• Title/Summary/Keyword: cell fractions

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Anti-invasive Effect of Artemisia scoparia Halophyte Extract and its Solvent-partitioned Fractions in Human Fibrosarcoma Cells (인간 섬유육종세포에서 비쑥 추출물과 유기용매 분획물의 암전이 억제 효과)

  • Kim, Junse;Kong, Chang-Suk;Sim, Hyun-Bo;Seo, Youngwan
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1100-1109
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    • 2021
  • The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anticholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H2O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.

Antimutagenic Activities of Cell Wall and Cytosol Fractions of Lactic Acid Bacteria Isolated from Kimchi

  • Park, Kun-Young;Kim, So-Hee;Son, Tae-Jin
    • Preventive Nutrition and Food Science
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    • v.3 no.4
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    • pp.329-333
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    • 1998
  • Cell wall (lactic acid bacteria-sonicated precipitate ; LAB-SP) and cytosoll(lactic acid bacteria-sonicated supernatant ; LAB-SS) fractions were prepared from kimchi fermenting lactic acid bacteria such as Leuconostoc mesenteroides, Lactobacillus brevis, Lactobacillus fermentum , Lactobacillus plantarum and Pediococcus acidilactici, with Lactobacillus acidophillus isolated from yogurt. Using the Ames mutagenicity test and SOS chormotest system, the antimutagenic acitivity of those cell fractions was studied . One hundered eighty $\mu$l of LAB-SP from lactic acid bacteria isolated from kimchi, excepting Pediococcus acidilactici, supressed the mutagenicity of 4-nitroquinoline-1-oxide(4-NQO) in Ames mutagenicity test and SOS chromotes system , by above 90% and 60% , respectively. LAB-SP from lactic acid bacteria also inhibited the mutagenicity mediated by 3-amino-1-methyl-5H-pyrido [4,3-b]indole (Trp-P-2). Lactobacillus fermentum, Lactobacillus plantarum, and Lactobacillus acidphillus had higher antimutagenicity against Trp-P-2). Lactobacillus fermentum , Lactobacillus plantarum , and Lactobacillus acidphillus had higher antimutagenicity against Trp-P-2 than the other lactic acid bacteria. However, LAB-SS of lactic acid bacteria did not show any mutagenic activity against 4-NQO in Ames mutagenicity test and SOS chromotest systems. On the mutagenicity of MEIQ and Trp-P-2 , LAB-SS of lactic acid bacteria from kimchi or dairy products exhibited a weaker inhibitory effect than LAB-SP of those bacteria. These results represent that, whether the lactic acid bacteria from kimchi are viable or nonviable, antimutagenic acitivity was still effective. We suggest that the strong, antimutaganic activity of lactic acid bacteria might be found in the cell wall fraction , rather than in the cytosol fraction.

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The Influence of Alpha-fetoprotein on Natural Suppressor Cell Activity and Ehrlich Carcinoma Growth

  • Belyaev, Nikolai Nikolaevich;Bogdanov, Andrei-Yurievich;Savvulidi, PhiIipp-Gorgievich;Krasnoshtanov, Vladimir-Konstantinovich;Tleulieva, Raikhan-Tleulievna;Alipov, Gabit-Kaimovich;Sekine, Ichiro;Bae, Jun-Sang;Lee, Jeong-Beom;Min, Young-Ki;Yang, Hun-Mo
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.4
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    • pp.193-197
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    • 2008
  • The influence of alpha-fetoprotein (AFP) on the bone marrow (BM) natural suppressor (NS) cells of intact Ehrlich carcinoma -bearing CBA mice was studied. Bone marrow NS cells were fractionated into three fractions by isopycnic centrifugation on percoll gradients: NS1 (${\rho}$=1.080 g/ml), NS2 (${\rho}$=1.090 g/ml) and NS3 (1.100> ${\rho}$ > 1.090 g/ml). These fractions were highly different in their sensitivity to known NS cell inductors (interleukin (IL)-2, IL-3 or histamine). None of the NS fractions isolated from the intact mice spontaneously produced antiproliferative activity, however, they showed a high level of NS (antiproliferative and natural killer cell inhibitory) activity under the influence of AFP. A single injection of AFP to intact mice led to an increase of spontaneous NS activity and the inhibition of natural killer cell activity. NS activity, especially NS2, was increased in when tumor cells were subcutaneously inoculated three days after AFP injection. In the AFP-treated mice, the tumor mass at 14 days was 60% larger than that in the untreated mice. Our data confirmed that AFP is a tumor marker that can inhibit cancer immunity and plays a role in cancer pathogenesis.

A Study on the Radiosensitivity and Chemosensitivity of YAC-1 Cell Line in Vitro (시험관내 YAC-1 세포주의 방사선 및 항암제감수성에 관한 연구)

  • Choi Eui-Hwan;Koh Kwang-Joon
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.43-53
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    • 1997
  • The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for mouse lymphoma YAC-1 cell line using semi automated MTT assay. 2, 4,6, 8, 10Gy were irradiated at a dose rate of 210cGy/rnin using /sup 60/Co Irradiator ALDORADO 8. After irradiation, YAC-1 cell lines(3×10⁴cells/ml) were exposed to bleomycin or cisplatin for 1 hour. The viable cells were determined for each radiation dose and/or each concentration of drug at the 4th day. And they were compared to control values. The obtained results were as follows : 1. The surviving curve with gentle slope was obtained after irradiation of 2, 4, 6, 8, 10 Gy on YAC-1 cell line. 2. The cytotoxicity of bleomycin or cisplatin was increased significantly at all concentration of 0.2㎍/ml, 2㎍/ml and 20㎍/ml on YAC-1 cell line (P<0.01). And the cytotoxicity of cisplatin was greater than that of bleomycin at all concentration on YAC-1 cell line (P<0.01). 3. There were no significant differences of surviving fractions among 4Gy, 6Gy and 8Gy after irradiation of each radiation dose with 2㎍/ml of bleomycin compared with irradiation only on YAC-1 cell line. 4. There was significant difference of surviving fraction between 2Gy and 10Gy after irradiation of each radiation dose with 2㎍/ml of cisplatin compared with irradiation only on YAC-1 cell line(P<0.05). 5. There were significant differences of surviving fractions between the groups of irradiation only and the groups of irradiation with 2㎍/ml of bleomycin or cisplatin at all doses of 2, 4, 6, 8 and 10Gy on YAC-1 cell line(P<0.05).

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Verification of Cytotoxicity Against Cancer Cell Line and Estrogen-like Activity of Cheongkukjang (청국장의 암세포주에 대한 세포독성 및 에스트로겐 유사활성 검증)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.153-157
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    • 2007
  • In order to evaluate the cancer preventive and estrogenic compounds in soybean and Cheongkukjang, MTT assay and in vitro test system for the evaluation of the estrogenic activity were applied. The fractions from the ethanol extract of soybean and Cheongkukjang were prepared by the systematic extraction procedure with the solvents such as hexane, ethyl ether, butanol, methanol and H$_2$O. Ethyl ether fractions of soybean and Cheongkukjang showed the highest cytotoxicity against U937 cell line in dose dependent manner, and ethyl ether fraction of Cheongkukjang showed two times higher cytotoxicity than that of soybean. Aqueous fraction of soybean and ethyl ether fraction of Cheongkukjang revealed the highest estrogenic activity and activity was higher in the fractions of Cheongkukjang than soybean. Mixture of Spirulina and Cheongkukjang showed synergistic activity. These observations concerning cancer preventive and estrogen effects of soybean and Cheongkukjang suggest that these materials possess useful ingredients for the prevention of cancer and/or postmenopausal disorder.

Antimutagenic and Antileukemic Activities Aloe vera L.

  • Lee, Keyong-Ho;Kang, Hee-Gon;Cho, Choa-Hyoung;Lee, Moon-Joon;Lee, Jae-Hyun;Kim, Chang-Han
    • Natural Product Sciences
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    • v.6 no.2
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    • pp.56-60
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    • 2000
  • We investigated that the extract of Aloe vera L. and its fractions exert antimutagenic activity against Salmonella typhimurium TA98 and TA100, and antileukemic effect against K562 human leukemia cell line. The aqueous ethanolic extract of A. vera L. was revealed to have antimutagenic effect on the AF-2 (2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide) in Salmonella mutation assay. Among the three fractions (fractions A, B and C) separated by silica gel chromatography, fraction C $(50\;{\mu}g/plate)$ exhibited the greatest antimutagenic effect on the AF-2 with inhibition rate of 84 and 90% in Salmonella typhimurium TA98 and TA100, respectively. The fraction C $(500\;{\mu}g/ml)$ inhibited the growth of K562 human leukemia cell line by 93% in MTT assay. However, the components of A. vera L. did not exhibit cytotoxic effect against MDBK bovine normal kidney in MTT assay.

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Antibacterial Activity and Nitric Oxide Production Inhibitory Activity of the Extract and its Fractions from the Leaves of Prunus sargentii (산벚나무 잎 추출물 및 분획물의 항균활성과 Nitric Oxide 생성억제 활성)

  • Yang, Sun-A;Pyo, Byoung-Sik;Kim, Sun-Min;Lee, Kyoung-In
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.5
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    • pp.308-314
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    • 2012
  • This study was carried out to investigate the antibacterial activity against pathogens of acne and the anti-inflammatory effect of 75% ethanol extract and its fractions from the leaves of Prunus sargentii. In the antibacterial activity by the disc diffusion assay, the extract showed the highest effect against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis in 5 mg/disc. However, the ethyl acetate fraction showed the highest antibacterial activity in 1 mg/disc. On the other hand, the hexane and chloroform fraction showed strong nitric oxide (NO) production inhibitory effect in lipopolysaccharide (LPS)-stimulated Raw 264.7 cell. In the cell viability of Raw 264.7 by MTT assay, the extract and all fractions were exhibited normal viabilities as nontoxic result. Consequently, the extract from the leaves of P. sargentii and its ethyl acetate fraction could be applicable to functional materials for antibacterial activity related fields. Moreover, the hexane and chloroform fraction could be applicable to candidate materials as anti-inflammatory agent.

Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

  • Farzaneh, Mohsen;Shi, Zhi-Qi;Ahmadzadeh, Masoud;Hu, Liang-Bin;Ghassempour, Alireza
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.209-215
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    • 2016
  • In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1), caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC) method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut.

Cytoprotective Effects and Gene Expression Patterns Observed Based on the Antioxidant Activity of Lonicera japonica Extract (금은화 추출물의 항산화 효과를 통한 세포 보호효과 및 유전자 발현 양상)

  • Cho, Won June;Yoon, Hee Seung;Kim, Yong Hyun;Kim, Jung Min;Yoo, Il Jae;Han, Man-Deuk;Bang, In Seok
    • Journal of Life Science
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    • v.23 no.8
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    • pp.989-997
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    • 2013
  • In this study, based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of L. japonica, the protective cellular effects and gene expression patterns of ethyl acetate fractions on $H_2O_2$-induced Raw 264.7 cell death ($IC_{50}$) were analyzed. The antioxidant activity of the fractions measured using DPPH free radical scavenging activity increased in a dose-dependent manner, and the $ED_{50}$ exhibited the highest $39.56{\mu}g/ml$ in the ethyl acetate fraction. In addition, the ethyl acetate fractions' cell viability on $H_2O_2$-induced Raw 264.7 cell damage increased in a concentration-dependent manner, showed a visible cell survival rate of 82.49% at a concentration of $100{\mu}g/ml$. The gene expression patterns related to the ethyl acetate fractions' cytoprotective effect in $H_2O_2$-induced Raw 264.7 cell damage presented similar patterns to those of BHA. In comparative analysis for antioxidant activity-related genes affected by ethyl acetate fractions and BHA in $H_2O_2$-induced Raw 264.7 cells, both ethyl acetate fractions and BHA showed very similar gene expression patterns, but the gene expression level of the heme oxygenase 1 (Hmox1) gene making antioxidant enzymes in cells was four times higher in ethyl acetate fractions than BHA. In inflammation-related genes in $H_2O_2$induced Raw 264.7 cells, the T-box transcription factor (Tbx21) gene was expressed about two times more frequently in the ethyl acetate fraction treatment group, while it was expressed half as frequently in the BHA treatment group.

Antioxidant and Neuroprotective Effects of Perilla frutescens var. japonica Leaves (들깨 잎 추출물의 항산화 및 신경세포 보호작용)

  • Lee, Jong-Im;Jin, Chang-Bae;Ryu, Jae-Ha;Cho, Jung-Sook
    • YAKHAK HOEJI
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    • v.52 no.2
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    • pp.117-124
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    • 2008
  • The leaves of Perilla frutescens Britt. var. japonica Hara (Labiatae) are often used in gourmet food in several Asian countries. Two kinds of perilla cultivars, Namcheon (NC) and Bora (BR), have been respectively developed in Korea by the pure line of 'deulkkae' from the local variety and by the cross of 'deulkkae' and 'chajogi'. The present study evaluated and compared antioxidant and neuroprotective effects of the fractions prepared from the leaves of the two cultivars using cell-free bioassay systems and primary cultured rat cortical cells. We found that the spirit, chloroform, hexane and butanol fractions from NC and BR leaves inhibited lipid peroxidation initiated in rat brain homogenates by $Fe^{2+}$ and L-ascorbic acid. In contrast, only the spirit and butanol fractions from both cultivars exhibited 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Among the fractions tested, the butanol fractions from NC and BR leaves exhibited the most potent antioxidant properties, and the butanol fraction from BR was more potent than the NC fraction. In consistence with these findings, the butanol fractions from both cultivars protected primary cultured cortical cells from the oxidative damage induced by $H_2O_2$ or xanthine and xanthine oxidase, with the BR butanol fraction being more active. The butanol fractions from NC and BR did not produce cytotoxicity in our cultures treated for 24 h at the concentrations of up to $100\;{\mu}g/ml$. Taken together, these results indicate that the leaves of the two cultivars of Perilla frutescens exert antioxidant and neuroprotective effects, and that the butanol fraction from BR leaves exhibits the most potent antioxidative neuroprotection among the fractions tested in this study.