• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.3
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• pp.203-208
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• 2007

To develop a new whitening agent for cosmetics from natural products, Pimpinella brachcarpa was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Crude ethanolic extract of P. brachycarpa and its four fractions-hexane, ethyl acetate(EtOAc), butanol and aqueous were evaluated for antioxidative effects and tyrosinase inhibitory activity. To elucidate the mechanism of active compounds of P. brachycarpa, we investigated the changes in protein level of tyrosinase, TRP-1 and TRP-2 using Western blotting and the changes in mRNA level of tyrosinase using RT-PCR technique. Following UV irradiation, expression of ET-1 in HaCaT keratinocytes was measured by quantitative enzyme immunoassay(EIA) using human ET-1 antibody. Crude ethanolic extract of P. brachycarpa and its four fractions-hexane, EtOAc, butanol and aqueous had free radical scavenging effect by 87.2, 2.5, 97.2, 80.5, 49.8% at 100 ${\mu}g/mL$ and tyrosinase inhibitory effect by 18.3, 15.1, 55.4, 13.1, 0 % at 100 ${\mu}g/mL$. P. brachycarpa EtOAc fraction significantly inhibited melanin production in B16 melanoma cells. Treatment with P. brachycarpa extract for 72 h suppressed the biosynthesis of melanin up to 58 % at 100 ${\mu}g/mL$. Especially, the EtOAc fraction of P. brachycarpa reduced the tyrosinase activity and tyrosinase expression in B16 melanoma cells in a dose-dependent manner. mRNA levels of tyrosinase and TRP-1 were markedly reduced by the EtOAc fraction of P. brachycarpa. Moreover, at the concentrations of $12.5{\sim}50{\mu}g/mL$ of the fraction, the production of UV-induced ET-1 in HaCaT keratinocytes(24 h after 8 $mJ/cm^2$ UVB irradiation) was reduced about 40%(p<0.05). P. brachycarpa could be used as a new natural skin-whitening agent due to the inhibitory effect of on melanin biosynthesis and endothelin-1 expression.

• Korean Journal of Microbiology
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• v.33 no.1
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• pp.1-6
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• 1997

5-Enolpyruvylshikimate 3-phosphate(EPSP) synthase is the sixth enzyme of the shikimate pathway that synthesizes aromatic amino acids. The enzyme is a primary target for the glyphos'lte which is a broad-spectrum and environmetally safe herbicide. As a first step toward development of glyphpsate-resistant EPSP synthase, the EPSP synthase gene(aroA) was amplified by polymerase chain reaction and cluned into pET-25b vector. In this construct. designated pET-aro, the aroA gene is expressed under control of strong T7 promoter. and the EPSP synthase is produced as a fusion protein with pelB leader at N-terminus and HSV-tag and His-tag at C-terminus. When the pET-aro clone was induced to produce the enzyme, it was found that the EPSP synthase was successfully exported to peri plasmic space. The periplasmic transport was greatly dependent on the induction temperatures. Among the induction temperatures examined($25^{\circ}C$, $30^{\circ}C$, $34^{\circ}C$ and $37^{\circ}C$). induction at $34^{\circ}C$ gave rise to maximal periplasmic transport. The recomhinant EPSP synthase could have been purified hy $Ni^{2+}$ -affinity chromatography using the His-tag. and detected hy anti-HSV -tag antibody. The recombinant EPSP synthase also hound to phosphocellulose resin and was eluted hy shikimate 3-phosphate and phosphoenolpyruvate. as expected. The recombinant EPSP synthase purified from phosphocellulose resin showed typical EPSP synthase activity.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.3
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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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• v.57 no.2
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• pp.165-170
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• 2014

The stems of Viburnum erosum were extracted with 80% MeOH. The concentrated extract was partitioned with EtOAc, n-BuOH, and $H_2O$. From the EtOAc and n-BuOH fractions, four compounds were isolated through the repeated $SiO_2$, octadecyl silica gel, and Sephadex LH-20 column chromatographies. Based on NMR, MS, and IR spectroscopic data, the chemical structures were determined as betulinic aldehyde (1), koaburside (2), (6R,7E,9R)-9-hydroxymegastigma-4,7-dien-3-one-9-O-${\beta}$-D-glucopyranoside (3), and byzantionoside B (4). All the compounds were isolated for the first time from the stems of Viburnum erosum.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.17 no.1
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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.

• Proceedings of the SCSK Conference
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• 2003.09b
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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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• v.13 no.1
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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.

• The Journal of Korean Medicine
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• v.40 no.2
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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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• v.9 no.5
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• pp.506-511
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• 1994

Antimicrobial activity of methanol extract and fraction from mugwort leaves(Artemisia princeps val. orientalis) was investigated for the screening of natural antiwucroblal components. By using agar diffusion method, ethyl acetate(EtOAc) layer fractionated from methanol extract of mugwort leaves showed the highest inhibitory effects against tested microorganisms. The ortho-coumaric acid(200∼600ppm) isolated from EtOAc layer showed strong antibacterial activities for Bacillus subtilis and Salmonella typhimurium. As derivatives of o-coumaric acid, antibacterial activity of para-coumaric acid was 1.2∼1.7 fold higher than that of o-coumaric acid. Three types of coumaric acids strong inhibited the growth of B. subtilis in the culture medium. Growth of S. tyhimurium, P. aeruginosa and S. aureus were effectively inhibited by o-, m- and p-coumaric acids, respectively. Minimum inhibitory dose of p-coumaric acid for B. subtilis was $\100∼200mu\textrm{g}$/paper disk.

• Korean Journal of Food Science and Technology
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• v.29 no.1
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• pp.156-160
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• 1997

The monoamine oxidase (MAO, EC 1.4.3.4) plays a central role in the metabolism of many amines including the neurotransmitter monoamines. MAO is a flavoprotein found exclusively in the mitochondrial outer membrane, occuring in the MAO-A and MAO-B subtypes. MAO-A deaminates serotonin and noradrenaline much better than phenethylamine (PEA) or benzylamine (BA), and is preferentially inhibited by clorgyline, whereas MAO-B prefers PEA and BA as substrates and is preferentially inhibited by deprenyl. MAO inhibitors were among the first drugs used in the treatment of depression, and it is known to be the inhibition of MAO-A which is important for the antidepressant effect of MAO inhibitors. For the purpose of evaluating MAO inhibitory activities from natural resources, three kinds of edible mushrooms were screened by tracing the inhibitory activities against rat brain mitochondrial MAO-A, utilizing serotonin as a substrate and rat liver mitochondrial MAO-B utilizing benzylamine as a substrate. Among the tested mushrooms, Ganoderma lucidium and Lentinus edodes showed the weak inhibitory activities against MAO-B.