• Title/Summary/Keyword: Activity assay

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Rice Bran Phytic Acid Induced Apoptosis Through Regulation of Bcl-2/Bax and p53 Genes in HepG2 Human Hepatocellular Carcinoma Cells

  • Al-Fatlawi, Atheer Abbas;Al-Fatlawi, Anees Abbas;Irshad, Md.;Zafaryab, Md.;Alam Rizvi, M. Moshahid;Ahmad, Ayaz
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.8
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    • pp.3731-3736
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    • 2014
  • Phytic acid (PA) has been reported to have positive nutritional benefits and prevent cancer formation. This study investigated the anticancer activity of rice bran PA against hepatocellular carcinoma (HepG2) cells. Cytotoxicty of PA (0.5 to 4mM) was examined by MTT and LDH assays after 24 and 48h treatment. Apoptotic activity was evaluated by expression analysis of apoptosis-regulatory genes [i.e. p53, Bcl-2, Bax, Caspase-3 and -9] by reverse transcriptase-PCR and DNA fragmentation assay. The results showed antioxidant activity of PA in Fe3+ reducing power assay ($p{\leq}0.03$). PA inhibited the growth of HepG2 cells in a concentration dependent manner ($p{\leq}0.04$). After 48h treatment, cell viability was recorded 84.7, 74.4, 65.6, 49.6, 36.0 and 23.8% in MTT assay and 92.6, 77.0%, 66.8%, 51.2, 40.3 and 32.3% in LDH assay at concentrations of 1, 1.5, 2.0, 2.5, 3.0, and 3.5mM, respectively. Hence, treatment of PA for 24h, recorded viability of cells 93.5, 88.6, 55.5, 34.6 and 24.4% in MTT assay and 94.2, 86.1%, 59.7%, 42.3 and 31.6%, in LDH assay at concentrations of 1, 2.2, 3.0, 3.6 and 4.0mM, respectively. PA treated HepG2 cells showed up-regulation of p53, Bax, Caspase-3 and -9, and down-regulation of Bcl-2 gene ($p{\leq}0.01$). At the $IC_{50}$ (2.49mM) of PA, the p53, Bax, Caspase-3 and-9 genes were up-regulated by 6.03, 7.37, 19.7 and 14.5 fold respectively. Also, the fragmented genomic DNA in PA treated cells provided evidence of apoptosis. Our study confirmed the biological activity of PA and demonstrated growth inhibition and induction of apoptosis in HepG2 cells with modulation of the expression of apoptosis-regulatory genes.

Schisandra Chinensis Inhibits Oxidative DNA Damage and Lipid Peroxidation Via Antioxidant Activity

  • Jeong, Jin-Boo;Jeong, Hyung-Jin
    • Korean Journal of Plant Resources
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    • v.22 no.3
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    • pp.195-202
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    • 2009
  • Schisandra chinensis have been traditionally used in Asia for the treatment of dyspnea, cough, mouth dryness, spontaneous diaphoresis, nocturnal diaphoresis, nocturnal emission, dysentery, insomnia and amnesia. The purpose of this study is to evaluate the protective effects of Schisandra chinensis on oxidative DNA damage and lipid peroxidation induced by ROS in non cellular and cellular system. DPPH radical, hydroxyl radical and hydrogen peroxide scavenging assay were used to measure the antioxidant activities. Phi X-174RF I plasmid DNA cleavage assay and intracellular DNA migration assay were used to evaluate the protective effect on oxidative DNA damage. MTT assay and lipid peroxidation assay were used for evaluating the protective effect on oxidative cell damage. It was found to scavenge DPPH radical, hydrogen peroxide and hydroxyl radical and it inhibited oxidative DNA damage, lipid peroxidation and cell death induced by hydroxyl radical. These data indicate that Schisandra chinensis possesses a spectrum of antioxidant and DNA-protective properties

Application of cabbage Peroxidase for Glucose Assay (양배추 Peroxidase의 포도당 분석에의 이용)

  • Park, In-Shik;Kho, Sun-Ok;Nam, in
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.3
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    • pp.224-228
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    • 1990
  • Cabbage contained high peroxidase activity among tested plant sources. The cabbage peroxi-dase can replace horseradish peroxidase to assay glucose with glucose oxidase. The amount of glucose can be determined quantitatively by glucose oxidase-cabbage peroxidase. The opti-mum pH and temperature for enzymatic glucose determination by glucose oxidase-cabbage peroxidase were 6.0 and 35-45$^{\circ}C$ respectively. The glucose assay was inhibited by addition of various metal salts such as mercuric chloride lead acetate silver nitrate ammonium molyb-date sodium tunstate and cupric sulfate. The relationship between absorbance and amount of glucose was linear up to 8.33 mM glucose in the assay mixture under the assay conditions.

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Desmutagenicity of Enzymatically Browned Substances Obtained from the Reaction of Prunus salicina (Red) Enzyme and Polyphenols (재래종 적색자두(Prunus salicina) 효소갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi;Hong, Eun-Hee;Omura, Hirohisa
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.212-219
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    • 1987
  • The rec-assay on Bacillus subtilis strains H17$({Rec}^+)$ and M45$({Rec}^-)$, the Ames test with modification of preincubation on Salmonella typhimurium TA98 and TA100 and DNA-breaking test on double strand calfthymus DNA were carried out using enzymatically browned substances obtained from the reaction of Prunus salicina (Red) enzyme and polyphenols. The spore rec-assay of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone. 3,4-dihydrohyoluene and chlorogenic acid showed non-mutagenic activity The spore rec-assay showed a little influence of ${Zn}^{2+}$ and ${Ni}^{2+}$ on the action of four kinds of enzymatic browning reaction products. The enzymatic browning reaction products of polyphenols did not show DNAbreaking activity. ${Cu}^{2+}$ of various metal ions influenced on DNA-breaking of enzymatic browning reaction products of pyrogallol. However, enzymatic browning reaction products of chlorogenic acid inhibited on DNA-breaking activity. Four kinds of enzymatic browning reaction products showed non-mutagenic activity on Salmonella typhimurium TA98 and TA100 with S-9 mix. In the mutagenicity on Salmonella typhimurium TA98 and TA100 with S-9 mix in the presence of benzo$({\alpha})$pyrene which is the carcinogenic substances, four kinds of enzymatic browning reaction products showed desmutagenic activity.

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Efficacy of Safflower on the Acne Skin and Its Application for Facial Cleansing Biomedical Material (홍화잇꽃의 여드름피부 개선효과 및 세안용 생약식물소재 응용)

  • Park, Young-Ho;Lee, Chang-Seop
    • Journal of the Korean Chemical Society
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    • v.55 no.3
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    • pp.400-404
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    • 2011
  • Safflower is called as the 'beneficial flower' because 'it helps human health', and it was introduced as red flower in Tonguibogam due to the red color of floral leaf. From old times, it has been used for the material of cloth and rouge. Recently, polyphenol compound, the main ingredient of safflower, known as anti-aging and anti-oxidizing material in the healthy food industry becomes the emerging hot topic. This study aims to confirm by DDT (Disk Diffusion Test) assay, MTT assay, and NF-${\kappa}$B Luciferase activity inhibition assay in vitro that polyphenol compound, which is the main ingredient of safflower, has the anti-microbial efficacy to inhibit the growth of acne germs that make troubles for the teenagers or middle aged. Also it aims to evaluate its clinical efficacy on the acne skin, utilizing the facial cleansing cosmetic form of soap sample. This study can contribute to take a major step forward to the development of cosmetic soap for acne in the cosmeceutical industry.

The Effects of 5 kinds of Injinsaryung-San fractions on Cell Viability, Cell Cycle Progression and Fas-mediated Apoptosis of HepG2 Cells (인진사령산 분획물이 간세포활성, 세포주기 및 Fas-Mediated Apoptosis에 미치는 영향)

  • 고흥;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.21 no.3
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    • pp.174-185
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    • 2000
  • Objectives : This study was carried out to evaluate the effects of five fractions on cell viability, cell cycle progression and apoptosis. Methods : This study employed MTT assay, Cell cycle analysis, Cpp32 protease assay, DNA fragmentation assay and Quantitative RT-PCR analysis. Results : In MTT assay, the butanol fraction of Injinsaryung-San has showed magnificent viability, while the $H_2O$ fraction and ethylacetate fraction also showed higher viability than the control group. The $H_2O$ fraction of Injinsaryung-San has showed magnificent viability, and butanol fraction and ethylacetate fraction of Injinsaryung-San with etoposide have also showed higher viability than the only etoposide group. Cell cycle analysis showed that each fraction of Injinsaryung-San had no significant effect on the cell cycle. DNA fragmentation assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction carried inhibitory effects on apoptosis induction. Cpp32 protease activity assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction decreased Cpp32 protease activity, with the butanol fraction displaying greater effects. Quantitative RT-PCR showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction suppressed Fas and Bax genes, the butanol fraction increased BcI-2 gene, however no effect on Cpp32. Conclusions : The data shows that the butanol fraction of Injinsaryung-San increases the hepatocyte viability and has the heptocelluar protective effect by the suppression of apoptosis through gene regulation.

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The root extract of Paeonia lactiflora Pall inhibits the oxidative damage via its anti-oxidant activity

  • Yun, Ji Young;Jeong, Jin Boo;Eo, Hyun Ji;Kwon, Kun Woo;Hong, Se Chul;Jeong, Hyung Jin;Koo, Jin Suk
    • The Korea Journal of Herbology
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    • v.27 no.6
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    • pp.7-13
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    • 2012
  • Objectives : Reactive oxygen species (ROS) have been associated with pathogenic processes including carcinogenesis through direct effect on DNA directly and by acting as a tumor promoter. Therefore, it has been regarded that ROS may be a major target for cancer prevention. The root of Paeonia lactiflora pall (PL), a traditional Chinese herb, has been a component of effective prescriptions for treatment of liver disease. Also, there are some reports about the antioxidant activities of the extracts from PL. However, little has been known about the effects of PL against oxidative damage. This work aimed to elucidate the anti-oxidant effects of Paeonia lactiflora pall (PL) in the non-cellular system and cellular system. Methods : Antioxidant activities of PL were evaluated by hydroxyl radical scavenging assay and $Fe^{2+}$ chelating assay. Anti-oxidative effect of PL was evaluated by ${\varphi}X$-174 RF I plasmid DNA cleavage assay in non-cellular system. In addition, DNA migration assay, expression level of phospho-H2AX, MTT assay and lipid peroxidation assay were performed for evaluate the anti-oxidative effect of PL in cellular system. Results : PL had a dose-dependent hydroxyl radical scavenging and $Fe^{2+}$ chelating capacity. In addition, PL inhibited oxidative DNA and cell damage induced by hydroxyl radical in non-cellular system and cellular system. Conclusion : Taken together, P. lactiflora pall may be possible for the application to a potential drug for treating the oxidative diseases such as cancer.

Free Radical Scavenging Activity and Protective Ability of Methanolic Extract from Duchesnea indica Against Protein Oxidation and DNA Damage

  • Hu, Weicheng;Shen, Wei;Wang, Myeong-Hyeon
    • Preventive Nutrition and Food Science
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    • v.14 no.4
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    • pp.277-282
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    • 2009
  • The antioxidant potency of methanolic extract of Duchesnea indica (MDI; Indian strawberry) was investigated by employing various established in vitro systems, such as total phenolic content, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, reducing power assay, metal chelating assay, superoxide radical scavenging activity and protective ability of DNA damage and protein oxidation. MDI inhibited metal chelating by 75.57% at 2 mg/mL, scavenged 50% DPPH free radical at 29.13 ${\mu}$g/mL, and eliminated approximately 46.21% superoxide radical at the concentration of 1 mg/mL. In addition, MDI showed strong ability on reducing power, DNA damage protection and protein oxidation protection. Overall, results suggested that MDI might be beneficial as a potent antioxidant and effectively employed as an ingredient in food applications.

Synthesis of 2-Thio-4-aminopyrimidine Derivatives as Anti-cancer Agent

  • Lee, Sang-Hyo;Lee, Jin-Ho
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.105-112
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    • 2009
  • The screening of the anti-cancer activity of the chemical library provided 2-thio-4-aminopyrimidine as the initial hit. The confirmation of structure and biological effect of hit was performed by synthesis and biological evaluation. The optimization of hit was performed by derivatization of substituents while keeping the core structure. The evaluation of growth inhibitory activity was carried out using SRB assay against 6 human cancer cell lines and human fibroblast. The growth inhibitory activity of compounds showed substituent dependency and more than 5 compounds showed complete growth inhibition of cancer cell lines at 10 ${\mu}M$ concentration. Chemical library screening followed by synthetic modification provided possibility that 2-thio-4-aminopyrimidine can be used as a new scaffold for the development of anti-cancer agent.

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Glycosides from the Flower of Clerodendrum trichotomum (누리장나무 꽃의 배당체 성분)

  • Lee, Jong-Wook;Bae, Jong Jin;Kwak, Jong Hwan
    • Korean Journal of Pharmacognosy
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    • v.47 no.4
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    • pp.301-306
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    • 2016
  • Four phenylpropanoid glycosides and a monoterpene glycoside were isolated from the flower of Clerodendrum trichotomum. Structures of the isolated compounds were identified as acteoside (1), martynoside (2), leucosceptoside A (3), isoacteoside (4) and neohancoside A (5) by spectroscopic analysis. Compounds 1-4 were isolated from the flower of C. trichotomum for the first time. Compound 5 was first obtained from genus Clerodendrum as well as family Verbenaceae. The antioxidant activity of compounds 1-5 were evaluated by the DPPH free radical scavenging assay. Compounds 1-4 exhibited strong antioxidant activity.