• Title/Summary/Keyword: HF assay

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Anti-cancer Activities of Extract from the Bark of Melia azedarach L. var. japonica Makino (고련피 추출물의 항암활성)

  • Kim, Hyun-Woo;Kang, Se-Chan
    • Korean Journal of Plant Resources
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    • v.22 no.4
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    • pp.312-316
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    • 2009
  • In the present study, the anti-cancer activity of 80% ethanol extracts from 120 kinds of medicinal herbs and native plants were investigated. Among them, the barks of Melia azedarach L. var. japonica Makino showed the highest cytotoxicity in HCT-15 human colon cancer cell. With this result, we carried out hollow fiber (HF) assay and anti-metastasis study to confirm the anti-cancer effects of M. azedarach var. japonica. In MTT assay, M. azedarach var. japonica.inhibited the proliferation of HCT-15 cells in dose-dependent manner. HF assay was carried out using A549 human adenocarcinoma cell, HCT-15 and SK-Hep1 human liver cancer cell via intraperitoneal (IP) and subcutaneous (SC) site. As a results, SK-Hep1 implanted in IP site showed the highest cytotoxicity. The result from metastatic model using B16/BL6 mouse corresponded to that of HF assay. These results suggest that the ethanol extract from M. azedarach var. japonica. might have a potent anti-cancer activity and advanced study is needed for the development of novel natural anti-cancer drug.

A Study on the Cytotoxic Effects of Several Plant Extracts on the Cell viability and Cell Adhesion Activity in Cultured NIH3T3 Fibroblast (몇 가지 식물추출물이 배양 NIH3T3 섬유모세포의 세포생존율과 세포부착률에 미치는 세포독성에 관한 연구)

  • Rim, Yo-Sup;Song, Won-Seob;Seo, Young-Mi;Park, Seung-Taeck;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.3
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    • pp.116-124
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    • 2010
  • This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

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Evaluation of Genotoxicity of Three Antimalarial Drugs Amodiaquine, Mefloquine and Halofantrine in Rat Liver Cells

  • Farombi E. Olatunde
    • Environmental Mutagens and Carcinogens
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    • v.25 no.3
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    • pp.97-103
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    • 2005
  • The genotoxic effect of antimalarial drugs amodiaquine (AQ), mefloquine (MQ) and halofantrine (HF) was investigated in.at liver cells using the alkaline comet assay. AQ, MQ and HF at concentrations between $0-1000{\mu}mol/L$ significantly increased DNA strand breaks of rat liver cells dose-dependently. The order of induction of strand breaks was AQ>MQ>HF. The rat liver cells exposed to AQ and HF (200 and 400 ${\mu}mol/L$) and treated with (Fpg) the bacterial DNA repair enzyme that recognizes oxidized purine showed greater DNA damage than those not treated with the enzyme, providing evidence that AQ and HF induced oxidation of purines. Such an effect was not observed when MQ was treated with the enzyme. Treatment of cells with catalase, an enzyme inactivating hydrogen peroxide, decreased significantly the extent of DNA damage induced by AQ, and HF but not the one induced by MQ. Similarly quercetin, an antioxidant flavonoid at $50{\mu}mol/L$ attenuated the extent of the formation of DNA strand breaks by both AQ and HE. Quercetin, however, did not modify the effects of MQ. These results indicate the genotoxicity of AQ, MQ and HF in rat liver cells. In addition, the results suggest that reactive oxygen species may be involved in the formation of DNA lesions induced by AQ and HF and that, free radical scavengers may elicit protective effects against genotoxicity of these antimalarial drugs.

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Benzoylaconine improves mitochondrial function in oxygen-glucose deprivation and reperfusion-induced cardiomyocyte injury by activation of the AMPK/PGC-1 axis

  • Chen, Leijie;Yan, Laixing;Zhang, Weiwei
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.5
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    • pp.325-333
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    • 2022
  • Heart failure (HF) has become one of the severe public health problems. The detailed role of mitochondrial function in HF was still unclear. Benzoylaconine (BAC) is a traditional Chinese medicine, but its role in HF still needs to be explored. In this study, oxygen-glucose deprivation and reperfusion (OGD/R) was executed to mimic the injury of H9C2 cells in HF. The viability of H9C2 cells was assessed via MTT assay. OGD/R treatment markedly decreased the viability of H9C2 cells, but BAC treatment evidently increased the viability of OGD/R-treated H9C2 cells. The apoptosis of H9C2 was enhanced by OGD/R treatment but suppressed by BAC treatment. The mitochondrial membrane potential was evaluated via JC-1 assay. BAC improved the mitochondrial function and suppressed oxidative stress in OGD/R-treated H9C2 cells. Moreover, Western blot analysis revealed that the protein expression of p-AMPK and PGC-1α were reduced in OGD/R-treated H9C2 cells, which was reversed by BAC. Rescue assays indicated that AMPK attenuation reversed the BAC-mediated protective effect on OGD/R-treated cardiomyocytes. Moreover, BAC alleviated myocardial injury in vivo. In a word, BAC modulated the mitochondrial function in OGD/R-induced cardiomyocyte injury by activation of the AMPK/PGC-1 axis. The findings might provide support for the application of BAC in the treatment of HF.

The Toxicity and Anti-cancer Activity of the Hexane Layer of Melia azedarach L. var. japonica Makino's Bark Extract

  • Kim, Hyun-Woo;Kang, Se-Chan
    • Toxicological Research
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    • v.28 no.1
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    • pp.57-65
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    • 2012
  • In this study, the 4-week oral toxicity and anti-cancer activity of the hexane layer of Melia azedarach L. var. japonica Makino's bark extract were investigated. We carried out a hollow fiber (HF) assay and 28-day repeated toxicity study to confirm the anti-cancer effect and safety of the hexane layer. The HF assay was carried out using an A549 human adenocarcinoma cell via intraperitoneal (IP) site with or without cisplatin. In the result, the 200 mg/kg b.w of hexane layer with 4 mg/kg b.w of cisplatin treated group, showed the highest cytotoxicity aginst A549 carcinoma cells. For the 28-day repeated toxicity study, 6 groups of 10 male and female mice were given by gavage 200, 100, or 50 mg/kg b.w hexane layer with or without 4 mg/kg b.w of cisplatin against body weight, and were then sacrificed for blood and tissue sampling. The subacute oral toxicity study in mice with doses of 200, 100, and 50 mg/kg b.w hexane layer showed no significant changes in body weight gain and general behavior. The cisplatin-treated group significantly decreased in body weight compared to the control group but regained weight with 100 and 200 mg/kg b.w of hexane layer. The biochemical analysis showed significant increase in several parameters (ALT, total billirubin, AST, creatinine, and BUN) in cisplatin-treated groups. However, in the group given a co-treatment of hexane layer (200 mg/kg b.w), levels of these parameters decreased. In hematological analysis, cisplatin induced the reduction of WBCs and neutrophils but co-treatment with hexane layer (100 and 200 mg/kg b.w) improved these toxicities caused by cisplatin. The histological profile of the livers showed eosinophilic cell foci in central vein and portal triad in cisplatin treated mice. These results show that hexane layer might have an anti-cancer activity and could improve the toxicity of cisplatin.

The Effects on Antimicrobial and Cytotoxicity of Hijikia Fusiformis Fraction (톳분획물의 항균 및 암세포 증식 억제효과)

  • Shon, Jae-Hak;Kang, Dae-Yeon;Oh, Hyun-Cheol;Jung, Bok-Mi;Kim, Mi-Hyang;Shin, Mi-Ok;Bae, Song-Ja
    • Journal of Nutrition and Health
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    • v.39 no.5
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    • pp.444-450
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    • 2006
  • In this study, we investigated antimicrobial and cytotoxicity effects to each· fraction extracted from Hizikiafusifonnis (HF), which were extracted methanol (HFM) and then the extract was fractionated into four different types: hexane (HFMH) , methanol (HFMM) , bulanol (HFMB) and aquous (HFMA) partition layers. We determined the cytotoxic effect of these layers on human cancer cells by MIT assay. Among various partition layers of HF, the HFMB and HFMM were showed the strong cytotoxic effects on cancer cell lines we used. The quinone reductase (QR) induced activity of the HFMB on HcpG2 cells at $150\;{\mu}g/mL$ concentration was 2.63 times more effective compared to the control value of 1.0. Although further studies are needed, the present work suggests that HF maybe a chemopreventive agent for the treatment of human cancer cells.

Effect of Hibisci Flos on Inflammatory Cytokines Production in lipopolysaccaride-stimulated Raw 264.7 Macrophages (목근화(木槿花) 물추출물의 항염효능에 관한 연구)

  • Lee, Dong-Min;Han, Hyo-Sang;Lee, Young-Jong
    • The Korea Journal of Herbology
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    • v.28 no.5
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    • pp.61-68
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    • 2013
  • Objectives : Hibisci Flos has long been used for inflammatory diseases in traditional Korean medicine. However, little scientific investigation has been carried out. The aim of the present study is to investigate the effect of Hibisci Flos water extract (HF) on inflammatory cytokines production in Raw 264.7 cells stimulated by lipopolysaccaride (LPS). Method : HF was prepared by extracting with boiling water for 2 hours. We observed the cell viability of mouse macrophage Raw 264.7, the production of nitric oxide (NO) and the inflammatory cytokines such as interleukin (IL)-4, IL-5, IL-10, IL-15, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interferon-gamma (IFN-${\gamma}$), vascular endothelial growth factor (VEGF), granulocyte macrophage-colony stimulating factor (GM-CSF), and macrophage colony-stimulating factor (M-CSF) in Raw 264.7 cells stimulated by LPS. Result : The MTT assay was carried out to check the cellular toxicity of HF. No significant toxicity was observed in the experiment. HF significantly inhibited the increase of NO in the macrophages induced by LPS after 24 hour treatment. HF significantly inhibited the production of IL-4, IL-5, IL-10, IL-15, TNF-${\alpha}$, IFN-${\gamma}$, VEGF, GM-CSF and M-CSF in the Raw 264.7 cells induced by LPS in the concentration of $25{\mu}g/mL$ or higher. Conclusion : These results suggest that HF might have regulatory effects on LPS-induced inflammatory cytokine production, which might explain its beneficial effect in the treatment of inflammatory disease.

Antibacterial and Antibiotic Activity Enhancing Effect of Extract and Fractions from the Root of Rumex japonicus Houtt (참소리쟁이 뿌리 추출물 및 분획의 항균 활성과 항생제 증강 활성)

  • Yang, Sun A;Kim, A Young;Pyo, Byoung Sik;Kim, Sun Min
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.6
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    • pp.375-380
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    • 2017
  • Background: The objective of this study were to determine the antibacterial activity and antibiotic activity-enhancing effect of 70% ethanol extract of the root of Rumex japonicus Houtt. and its fractions when used in combination with gentamicin against aerobic skin flora. Methods and Results: The antibacterial activity and antibiotic (gentamicin) activity enhancing effect against aerobic skin flora were determined using the disc diffusion assay. Chloroform fraction (CF) and ethyl acetate fraction (EF) showed higher activities against Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. Regarding the antibiotic (gentamicin) activity-enhancing effect against aerobic skin flora, the n-hexane fraction (HF) and CF showed strong activity. The combination of HF and CF with gentamicin was evaluated using the broth dilution assay to determine the inhibitory effect on the growth of aerobic skin flora. The combination of CF with gentamicin exhibited the highest inhibitory effect on the growth of S. aureus and S. epdermidis. MTT assay performed to determine the viability of L929 cells revealed that EF treatment resulted in viability of 33.96 - 116.76% at the tested concentration. The combination of 70% ethanol extract and its other fractions with gentamicin showed low cell toxicity. Conclusions: Appropriate use of antimicrobial agents is important prior to the development of new antibiotics. The 70% ethanol extract of the root of R. japonicus Houtt. and its fractions showed significant synergism with gentamicin when used in combination against S. aureus and S. epdermidis. Thus, R. japonicus Houtt. could be used as a functional materials in antimicrobial-related fields.

A Study on the Decomposition of Amygdalin Using an In Vitro Assay (Amygdalin의 in Vitro 분해에 관한 연구)

  • Kwon, Hoon-Jeong;Jo, Yong-Jin
    • Toxicological Research
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    • v.23 no.1
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    • pp.47-53
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    • 2007
  • Amygdalin is a cyanogenic glycoside which is commonly found in almonds, bamboo shoots, and apri-cot kernels, and peach kernels. Amygdalin was first hydrolysed into prunasin, then degraded into cyanohydrin by sequential two-stage mechanism. The objective of this study was to examine the amygdalin decomposition and cyanide formation at various in vitro conditions, including acid, enzyme and anaerobic microbes (AM) in human feces (HF). In acid hydrolysis mimicking gastric environment, amygdalin was degraded to cyanide up to 0.2% in specific pH. In contrast, enzyme assay showed higher cyanide generation either by ${\beta}$-glucosidase, or by incubation with microbe. In conclusion, we are convinced of cyanide generation are occurred mainly by microbiological activities of the gut flora up to 41.53%. After ingestion with some staff, the degree and site of degradation in an organism is a key parst of regulatory decision making of that staff.

Inhibitory effect of water-soluble mulberry leaf extract on hepatic lipid accumulation in high-fat diet-fed rats via modulation of hepatic microRNA-221/222 expression and inflammation (고지방식이 급여 쥐에서 수용성 뽕나무 잎 추출물의 간 microRNA-221/222 발현 및 염증 조절을 통한 간 지질 축적억제 효과)

  • Lee, Mak-Soon;Kim, Cheamin;Ko, Hyunmi;Kim, Yangha
    • Journal of Nutrition and Health
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    • v.55 no.2
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    • pp.227-239
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    • 2022
  • Purpose: This study investigated the effects of water-soluble mulberry leaf extract (ME) on hepatic lipid accumulation in high-fat diet-fed rats via the regulation of hepatic microRNA (miR)-221/222 and inflammation. Methods: Male Sprague-Dawley rats (4 weeks old) were randomly divided into 3 groups (n = 7 each) and fed with 10 kcal% low-fat diet (LF), 45 kcal% high-fat diet (HF), or HF + 0.8% ME for 14 weeks. Lipid profiles and cytokine levels of the liver and serum were measured using commercial enzymatic colorimetric and enzyme-linked immunosorbent assay, respectively. The messenger RNA (mRNA) and miR levels in liver tissue were assayed by real-time quantitative reverse-transcription polymerase chain reaction. Results: Supplementation of ME reduces body weight and improves the liver and serum lipid profiles as compared to the HF group. The mRNA levels of hepatic peroxisome proliferator-activated receptor-gamma, sterol regulatory element binding protein-1c, fatty acid synthase, and fatty acid translocase, which are genes involved in lipid metabolism, were significantly downregulated in the ME group compared to the HF group. In contrast, the mRNA level of hepatic carnitine palmitoyl transferase-1 (involved in fatty acid oxidation) was upregulated by ME supplementation. Furthermore, administration of ME significantly downregulated the mRNA levels of inflammatory mediators such as hepatic tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), monocyte chemoattractant protein-1, and inducible nitric oxide synthase. The serum levels of TNF-α, IL-6, and nitric oxide were also significantly reduced in ME group compared to the HF group. Expression of hepatic miR-221 and miR-222, which increase in the inflammatory state of the liver, were also significantly inhibited in the ME group compared to the HF group. Conclusion: These results indicate that ME has the potential to improve hepatic lipid accumulation in high-fat diet-fed rats via modulation of inflammatory mediators and hepatic miR-221/222 expressions.