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Inhibitory effects of Zanthoxylum piperitum on the LPS-induced production of nitric oxide and proinflammatory cytokines in RAW264.7 cells (초피(椒皮)의 RAW264.7세포에서의 LPS에 의해 유도되는 nitric oxide 및 전염증사이토카인 생성억제효과)

  • Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.69-76
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    • 2006
  • Objectives : The fresh young leaves and dried fruits of Zanthoxylum piperitum (Korean name: Chopi) are used as diuretics, stomachies, anthelmintic and for the treatments of disorders of the digestive organ in Asia. We investigated inhibitory effects of Zanthoxylum piperitum extract on lipopolysaccharide(LPS)-induced production of nitric oxide(NO) and pro-inflammatory cytokines including $TNF-{\alpha}$ and $IL-1{\beta}$ from RAW264.7 mouse macrophage cells. Methods : After methanol extract of Zanthoxylum Fructus (Zanthoxylum extract) was pretreated in RAW264.7 cells, the cells were stimulated with LPS. Cell toxicity of Zanthoxylum extract was assayed bv MTT assay. The production of NO from the cells was measured in culture medium by Griess reaction. The production of $TNF-{\alpha}$ and $IL-1 \;{\beta}$ from the cells was measured in culture medium by ELISA. Results : Zanthoxylum Fructus extract greatly inhibited the production of inflammatory mediators such as NO, $TNF-{\alpha}$ and $IL-1{\beta}$ from LPS-stimulated RAW264.7 cells. Conclusion : This result suggests that Zanthoxylum extract may have an anti-inflammatory effect through the inhibition of inflammatory mediators.

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Isolation of Xanthine Oxidase Inhibitors from Ginkgo biloba Leaves-Derived Components

  • Baek, Bong-Rae;Kim, Moo-Key;Lee, Sung-Eun;Hwang, Young-Hee;Lee, Hoi-Seon
    • Preventive Nutrition and Food Science
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    • v.7 no.1
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    • pp.18-21
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    • 2002
  • The extract of Ginkgo biloba leaves was measured for inhibitory activity against xanthine oxidase. Aceton extract of G. biloba leaves showed strong inhibitory activity. Inhibitory activities of the fractionated extract were in the order of water > ethyl acetate fractions. Two fractions exhibiting strong inhibitory activities ware further purified via repeated silica gel, Amberlite IRN-78, Polyclar AT, and Sephadex LH-20 column chromatographies. Active components were isolated and identified through $^1$H-NMR and $^{13}$ C-NMR. The compounds were characterized as kaempferol 3-Ο-$\alpha$-(6$^{"′}$-p-coumaroylglucosyl-$\beta$-1,4-rhamnoside), and quercertin 3-Ο-$\alpha$-(6$^{′}$-p-coumaroylglucosyl-$\beta$-1,4-rhamnoside).

Interactions between Water-Soluble Polyparacyclophanes and Drugs (III) -Complex Formation of Water-Soluble Polyparacyclophanes with Fluorescent Hydrophobic Naphthalene Derivatives in Aqueous Solution- (수용성 폴리파라시클로판류와 약물과의 상호작용(제 3보)-수용액 중 수용성 폴리파라시클로판류와 형광 소수 나프탈렌 유도체류와의 복합체 형성-)

  • Chun, In-Koo;Lee, Min-Hwa
    • Journal of Pharmaceutical Investigation
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    • v.19 no.2
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    • pp.71-79
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    • 1989
  • Complex formation of water-soluble polyparacyclophanes bearing two diphenylmethane or two diphenyl ether skeletons with l-anilinonaphthalene-8-sulfonate (ANS) and 2-p-toluidinylnaphthalene-6-sulfonate (TNS) was investigated quantitatively to develop useful host compounds comparing with ${\alpha}\;-\;and\;{\beta}-cyc1odextrins$$({\alpha}-\;and\;{\beta}-CyDs$) in aqueous solution. Benesi-Hildebrand type analysis of the fluorescent intensity showed that the dissociation constants (Kd) of paracyclophane-ANS complexes were $1.55\;{\times}\;10^{-4}M$ for 1,6,20,25-tetraaza[6.1.6.1]paracyclophane(CPM 44) and $1.23\;{\times}\;10^{-4}M$ for 1,7,21,27-tetraaza[7.1.7.1]paracyclophane (CPM 55), and those of paracyclophane-TNS complexes were $6.99\;{\times}\;10^{-6}M$ for CPM 44 and $6.23\;{\times}\;10^{-5}M$ for CPM 55, in 1:1 molar ratio. On the other hand, the Kd values of 1,7,21,27-tetraaza-14,34-dioxa[7.1.7.1]paracyclophane (CPE 55)-ANS, 1,8,22,29-tetraaza-15,36-dioxa[8.1.8.1]paracyclophane (CPE 66)-ANS, CPE 55-TNS, CPE 66-TNS complexes were $1.75\;{\times}\;10^{-3}M$, $3.07\;{\times}\;10^{-3}M$, $3.75\;{\times}\;10^{-3}M$ and $2.15\;{\times}\;10^{-3}M$, respectively. On the contrary, the Kd values of ${\alpha}-CyD-ANS$, ${\beta}-CyD-ANS$, ${\alpha}-CyD-TNS$ and ${\beta}-CyD-TNS$ complexes were found to be $3.98\;{\times}\;10^{-2}M$, $1.05\;{\times}\;10^{-2}M$, $1.38\;{\times}\;10^{-2}M$ and $3.52\;{\times}\;10^{-4}M$, respectively. These results mean that the complexation of CPMs with ANS or TNS is by 5.6-1,975 fold stronger than that for ${\alpha}-or\;{\beta}-CyDs$, and the complex formation of CPEs with ANS or TNS is nearly same as or somewhat stronger than that for ${\alpha}-or\;{\beta}-CyDs$. From the Kd values determined at different temperatures, thermodynamic parameters were calculated and the complexation was found to be a spontaneous exothermic reaction. The effects of pH on Kd values of CPM 44-ANS, and CPM 55-ANS complexes were negligible in the range of pH 1.2-1.8. However, the Kd values of these complexes increased significantly with increasing ionic strength.

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Anticomplement Activities of Oleanolic Acid Monodesmosides and Bisdesmosides Isolated from Tiarella polyphylla

  • Park, Si-Hyung;Oh, Sei-Ryang;Jung, Keun-Young;Lee, Im-Seon;Ahn, Kyung-Seop;Kim, Jae-Gil;Lee, Jung-Joon;Lee, Hyeong-Kyu
    • Archives of Pharmacal Research
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    • v.22 no.4
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    • pp.428-431
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    • 1999
  • Seven known oleanolic acid glycosides (1-7) were isolated form the MeOH extract of Tiarella polyphylla. The structures were identified to be 3-O-($\beta$-glucopyranosyl) oleanolic acid (1), 3-O-[$\beta$-D-glucopyranosyl-(1 3)-$\beta$-D-glucopyranosyl] oleanolic acid (2), 3-O-D-[$\beta$-D-glucopyranosyl-(1 2)-$\beta$-D-glycopyranosyl] oleanolic acid (3), 3-O-[$\beta$-D-glucopyranosyl-(1 3)-$\beta$-D-glucopyranosyl] oleanolic acid 28-O-$\beta$D-glucopyranosyl ester (4), 3-O-[$\beta$-D-glucopyranosyl-(1 2)-$\beta$-D-glucopyranosyl] oleanolic acid 28-O-$\beta$-D-glucopyranosyl ester (5), 3-O-[a-L-rahmnopyranosyl-(1 3)-$\beta$-D-glucururonopyranosyl] oleanolic acid (6), and 3-O-[$\alpha$-L-rhamnopyranosyl-(1 3)-$\alpha$-D-glucuronopyranosyl] oleanolic acid 28-O-$\alpha$-D-glucopyranosyl ester (7) on the basis of physicochemical and spectral data. These triterpene glycosides were tested for the anti-complement activity and hemolytic activity. Bisdesmosidic saponins, 4, 5, and 7, showed anti-complement activity; in contrast, monodesmosidic saponins, 1-3, and 6, showed direct hemolytic activity. Methyl esterified monodesmosidic saponins showed anti-complement activity at a low concentration and hemolytic activity at a high concentration.

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Quantitative Changes of Cholesterol Oxides Formed in Butter under Varied Storage Conditions (상이한 조건에서 저장한 버터로부터 생성된 콜레스테롤 산화물의 양적변화)

  • Chang, Young-Sang;Yang, Joo-Hong;Shin, Hyo-Sun
    • Korean Journal of Food Science and Technology
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    • v.22 no.7
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    • pp.767-773
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    • 1990
  • The effect of storage conditions on the oxidative stability of cholesterol in butter was studied by quantifying cholesterol oxides by GC-MS. Experimental variables for storage conditions were packaging, storage temperature, light source, and storage period. No cholesterol oxides were detected from packaged butter under all storage conditions. When unpackaged butter was stored under ultraviolet light at ambient temperature, $7{\alpha}-hydroxycholesterol,\;7{\beta}-hydroxycholesterol$, cholesterol ${\beta}-epoxide$, cholesterol ${\alpha}-epoxide$, cholestane-triol were detected after 2, 4, 6 and 8 weeks of storage. The amounts of cholesterol oxide species produced were different depending on the storage periods. The amounts of each cholesterol oxides, $7{\alpha}-hydroxycholesterol,\;7{\beta}-hydroxycholesterol$, cholesterol ${\beta}-epoxide$, cholesterol ${\alpha}-epoxide$, and cholestane-triol, produced after 2 weeks of storage were 34.2, 14.0, 12.1, 1.30, and 0.50 ppm, respectively, and after 8 weeks of storage were 68.1, 29.1, 56,3, 8.50, and 4.00 ppm, respectively with trace amounts of 3,5-cholestadien-7-one. When fluorescent light was used instead of ultraviolet light with other conditions remained the same, the same species of cholesterol oxides were detected but with lesser amounts.

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Phytochemical Studies on Astragalus Root (3);Triterpenoids and Sterols (황기의 성분연구 (3);Triterpenoids and Sterols)

  • Jung, Hye-Sil;Lee, Eun-Ju;Lee, Je-Hyun;Kim, Ju-Sun;Kang, Sam-Sik
    • Korean Journal of Pharmacognosy
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    • v.39 no.3
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    • pp.186-193
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    • 2008
  • Astragali Radix, known as Huangqi, is the most important tonic in the traditional oriental medicine. It reinforces 'qi' (vital energy), strengthens the superficial resistance and promotes the discharge of pus and the growth of new tissue. It has long been used as an anti-perspirant, anti-diuretic or a tonic. Eleven compounds were isolated from the hexane and EtOAc fractions from the roots of Astragalus membranaceus (Leguminosae) and their structures were identified as four triterpenoids [lupenone (1), friedelin (2), lupeol (3), soyasapogenol E (9)] and seven sterols [${\beta}-sitosterol$ (4), stigmastane-3,6-dione (5), $7{\alpha}-hydroxysitosterol$ (6), $5{\alpha},6{\beta}-dihydroxysitosterol$ (7), $7-oxo-{\beta}-sitosterol$ (8), ${\beta}-sitosterol$ glucoside 6'-O-palmitate (10), ${\beta}-sitosterol$ glucoside (11)]. The chemical structures of these compounds were identified on the basis of spectroscopic methods and comparison with literature values. Among these compounds, lupenone (1), friedelin (2), lupeol (3), stigmastane-3,6-dione (5), $7{\alpha}-hydroxysitosterol$ (6), $5{\alpha},6{\beta}-dihydroxysitosterol$ (7), $7-oxo-{\beta}-sitosterol$ (8), soyasapogenol E (9), and ${\beta}-sitosterol$ glucoside 6'-O-palmitate (10) were isolated from this plant for the first time.

Silica induced Expression of IL-1$\beta$, IL-6, TNF-$\beta$, TGF-$\alpha$, in the Experimental Murine Lung Fibrosis (유리규산에 의한 폐장내 IL-1$\beta$, IL-6, TNF-$\alpha$, TGF-$\beta$의 발현)

  • Ki, Shin-Young;Park, Sung-Woo;Lee, Myung-Ran;Kim, Eun-Young;Uh, Soo-Taek;Kim, Yong-Hoon;Park, Choon-Sik;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.835-845
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    • 1998
  • Background: Silica-induced lung diseases is characterized by the accumulation of inflammatory cells at early stage and fibrosis in pulmonary parenchyma and interstitium at late stage. As a consequence of inflammation, silicosis is accompanied with the expansion of interstitial collagen and the formation of fibrotic nodule. In this process, several kinds of lung cells produce cytokines which can amplify and modulate pulmonary fibrosis. The alveolar macrophage is a potent source of proflammatory cytokines and growth factor. But in the process of silicotic inflammation and fibrosis, there are many changes of the kinetics in cytokine network. And the sources of cytokines in each phase are not well known. Method: 2.5 mg of silica was instillated into the lung of C57BL/6J mice. After intratracheal instillation of silica, the lungs were removed for imunohistochemical stain at 1, 2, 7 day, 2, 4, 8, 12 week, respectively. We investigated the expression of IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ in lung tissue. Results: 1) The expression of IL-6 increased from 1 day after exposure to 8 weeks in vascular endothelium. Also peribronchial area were stained for IL-6 from 7 days and reached the peak level for 4 weeks. 2) The IL-1 $\beta$ was expressed weakly at the alveolar and peribronchial area through 12 weeks. 3) The TNF-$\alpha$ expressed strongly at alveolar and bronchial epithelia during early stage and maintained for 12 weeks. 4) TGF-$\beta$ was expressed strongly at bronchial epithelia and peribronchial area after 1 week and the strongest at 8 weeks. Conclusion: The results above suggests IL-6, TNF-$\alpha$ appear to be a early inflammatory response in silica induced lung fibrosis and TGF-$\beta$ play a major role in the maintenance and modulation of fibrosis in lung tissue. And the regulation of TNF-$\alpha$ production will be a key role in modultion of silica-induced fibrosis.

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Structural Characterization of a Flavonoid Compound Scavenging Superoxide Anion Radical Isolated from Capsella bursa-pastoris

  • Kweon, Mee-Hyang;Kwak, Jae-Hyock;Ra, Kyung-Soo;Sung, Ha-Chin;Yang, Han-Chul
    • BMB Reports
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    • v.29 no.5
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    • pp.423-428
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    • 1996
  • A superoxide anion radical scavenger isolated from Capsella bursa-pastoris was characterized by infrared (IR) spectroscopy, sugar analysis, ultraviolet (UV) spectroscopy, $^{1}H$ and $^{13}C$ nuclear magnetic resonance (NMR) spectroscopies, and fast atom bombardment (FAB) mass analysis. The compound was assumed to be a flavonoid-O-glycoside from IR spectrum and UV absorption maxima. When the sugar composition of the compound was examined by thin layer chromatography (TLC) and gas chromatography (GC) of the acid hydrolysate, only glucose was detected. According to the results of UV spectrotroscopy by using shift reagents, the compound was supposed to be luteolin (5,7,3',4'-tetrahydroxy flavone) or chrysoeriol (5,7,4'-trihydroxy-3'-methoxy flavone) with glucose. Based on $^{1}H$- and $^{13}C-NMR$ spectroscopies, the compound was deduced as 7,4'-dihydroxy-5,3'-dimethoxy-${\alpha}$-6-c-glucosyl-${\beta}$-2"-o-glucosyl flavone. In FAB mass analysis the compound was finally characterized as 7,4'-dihydroxy-5,3'-dimethoxy-${\alpha}$-6-c-glucosyl-${\beta}$-2"-o-glucosyl flavone ($C_{29}H_{34}O_{16}$, M.W.=638).

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Effect of Egg White Combined with Chalcanthite on Lipopolysaccharide induced Inflammatory Cytokine Expression in RAW 264.7 cells

  • Choi, Eun-A;Yoon, Jeung-Won;Choi, Hak-Joo;Kim, Dong-Hee;Yoo, Hwa-Seung
    • Journal of Pharmacopuncture
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    • v.15 no.1
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    • pp.7-11
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    • 2012
  • Aim: Historically, mineral compound herbal medicines have long been used in treatments of immune-related diseases in Korea, China and other Asian countries. In this study, we investigated the anti-inflammatory effect of egg white combined with chalcanthite (IS4) on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: RAW 264.7 cells cultured with LPS and various concentrations of IS4 were analyzed to determine the production of pro-inflammatory cytokines and mediators by using enzyme-linked immune sorbent assays (ELISAs). Results: IS4 concentration inhibited the production of interleukin-1beta (IL-$1{\beta}$), interleukin-6 (IL-6), and granulocyte-macrophage colony-stimulating factor (GM-CSF) induced by LPS. IS4 at high concentrations (25 and 50 ${\mu}g/ml$) inhibited, in concentration-dependent manner, the expression of tumor necrosis factor-alpha (TNF-${\alpha}$) stimulated by LPS. Conclusion: IS4 has shown an anti-inflammatory effect in RAW 264.7 cells.

Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.