs and CD31 staining of B16F10 melanoma tumors in C57BL/6 mice. Furthermore, it exhibited highly potent activity at inhibiting tumor growth and metastasis to the lung of B16F10 melanoma cells injected into C57BL/6 mice. Western blotting revealed that barbigerone inhibited phosphorylation of AKT, FAK and MAPK family members, including ERK, JNK, and p38 MAPKs, in B16F10 cells mainly through the MEK3/6/p38 MAPK signaling pathway. These findings suggested for the first time that barbigerone could inhibit tumor-angiogenesis, tumor growth and lung metastasis via downregulation of the MEK3/6/p38 MAPK signaling pathway. The findings support further investigation of barbigerone as a potential anti-cancer drug.

• 한국축산식품학회지
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• 제40권2호
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• pp.274-285
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• 2020

In this study, ovalbumin (OVA) hydrolysates were prepared using various proteolytic enzymes and the anti-inflammatory activities of the hydrolysates were determined. Also, the potential application of OVA as a functional food material was discussed. The effect of OVA hydrolysates on the inhibition of nitric oxide (NO) production was evaluated via the Griess reaction, and their effects on the expression of inducible NO synthase (inducible nitric oxide synthase, iNOS) were assessed using the quantitative real-time PCR and Western blotting. To determine the mechanism by which OVA hydrolysates activate macrophages, pathways associated with the mitogen-activated protein kinase (MAPK) signaling were evaluated. When the OVA hydrolysates were added to RAW 264.7 cells without lipopolysaccharide (LPS) stimulation, they did not affect the production of NO. However, both the OVA-Protex 6L hydrolysate (OHPT) and OVA-trypsin hydrolysate (OHT) inhibited NO production dose-dependently in LPS-stimulated RAW 264.7 cells. Especially, OHT showed a strong NO-inhibitory activity (62.35% at 2 mg/mL) and suppressed iNOS production and the mRNA expression for iNOS (p<0.05). Also, OHT treatment decreased the phosphorylation levels of Jun amino-terminal kinases (JNK) and extracellular signal-regulated kinases (ERK) in the MAPK signaling pathway. These findings suggested that OVA hydrolysates could be used as an anti-inflammatory agent that prevent the overproduction of NO.

• Journal of Ginseng Research
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• 제46권1호
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• pp.115-125
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• 2022

Background: Ginsenosides (GS) have potential value as cosmetic additives for prevention of skin photoaging. However, their protective mechanisms against skin barrier damage and their active monomeric constituents are unknown. Methods: GS monomer types and their relative proportions were identified. A UVB-irradiated BALB/c hairless mouse model was used to assess protective effects of GS components on skin epidermal thickness and transepidermal water loss (TEWL). Skin barrier function, reflected by filaggrin (FLG), involucrin (IVL), claudin-1 (Cldn-1), and aquaporin 3 (AQP3) levels and MAPK phosphorylation patterns, were analyzed in UVB-irradiated hairless mice or HaCaT cells. Results: Total GS monomeric content detected by UPLC was 85.45% and was largely attributed to 17 main monomers that included Re (16.73%), Rd (13.36%), and Rg1 (13.38%). In hairless mice, GS ameliorated UVB-induced epidermal barrier dysfunction manifesting as increased epidermal thickness, increased TEWL, and decreased stratum corneum water content without weight change. Furthermore, GS treatment of UVB-irradiated mice restored protein expression levels and epidermal tissue distributions of FLG, IVL, Cldn-1, and AQP3, with consistent mRNA and protein expression results obtained in UVB-irradiated HaCaT cells (except for unchanging Cldn-1 expression). Mechanistically, GS inhibited JNK, p38, and ERK phosphorylation in UVB-irradiated HaCaT cells, with a mixture of Rg2, Rg3, Rk3, F2, Rd, and Rb3 providing the same protective MAPK pathway inhibition-associated upregulation of IVL and AQP3 expression as provided by intact GS treatment. Conclusion: GS protection against UVB-irradiated skin barrier damage depends on activities of six ginsenoside monomeric constituents that inhibit the MAPK signaling pathway.

• Journal of Ginseng Research
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• 제47권3호
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• pp.420-428
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• 2023

Background: Ginsenoside F2 (GF2), a minor component of Panax ginseng, has been reported to possess a wide variety of pharmacological activities. However, its effects on glucose metabolism have not yet been reported. Here, we investigated the underlying signaling pathways involved in its effects on hepatic glucose. Methods: HepG2 cells were used to establish insulin-resistant (IR) model and treated with GF2. Cell viability and glucose uptake-related genes were also examined by real-time PCR and immunoblots. Results: Cell viability assays showed that GF2 up to 50 μM did not affect normal and IR-HepG2 cell viability. GF2 reduced oxidative stress by inhibiting phosphorylation of the mitogen-activated protein kinases (MAPK) signaling components such as c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (ERK1/2), and p38 MAPK, and reducing the nuclear translocation of NF-κB. Furthermore, GF2 activated PI3K/AKT signaling, upregulated the levels of glucose transporter 2 (GLUT-2) and GLUT-4 in IR-HepG2 cells, and promoted glucose absorption. At the same time, GF2 reduced phosphoenolpyruvate carboxykinase and glucose-6-phosphatase expression as well as inhibiting gluconeogenesis. Conclusion: Overall, GF2 improved glucose metabolism disorders by reducing cellular oxidative stress in IR-HepG2 cells via MAPK signaling, participating in the PI3K/AKT/GSK-3β signaling pathway, promoting glycogen synthesis, and inhibiting gluconeogenesis.

• 생명과학회지
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• 제32권11호
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• pp.833-840
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• 2022

만성 염증은 종양의 발생 및 진행과 밀접하게 연관되어 있다. 핵인자 kappa B (NF-κB)는 5개의 전사인자로 구성되며 염증 반응에 필수적인 역할을 한다. 다양한 암에서 NF-κB의 조절장애가 보고되고 있으며 NF-κB 조절이 암 치료에 있어 핵심 표적이 된다. 본 연구에서는 CDC Like Kinase 3 (CLK3)를 NF-κB 신호전달 경로를 조절하는 새로운 키네이스임을 확인하였다. 우리는 CLK3가 정규 및 비정규 NF-κB 신호전달경로를 억제하는 것을 밝혔다. CLK3 과발현 또는 knock-down 세포주를 이용한 루시퍼레이즈 분석 결과, 이 키네이스는 TNFα와 PMA가 유도하는 정규 NF-κB 신호전달경로 활성을 억제하였다. 또한 CLK3 과발현은 잘 알려진 비정규 NF-κB 신호경로 유도제인 NF-κB-inducing kinase (NIK) 또는 CD40에 의한 NF-κB 활성을 저해하였다. 추가적으로 CLK3의 NF-κB 신호전달 저해기전을 설명하고자 TNFα 처리 후 웨스턴 블롯 분석으로 이 키네이스 영향권 내에 있는 NF-κB 신호경로 분자들을 식별하였다. 그 결과 CLK3가 TAK1, IKKα/α, p65, IκBα 및 ERK1/2-MAPK의 인산화/활성화를 저해하여 TNFα 처리로 유도된 NF-κB 및 MAPK 신호경로를 모두 억제함을 밝혔다. 앞으로의 연구는 CLK3가 정규 및 비정규 NF-κB 경로를 억제하는 기작을 밝히는데 초점을 맞출 것이다. 위 연구 결과들을 토대로 CLK3가 NF-κB 신호전달경로의 새로운 음성 조절자로써 기능함을 제시하였다.

• 생명과학회지
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• 제33권6호
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• pp.471-480
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• 2023

본 연구에서는 LPS로 자극된 RAW 264.7 세포에서 생물전환 루모라고사리 추출물(B-RAE)의 항염증 효과 및 작용기전을 연구하였다. B-RAE의 총 폴리페놀과 총 플라보노이드 함량을 측정한 결과 379.26±7.77 mg/g과 50.85±3.08 mg/g으로 각각 나타났다. B-RAE의 항산화효과를 측정한 결과 DPPH, ABTS, superoxide anion radical을 농도의존적으로 소거하는 것으로 확인되었다. 또한, B-RAE는 세포생존에 영향을 미치지 않으면서 NO 생성을 처리 농도의존적으로 저해하였다. 전염증성 사이토카인(TNF-α, IL-1β, IL-6) 발현에 미치는 영향을 정량적 실시간 PCR로 측정한 결과 전염증성 사이토카인의 mRNA발현량을 LPS 처리군과 비교하여 B-RAE 처리 농도 의존적으로 유의성 있게 감소시키는 것으로 나타났다. 염증 관련 단백질(iNOS, COX-2)의 발현 및 전사인자인 NF-κB와 MAPK 신호경로 단백질의 인산화에 미치는 영향을 Western blot분석으로 평가하였다. 그 결과 LPS 처리에 의하여 증가된 iNOS와 COX-2의 발현을 유의적으로 억제하였다. 또한, LPS 처리에 의하여 증가된 NF-κB와 IκB의 인산화가 B-RAE 처리에 의하여 감소되는 것으로 나타났다. MAPK 신호경로 단백질의 인산화에 미치는 영향을 측정한 결과 ERK와 p38 MAPK 단백질의 인산화는 농도의존적으로 감소하는 것으로 나타났으나 JNK의 인산화는 증가하는 것으로 나타났다. 따라서 이러한 B-RAE의 항염증 효과는 높은 항산화 활성, iNOS와 COX-2 발현 억제를 통한 NO 생성 억제, NF-κB경로 저해, MAPK 신호경로 조절 및 전염증성 사이토카인 발현 저해에 의해 가능하다는 것을 제시한다.

• 대한본초학회지
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• 제21권1호
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• pp.33-42
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• 2006

Objectives : The purpose of this study was to investigate the anticancer effects of Bodusan (BDS) on SNU-1 cells, a human gastric cancer cell line. Methods : To study the cytotoxic effect of BDS on SNU-1 cells, the cells were treated with various concentrations of BDS and then cell viability was determined by XTT reduction method and trypan blue exclusion assay. The typical signs of apoptosis, was examined by western blot analysis. BDS-induced MAPK activation was also examined by Western blot for phosphorylated ERK and p38. Results : BDS reduced proliferation of SNU-1 cells in a dose-dependent manner and decreased procaspase 3 level in a dose-dependent manner and induced the clevage of PARP at concentration > 500 ${\mu}g/ml$. BDS also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol and reducing the level of anti-apoptotic Bcl-2. BDS significantly decreased ERK phosphorylation and increased p38 phosphorylation in a dose-dependent manner. Futhermore, BDS treatment up-regulated p53 and p21waf expression in a dose-dependent manner. Conclusion : BDS-induced apoptosis is MAP kinase-dependent apoptoric pathway and arrested SNU-1 cells at the G0/G1 of cell cycle. These results suggest that BDS is potentially useful as a chemotherapeutic agent in human gastric cancer.

• BMB Reports
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• 제42권3호
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• pp.148-153
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• 2009

Pyrrolidine dithiocarbamate (PDTC) is a stable anti-oxidant or pro-oxidant, depending on the situation, and it is widely used to inhibit the activation of NF-${\kappa}B$. We recently reported that PDTC activates the MIP-2 gene in a NF-${\kappa}B$-independent and c-Jun-dependent manner in macrophage cells. In this work, we found that PDTC activates signal transduction pathways in mouse ES cells. Among the three different mitogen-activated protein kinase (MAPK) pathways, including the extracellular-signal-regulated kinase (ERK), p38 MAP kinase, and stress-activated protein kinase (SAPK)/Jun N-terminal kinase (JNK) pathways, only the ERK pathway was significantly activated in mouse ES cells after stimulation with PDTC. Additionally, we observed a synergistic activation of ERK and induction of c-Fos after stimulation with PDTC in the presence of mouse embryonic fibroblast (MEF) conditioned medium. In contrast, another NF-${\kappa}B$ inhibitor, BMS-345541, did not activate the MAP kinase pathways or induce expression of c-Fos. These results suggest that changes in the presence of the NF-${\kappa}B$ inhibitor PDTC should be carefully considered when it used with mouse ES cells.

• 한국식품영양과학회지
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• 제46권7호
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• pp.896-902
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• 2017

본 연구는 감귤 콤부차의 산업화를 위한 발효 균주 표준화를 위하여 콤부차에서 분리된 3가지 균주(Gluconacetobacter xylinus, Gluconacetobacter medellinensis, Gluconobacter oxydans)를 이용한 감귤 발효액(CK-MOX)의 기능적 특성을 탐색하고자 하였다. CK-MOX 제조 후 15일간 3일 마다 샘플링을 하였으며, 발효 기간에 따른 pH, 산도, 항산화 능력을 평가하였다. 발효에 따라 pH는 감소하였고, 산도는 증가하였다. DPPH, ABTS 라디칼 소거능, ORAC assay를 통한 항산화 능력 측정 결과 발효에 따라 항산화 능력이 향상하는 것으로 나타났으며, 방광암 세포주(EJ 세포)의 생존 억제 및 이동 억제 효과가 있는 것으로 나타났다. 특히 CK-MOX로 유도된 EJ 세포의 사멸에 MAPK pathway의 중추적인 역할을 하는 것으로 알려진 ERK의 발현이 깊이 관여하는 것으로 나타났다.