• 한국수산과학회지
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• 제51권2호
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• pp.127-134
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• 2018

Arteriosclerosis is the major cause of coronary artery and cerebrovascular disease, which are leading causes of death. Pro-inflammatory cytokines induce injury to vascular endothelial cells by increasing cell adhesion molecules, leading to vascular inflammation, a major risk factor for the development of arteriosclerosis. In the current study, we investigated the inhibitory effect of enzymatic hydrolysate from Japanese mud shrimp Upogebia major on the inflammation of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$)-stimulated human umbilical vein endothelial cells (HUVECs). We first evaluated the antioxidant and angiotensin I-converting enzyme (ACE) inhibitory activities of eight U. major enzymatic hydrolysates: alcalase, papain, ${\alpha}$-chymotrypsin (${\alpha}-Chy$), trypsin, pepsin, neutrase, protamex and flavourzyme. Of these, ${\alpha}-Chy$ exhibited potent antioxidant and ACE inhibitory activities. The ${\alpha}-Chy$ hydrolysate was fractionated by two ultrafiltration membranes of 3 and 10 kDa. The ${\alpha}-Chy$ hydrolysate of U. major and its molecular weight cut-off fractions resulted in a significant reduction in NO production and a decrease in cell adhesion molecules [vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and endothelial-selectin (E-selectin)] and pro-inflammatory cytokines [interleukin-6 (IL-6), interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1)] in $TNF-{\alpha}$-stimulated HUVECs. These results suggest that enzymatic hydrolysate from U. major can be used in the control and prevention of vascular inflammation and arteriosclerosis.

• 한국미생물·생명공학회지
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• 제48권1호
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• pp.79-89
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• 2020

본 연구는 단풍잎돼지풀 70% 에탄올 추출물의 항염증 활성을 검증하기 위해 수행되었다. 단풍잎돼지풀 70% 에탄올 추출물의 전자공여능 측정과 ABTS⁺ 라디칼 소거능을 측정한 결과, 1,000 ㎍/ml 농도에서 각각 84.1%와 92.5%의 효과를 나타냈고, 수렴활성 측정을 한 결과 1,000 ㎍/ml 농도에서 94.7%의 효과를 보였다. 단풍잎돼지풀 70% 에탄올 추출물의 항염증 효과를 측정하기 위해 lipopolysaccharide (LPS)로 유도된 RAW 264.7세포를 사용하여 효과를 측정하였다. 세포에서 단풍잎돼지풀 추출물의 세포독성을 측정하기 위해 MTT assay를 수행하였다. 그 결과, 500 ㎍/ml 농도에서 90% 이상의 생존율을 보였다. Nitric oxide 생산을 억제하는 효과를 측정한 결과, 단풍잎돼지풀 추출물에서 농도가 증가할수록 NO 생성이 감소되는 효과를 확인하였다. 단풍잎돼지풀 추출물의 단백질 발현효과를 western blot을 통해 25, 50, 100 ㎍/ml 농도에서 측정하였고, 양성 대조군으로 β-actin을 사용하였다. 그 결과, inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 단백질 발현효과는 100 ㎍/ml 농도에서 8.6%, 25.1%의 감소됨을 확인하였다. ERK1/2, p38, JNK와 Iκ-Bα의 단백질발현 효과는 인산화를 통해 확인하였고, 농도의존적으로 감소하였음을 확인하였다. mRNA 발현 억제 효과를 RT-PCR을 통해 25, 50, 100 ㎍/ml의 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, LPS로 유도된 대식세포에서 iNOS, COX-2, interleukin (IL)-1β, IL-6, TNF-α의 mRNA 발현 억제 효과는 농도가 증가할수록 발현이 감소됨을 확인하였다. 결론적으로 단풍잎돼지풀 추출물은 염증을 억제할 수 있는 가능성이 있는 항염증 소재의 가능성을 증명하였다.

• 한국자원식물학회지
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• 제28권5호
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• pp.568-573
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• 2015

본 연구에서는 장대나물(Arabis glabra)의 전초를 이용하여 세포독성 및 항염증 활성 효과를 평가하였다. 대식세포인 RAW264.7 cell에서 염증 매개 물질인 lipopo-lysacchride (LPS)로 염증을 유발시켜 nitric oxide (NO)와 prostaglandi nE₂ (PGE₂) 같은 염증 유발인자들의 억제효과를 확인하였다. 장대나물의 hexane과 chloroform 분획물의 염증 유발인자 억제 시 IC₅₀ value를 측정하였을 때 nitric oxide 및 prostaglandin E₂ 생성을 농도의존적으로 현저하게 저해하는 농도는 각각 21.0과 18.0 ㎍/㎖였다. 따라서 본 연구 결과는 장대나물의 hexane, chloroform과 같은 비극성용매 분획물들이 유의성 있는 항염증 효과를 나타내었으며, 이러한 효능은 예방의학적 가능성을 충분히 가지고 있기에 염증성질환의 예방을 위한 건강 기능성식품의 개발 가능성을 제시할 수 있을 것으로 기대된다. 또한 염증과 관련된 사이토카인 및 단백질 발현 메커니즘에 대한 추가적인 연구가 필요할 것으로 판단된다.

• 한국해양바이오학회지
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• 제12권1호
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• pp.29-39
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• 2020

In this study, we investigated the inhibitory potential of an ethanol extract of Carpomitra costata (EECC) (Stackhouse) Batters, a brown alga, against neuroinflammatory responses in lipopolysaccharide (LPS)-stimulated BV2 microglia. Our results showed that EECC significantly suppressed the LPS-induced secretion of pro-inflammatory mediators, including nitric oxide (NO) and prostaglandin E₂, with no significant cytotoxic effects. EECC also inhibited the LPS-induced expression of their regulatory enzymes, such as inducible NO synthase and cyclooxygenase-2. In addition, EECC downregulated the LPS-induced expression and production of the proinflammatory cytokines, tumor necrosis factor-α and interleukin-1β. In the mechanistic assessment of the antineuroinflammatory effects, EECC was found to inhibit the nuclear translocation and DNA binding of nuclear factor-kappa B (NF-κB) by disrupting the degradation of the κB-α inhibitor in the cytoplasm. Moreover, EECC effectively suppressed the enhanced expression of Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88, as well as the binding of LPS to TLR4 in LPS-treated BV2 cells. Furthermore, EECC markedly reduced the LPS-induced generation of reactive oxygen species (ROS), demonstrating a strong antioxidative effect. Collectively, these results suggest that EECC repressed LPS-mediated inflammatory action in the BV2 microglia through the inactivation of NF-κB signaling by antagonizing TLR4 and/or preventing ROS accumulation. While further studies are needed to fully understand the anti-inflammatory effects associated with the antioxidant activity of EECC, the current findings suggest that EECC has a potential advantage in inhibiting the onset and treatment of neuroinflammatory diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.315-320
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• 2013

Here, we show that radicicol, a fungal antibiotic, resulted in marked inhibition of inducible nitric oxide synthase (iNOS) transcription by the pancreatic beta cell line MIN6N8a in response to cytokine mixture (CM: TNF-${\alpha}$, IFN-${\gamma}$, and IL-$1{\beta}$). Treatment of MIN6N8a cells with radicicol inhibited CM-stimulated activation of NF-${\kappa}B$/Rel, which plays a critical role in iNOS transcription, in a dose-related manner. Nitrite production in the presence of PD98059, a specific inhibitor of the extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) pathway, was dramatically diminished, suggesting that the ERK1/2 pathway is involved in CM-induced iNOS expression. In contrast, SB203580, a specific inhibitor of p38, had no effect on nitrite generation. Collectively, this series of experiments indicates that radicicol inhibits iNOS gene expression by blocking ERK1/2 signaling. Due to the critical role that NO release plays in mediating destruction of pancreatic beta cells, the inhibitory effects of radicicol on iNOS expression suggest that radicicol may represent a useful anti-diabetic activity.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.385-393
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• 2008

As an attempt to search for bioactive natural products exerting anti-inflammatory activity, we evaluated the effects of the methanol extract of Polytrichum commune Hedw (PCM) (Polytrichaceae) on lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines release in murine macrophage cell line RAW 264.7. PCM potently inhibits the production of NO, $PGE_2$, tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Consistent with these results, PCM also concentration-dependently inhibited LPS-induced inducible NO synthase (iNOS) and cyclooxygase (COX)-2 at the protein levels, and iNOS, COX-2, TNF-$\alpha$ and IL-6 at the mRNA levels without an appreciable cytotoxic effect on RAW 264.7 macrophag cells. Furthermore, PCM inhibited LPS-induced nuclear factor-kappa B (NF-$\kappa$B) activation as determined by NF-$\kappa$B reporter gene assay, and this inhibition was associated with a decrease in the nuclear translocation of p65 and p50 NF-$\kappa$B. Taken together, these results suggest that PCM may play an anti-inflammatory role in LPS-stimulated RAW 264.7 macrophages through the inhibitory regulation of iNOS, COX-2, TNF-$\alpha$ and IL-6 via NF-$\kappa$B inactivation.

• 동의생리병리학회지
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• 제25권2호
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• pp.227-233
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• 2011

Water Fermented and Ethanol Fermented Extracts from Rhei Radix et Rhizoma exhibit potent anti-inflammatory activity with an unknown mechanism. To elucidate the molecular mechanisms of W-FR and E-FR on pharmacological and biochemical actions in inflammation, we examined the effect of W-FR and E-FR on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. The investigation focused on whether FR inhibited nitric oxide (NO), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6) and the expressions of iNOS, COX-2 in LPS-stimulated RAW 264.7 cells. We found that FR inhibited LPS-induced NO, TNF-${\alpha}$ and IL-6 productions as well as the expressions of iNOS and COX-2. These results suggest that W-FR and E-FR have inhibitory effects on LPS-induced TNF-${\alpha}$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage.

• 생약학회지
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• 제53권4호
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• pp.192-201
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• 2022

This study aimed to compare bioactivities of Centella asiatica (CA) cultivated in smart farms and fields. Component analysis, cell viability, anti-inflammatory activity, neuroprotection activity, and antioxidant activity were examined with 70% ethanol extracts of CA cultivated in smart farm (SEE) and field (FEE), respectively. Asiaticoside was analyzed by high performance liquid chromatography (HPLC) and as a result, SEE had more asiaticoside content than FEE. After treatment of RAW 264.7 cells with SEE and FEE, there was no cytotoxicity within the treated concentrations. SEE and FEE showed nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin (IL)-6 inhibitory activities in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Moreover, SEE inhibited more NO, TNF-α, and IL-6 production levels than FEE. SEE and FEE reversed the H₂O₂-induced SH-SY5Y cell death. Especially, SEE was more effective in changing the H₂O₂-induced SH-SY5Y cell death than FEE. The antioxidant activity was confirmed by various methods such as total phenol content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and superoxide dismutase (SOD). As a result, SEE showed the most potent antioxidant activities about TPC, DPPH, and SOD methods. This study suggested that SEE has higher bioactivities such as effect of anti-inflammation, neuroprotection, and antioxidation than FEE.

• 한국응용과학기술학회지
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• 제39권2호
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• pp.303-313
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• 2022

본 연구는 장흥지역에서 자생 및 재배한 총 3가지 식물을 수증기 증류 추출(편백, 석창포, 라벤더)을 통해 얻은 각 천연 에센셜 오일에 대한 GC-MSD 향기 성분 분석 후 추출된 오일들과 시판 오일(클라리세이지, 시나몬, 프랑킨센스)을 조합하여 세포독성, 항산화, 항염, 미백 효과를 확인하여 화장품 소재로서의 가능성을 확인하였다. 향기 성분 분석 결과 편백 오일의 주요 성분으로는 Sabinene, 석창포 오일은 Asarone, 라벤더 오일은 L-Linalool이 동정되었다. 6종 에센셜 조합 오일의 세포독성이 확인되지 않은 100 𝜇L/mL의 농도에서 NO 생성은 27.76% 억제, DPPH 라디칼 소거능은 99.69%, ABTS 라디칼 소거능은 94.66%, 티로시나아제 저해활성은 55.9%를 보여 기능성 화장품 원료로서 유용하게 활용할 수 있는 가능성을 제시하였다.

• 생명과학회지
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• 제27권5호
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• pp.553-560
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• 2017

마늘은 전통적으로 cycloalliin을 포함한 기능성 식품으로 널리 알려져 있다. 본 연구를 통해 혈관계에서 발효마늘추출물이 어떤 효능을 갖는지 알아보았다. 먼저, 발효마늘추출물이 내피세포의 단핵구 부착을 억제하는 것을 관찰하였다. 농도 변화에 따른 실험결과, 발효마늘추출물을 0.1 또는 $1{\mu}g/ml$ 농도로 전처리 하면, LPS에 의하여 촉발되는 소 대동맥 내피세포에 달라붙는 THP-1의 부착이 억제되는 것을 확인하였다. 단핵구의 내피세포 부착이 염증반응 초기단계에 진행되는 과학적 근거를 고려하면 이와 같은 결과는 발효마늘추출물이 혈관 염증을 저해한다고 볼 수 있다. 또한 세포신호전달물질의 활성변화를 관찰한 결과, 발효마늘추출물이 소 대동맥 내피세포에서 eNOS와 Akt의 활성을 유도함을 알았다. eNOS의 인산화 반응은 발효마늘추출물을 처리한지 10분 후에 최대로 상승되었으며, 이와 유사하게 Akt도 발효마늘추출물을 처리하고 10분 후에서 인산화 반응이 최대로 상승되었다. 산화질소 양 변화 측정에서도 발효마늘추출물은 산화질소의 생성을 상승시켰다. 한편, 산화질소 생성을 저해하는 억제제 처리는 발효마늘추출물에 의해 나타나는 세포부착 저해능력을 감소시켰다. eNOS와 Akt를 매개로 하는 LPS 유도 세포 부착 정도가 발효마늘추출물에 의하여 억제된다는 것은 산화질소가 세포부착에 중요 매개물임을 의미한다. 결론적으로, 발효마늘추출물은 염증관련 질환을 예방하는 데에 중요한 역할을 한다고 할 수 있다.