• 대한화장품학회지
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• 제33권3호
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• pp.203-208
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• 2007

천연물로부터 새로운 미백 화장품 소재를 개발하기 위해 참나물(P. brachycarpa)을 선택하여 B16 멜라노마 세포에서 tyrosinase 활성 저해 효과, melanin 생성 저해 효과를 측정하였다. 먼저 참나물추출물의 4가지 극성별 용매 분획화 실험을 실시하였으며, 항산화효과와 티로시나아제 저해효과를 측정하였다. 참나물의 미백 활성 메카니즘을 알아보기 위해 Western blotting과 RT-PCR을 이용하여 tyrosinase, TRP-1, TRP-2의 단백질 발현과 mRNA 변화를 연구하였다. 또한, HaCaT 각질형성세포에서 UVB 조사 후 엔도세린-1(ET-1)의 발현은 사람 ET-1 항체를 이용하여 quantitative enzyme immunoassay(EIA)로 측정하였다. 그 결과 참나물추출물과 4가지 분획(hexane, EtOAc, butanol and aqueous)은 100 ${\mu}g/mL$ 농도에서 각각 87.2, 2.5, 97.2, 80.5, 49.8%의 프리라디칼 소거효과를 나타내었으며, tyrosinase 저해효과는 100 ${\mu}g/mL$ 농도에서 각각 18.3, 15.1, 55.4, 13.1, 0%로 나타났다. 각 극성별 분획 중 EtOAc 분획 100 ${\mu}g/mL$ 농도에서 58% 이상의 가장 우수한 멜라닌 생성 저해 효과가 나타났다. 참나물 EtOAc 분획은 B16 멜라노마 세포에서 티로시나아제 활성 및 발현을 모두 저해하였으며, RT-PCR 결과에서도 tyrosinase, TRP-1의 mRNA 발현 저해효과가 우수하게 나타났다. 또한 HaCaT 각질형성세포에서 UVB 조사 후 생성된 엔도세린-1의 생성 실험에서도 참나물 EtOAc 분획 $12.5{\sim}50{\mu}g/mL$ 농도에서 엔도세린-1의 생성이 컨트롤에 비해 40% 정도로 우수하게 저해되었다. 결론적으로 참나물추출물은 멜라닌 합성과정에서 우수한 tyrosinase 저해효과와 엔도세린-1 발현저해효과를 가지는 새로운 천연 미백 소재로 적용될 수 있을 것이라 기대된다.

• 미생물학회지
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• 제33권1호
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• pp.1-6
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• 1997

5-Enolpyruvylshikimate 3-phosphate(EPSP) synthase는 방향족 아미노산을 생합성하는 shikimate phathway의 6번째 효소로 광범위 제초제인 glyphosate의 target enzyme이다. 본 연구에서는, glyphosate에 저해를 받지 않는 EPSP synthase를 개발하고자 하는 연구의 한 단계로서, 우선, EPSP synthase를 대량 발현시킬수 있는 expression vector인 pET-25b를 사용하여 발현시킨 다음, 발현된 효소가 periplasmic space로 transport되는지 또 발현된 단백질이 효소 활성을 가지고 있는지 확인하고자 하였다. 그 결과, pelB leader를 앞에 붙여 발현시킨 EPSP synthase는 periplasmic space로 제대로 transport되며, 단백질 생산 및 periplasmic space로의 수송은 induction 온도에 의해 크게 좌우된다는 것을 관찰하였다. Periplasmic space로 수송되는 EPSP synthase의 양은 $34^{\circ}C$에서 induction시켰을 때 가장 많은 것으로 나타났다. 한편, pET-25b를 이용하여 발현시킨 EPSP synthase는 C-terminal 부위에 HSV-tag, His-tag등 26개 아미노산이 더 있는 상태로 만들어지는데, His-tag은 $Ni^{2+}$-affinity chromatography를 통한 정제에, HSV-tag은 Western blotting을 통한 detection에 각각 이용할 수 있다. 또한, 이와 같이 발현된 recombinant EPSP synthase는 phosphocellulose resin에 결합하였다가 기질인 shikimate 3-phosphate와 phosphoenolpyruvate에 의해 elution되며, glyphosate에 의해 저해되는등 wildtype효소와 같은 효소 특성을 보였다.

• The Korean Journal of Physiology and Pharmacology
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• 제4권3호
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• pp.185-195
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• 2000

Vascular diseases are significant complications of diabetes mellitus (DM), and the endothelial cells may play a pivotal role in the development of vascular disease in DM. Endothelin-1 (ET-1) released from endothelium is a potent vasoconstrictor peptide and circulating level of ET-1 is increased in a variety of disease states. The purpose of this study was to determine the changes of responsiveness to ET-1 in DM, and we experimented on the changes in the ET-1-induced contraction, levels of nitrite and lipid peroxidation, and ET-1 immunoreactivity in aorta from streptozotocin-induced DM rats. DM was induced by single injection of streptozotocin (55 mg/kg, i.p.). The immunoreactive ET-1 levels in endothelial layer of thoracic aorta were much higher in DM rats than control rats. Nitrite in tissue homogenate was decreased and plasma nitrite was increased in DM rats. Malondialdehyde (MDA) was significantly increased in DM rats and cGMP was not significantly different between control and DM rats. ET-1 produced concentration- dependent contractile responses that are significantly attenuated in DM rats compared to controls. In the presence of selective $ET_A$ receptor antagonist BQ610, the maximum contraction was decreased and the concentration ratios for BQ610 yielded $pA_2$ values of 7.3 (slope, 0.65) in control rats, whereas BQ610 had no antagonistic effect on ET-1-induced contraction in DM rats. However, pretreatment with BQ788, an $ET_B$ receptor antagonist, maximum response was decreased and the dose-response curves for ET-1 were shifted to the right in both groups and $pA_2$ values were 7.9 and 7.7 (slope, 1.05 in control and DM rats), respectively. IRL 1620 and sarafotoxin S6c, $ET_B$ agonists, induced relaxation in control rats but not in DM rats. These results indicate that endothelial cell dysfunction and enhanced immunoreactivity of ET-1 have been found in DM rat and ET-1-induced contraction was attenuated in DM rat. These attenuated responses might be at least in part caused by the alteration of $ET_A$ receptor properties (e.g. desensitization), and partly related with an alteration in intracellular mechanism for contraction to ET-1.

• Journal of Applied Biological Chemistry
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• 제57권2호
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• pp.165-170
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• 2014

덜꿩나무(Viburnum erosum) 줄기를 실온에서 80% MeOH 수용액으로 추출하고 농축하였다. 이 추출물을 EtOAC, n-BuOH, 및 $H_2O$ 분획으로 나누었다. 이 중 EtOAc및 n-BuOH 분획에 대하여 $SiO_2$, ODS 및 Sephadex LH-20 column chromatography를 반복 수행하여 4종의 화합물을 분리, 정제하였다. 분리한 화합물의 구조는 NMR, IR, 및 MS data를 해석하여, betulinic aldehyde (1), koaburside (2), (6R,7E,9R)-9-hydroxymegastigma-4,7-dien-3-one-9-O-${\beta}$-D-glucopyranoside (3), 그리고 byzantionoside B (4)로 구조를 결정하였다. 분리된 화합물 모두 덜꿩나무에서는 처음으로 분리되었다.

• 한방안이비인후피부과학회지
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• 제17권1호
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• pp.131-142
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• 2004

The purpose of this research was to investigate cytotoxity of Sunbanhwalmyungeum extract. Cytotoxity was determined by MTT assay method. After tumor cell lines(G361, BI6F10, MDA, A549) transplantation, the extracts of SunBangHwalMyungEum and its composition oriental medicines were administered, cytotoxity was measured by absorbance. The results were obtained as follows. 1. Sunbanhwalmyungeum extract and its composition oriental medicines extracts showed the concentration was higher, the more cytotoxity increased. 2. Both water and ethanol extracts of Sunbanhwalmyungeum showed excellent cytotoxity against G361, B16F10, MDA, A549 and high cytotoxity over 80$\%$ against G361, B16F10, MDA except A549 at the concentration of 1000ppm. 3. In water extract, Rhei Radix et Rhizoma, Gleditsiae Spina, Trichosanthis Radix, Glycyrrhizae Radix, Ledebouriellae Radix showed excellent cytotoxity. In ethanol extract, Gleditsiae Spina, Citri Pericarpium, Trichosanthis Radix, Paeoniae Radix Rubra, Myrrha showed excellent cytotoxity. 4. Rhei Radix et Rhizoma, Gleditsiae Spina, Trichosanthis Radix, Paeoniae Radix Rubra showed high cytotoxity in both water and ethanol extrats. 5. In water extract, Glycyrrhizae Radix, Ledebouriellae Radix, Myrrha showed high cytotoxity against A361, Lonicerae Flos, Olibanum, Fritillariae cirrhosae Bulbus, Paeoniae Radix Rubra, Manitis Squama against B16F10, Paeoniae Radix Rubra, Manitis Squama against MDA, Rhei Radix et Rhizoma, Angelicae gigantis Radix against A549. 6. In ethanol extract, Lonicerae Flos, Trichosanthis Radix showed high cytotoxity against G361, Rhei Radix et Rhizoma, Angelicae gigantis Radix, Gleditsiae Spina, Olibanum, Angelicae dahuricae Radix, Fritillariae cirrhosae Bulbus, Paeoniae Radix Rubra, Glycyrrhizae Radix, Ledebouriellae Radix, Myrrha against B16F10, Rhei Radix et Rhizoma, Manitis Squama against MDA, Citri Pericarpium, Manitis Squama against A549.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.73-86
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• 2003

Endothelins secreted from keratinocytes are intrinsic madiators for human melanocytes in UVB-induced pigmentation. Antimelanogenic ingredient, 1,2-Ο-diferulylglycerol(SM709) isolated from bamboo extract inhibited the melanin synthesis of Bl6F10 melanoma cells by 62%. To understand the cellular mechanism of antimelanogenic activity of SM709 in human melanocytes, the effects of SM709 on the ET-l-induced $Ca^{2+}$ mobilization were investigated. ET-l receptors in human melanocytes were characterized by using specific antagonist and found that ET-l increased intracellular $Ca^{2+}$ by activating ET-B receptor. SM709 completely blocked the ET-l-induced intracellular $Ca^{2+}$ increase and its inhibitory effect showed dose- and time- dependent manners. To investigate the role of SM709 on intracellular $Ca^{2+}$ store, when the $Ca^{2+}$ store was partially depleted by thapsigargin; a specific inhibitor of ER-type $Ca^{2+}$-ATPase, caffeine-induced $Ca^{2+}$ mobilization did not changed in the presence or absence of SM709, suggesting that SM709 has no effect on the $Ca^{2+}$ store. It is known that LPA receptor and P$_2$ receptor are linked to InsP$_3$ second messenger system. When these receptors in melanocytes were activated by LPA and ATP, the intracellular $Ca^{2+}$ signaling was observed even in the presence of SM709. From the above results, it can be suggested that SM709 has an antimelanogenic activity by antagonizing the ET-B receptor, resulting in subsequent intracellular $Ca^{2+}$ signaling, in UV induced pigmentation.nduced pigmentation.

• Bulletin of the Korean Chemical Society
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• 제13권1호
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• pp.80-83
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• 1992

The secondary nitrogen donors of the Ni(II) complexes of macrotricyclic ligands 8-methyl-1,3,6,8,10,13,15-heptaazatricyclo $[13.1.1.1^{13,15}]$octadecane (A) and 1,3,6,9,11,14-hexaazatricyclo $[12.2.1.1^{6,9}]$octadecane (B) are N-alkylated and the Ni(II) complexes of $N-Me_2A,\;N-Et_2A,\;and\;N-Me_2B$ are obtained. The Ni(II) complexes of $N-Me_2A\;and\;N-Et_2A$ are stable in acidic aqueous solutions while that of $N-Me_2B$ decomposes relatively rapidly. The N-alkylation leads to the decrease in the ligand field strength as well as an anodic shift in both of the oxidation and the reduction potentials of the Ni(II) complexes.

• 대한한의학회지
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• 제40권2호
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• pp.35-50
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• 2019

Objectives: Orostachys japonicas (O. japonicus) has been known for its anti-tumor effect. In the present study, it was investigated whether O. japonicus EtOH extracts could induce apoptosis and autophagy which are part of the main mechanism related to anti-tumor effect in THP-1 cells. Methods: Cells were treated with various concentrations of O. japonicus EtOH extracts ($0-300{\mu}g/ml$) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay and apoptosis rate was examined by flow cytometry and ELISA assay. The mRNA expression of apoptosis-related genes (Bcl-2, Mcl-1, Survivin, Bax) and autophagy-related gene (mTOR) was evaluated using real-time PCR. The protein expression of Caspase-3, Akt, LC3 II, Beclin-1, Atg5, $NF-{\kappa}B$, p38, ERK was evaluated using western blot analysis. Results: O. japonicus EtOH extracts inhibited cell proliferation and apoptosis rate was increased in both flow cytometry and ELISA assay. Bcl-2, Mcl-1, Survivin (anti-apoptosis factors) mRNA expressions were decreased and Bax (pro-apoptosis factor) mRNA level was increased. mTOR mRNA expressions was decreased and LC3 II protein expressions was increased. Activation of $NF-{\kappa}B$ was decreased and phosphorylation of p38 was increased. Conclusion: O. japonicus is regarded to inhibit cell proliferation, to induce apoptosis and to regulate autophagy-related genes in THP-1 cells via $NF-{\kappa}B$ and p38 MAPK signaling pathway. This suggests O. japonicus could be an effective herb in treating acute myeloid leukemia.

• KSBB Journal
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• 제9권5호
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• pp.506-511
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• 1994

천연자원에서 향균성 물질을 찾고자 수종의 식용 식물을 검색하는 과정에서 쑥(Artemisia princeps var. orientalis)의 추출물이 항균활생이 비교적 높았다. 쑥의 메탄올 추출물을 겨l통분획한 ethyl acetate 분획물(EtOAc)은 agar diffusion법에셔 항균활성이 가장 높았으며, EtOAc로부터 분리된 화합물 중 0­C coumaric acid(200 -600ppm)는 B. subtilis(12.6 ~ -18.0mm)와 s. typmηwrium( 12.6-16.6mm)에 대하 여 우수한 항균활성을 나타내였고, 그 유도체인 p­C coumanc a디d는 약 1.2 -1.7배 정도의 항균활성을 증가시켰다 Coumaric acid의 3가지는 액체배양에 서 B. subtilis 증식응 모두 강하게 억제하였으며, S. typmmuriuη2, P. aeruginosa 및 s. aureus의 증식 억제에는 각각 0-, m- 및 p-coumaric a디d가 효과적 이었다. B. subtilis의 반고체 agar 배양에셔 pc coumaric acid의 MID는 $\100∼200mu\textrm{g}$/disk였다.

• 한국식품과학회지
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• 제29권1호
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• pp.156-160
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• 1997

식용버섯류들의 MAO 저해활성을 검색할 목적으로 몇가지 유기용매를 이용하여 추출분획을 조제하고 이들의 MAO에 대한 저해활성을 검색하였다. 검색된 3종 버섯류의 유기용매 추출물들은 영지버섯 bud의 $CHCl_3$ 층을 제외하고는 모두 MAO-A에 대한 저해활성이 나타나지 않았으나 영지 bud의 경우 $CHCl_3$ 층에서 비교적 강력한 MAO-A 저해활성을 나타내어 영지 bud의 항암활성이 영지의 항암활성과 비교하여 더욱 강력하다는 기존의 보고와 관련하여 아주 흥미로운 결과로 영지의 성분연구에 아주 중요한 단서를 제공해 줄 것으로 생각된다. 또한 검색된 3종 버섯류의 유기용매 추출물들은 영지 bud, 표고버섯의 EtOAc 층에서 각각 51.3, 55.9%로 역한 정도의 저해활성이 관찰되었으며 그 외의 다른 버섯류들에서는 전혀 MAO-B에 대한 저해활성이 확인되지 않았다. 또한 영지버섯의 경우, $CHCl_3$ 층, EtOAc 층, $H_2O$ 층 모두에서 아주 미약하나마 MAO-B에 대한 저해활성을 나타냈으나 영지 bud의 경우 $CHCl_3$ 층에서 전혀 MAO-B에 대한 저해활성을 나타내지 않고 EtOAc 층에서 약한 MAO-B에 대한 저해활성이 확인되어 MAO-A와 MAO-B에 대한 영지버섯의 작용성분이 다른 성분일 것으로 추측된다.