• Title/Summary/Keyword: human activity

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Mitochondria-Targeted Apoptosis in Human Cytomegalovirus-Infected Cells

  • Lee, Gyu-Cheol;Lee, Jae Ho;Kim, Bo Yeon;Lee, Chan Hee
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1627-1635
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    • 2013
  • Mitochondria often play central roles in apoptotic pathways, and disruption of the mitochondrial transmembrane potential (${\Delta}{\psi}m$) has been observed in various cells undergoing apoptosis. Human cytomegalovirus (HCMV) infection induces apoptosis in permissive cells; however, investigations of mitochondria-targeted apoptosis in HCMV-infected human foreskin fibroblast (HFF) cells have been limited. Here, we investigated the mitochondrial apoptosis pathway in HCMV-infected HFF cells. Flow cytometry analysis using JC-1 revealed that HCMV infection induces disruption of ${\Delta}{\psi}m$ in HFF cells when administered 24 h post-infection (hpi), and this disruption was maximized at 48 hpi. Moreover, cytochrome c, normally a mitochondrial inner membrane protein, was detected in cytoplasmic extracts of HCMV-infected cells, but not mock-infected cells, by western blot analysis at 24 hpi. A caspase activity assay based on fluorescence spectrophotometry using a fluorogenic substrate revealed an increase in caspase-3 activity at 48 hpi in HCMV-infected cells. Caspase-8 activity was increased at 72 hpi in HCMV-infected cells. These results imply that HCMV infection induces mitochondria-mediated apoptosis in HFF cells.

Cytochrome P-450 3A4 proximal promoter activity by histone deacetylase inhibitor in HepG2 cell.

  • Kim, Ja-Young;Ahn, Mee-Ryung;Sheen, Yhun-Yhong
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.88-88
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    • 2003
  • Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

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Quality Characteristics and Antioxidant Activities of Cookies added with Purple Sweet Potato Powder (자색고구마 분말 첨가량을 달리한 쿠키의 항산화 활성 및 품질 특성)

  • Liu, Ya-Nan;Jeong, Da-Hye;Jung, Ji-Hye;Kim, Hyun-Sook
    • Korean journal of food and cookery science
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    • v.29 no.3
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    • pp.275-281
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    • 2013
  • This study was performed to evaluate the quality characteristics and antioxidative activities of cookies prepared with different amounts of purple sweet potato powder. Moisture and crude ash contents were lowest in the control group followed in order by the 10%, 20%, and 30% purple sweet potato powder addition groups. The lightness and yellowness values measured by Hunter color system decreased based on the amount of purple sweet potato powder concentration added to cookies. The total phenolic compound and anthocyanin contents were the highest in 30% purple sweet potato powder added cookies. The DPPH radical scavenging activity of 30% purple sweet potato powder added cookies showed the highest values among groups. In the sensory evaluation, the scores for taste, texture and overall preference were significantly highest in cookies with 10% and 20% purple sweet potato powder. Considering the scores of antioxidant activity and sensory evaluation, the concentration of between 10% and 20% addition of purple sweet potato powder would be ideal for development of functional cookies.

Ethyl Acetate Extract from Tissue-Cultured Mountain Ginseng Adventitious Roots Inhibits In Vitro Platelet Aggregation in Whole Human Blood and Augments Peripheral Blood Flow in Mice

  • Lee, In-Sun;Kim, Seul-Ki;Jeon, Min-Hwa;Jeon, Won-Kyung
    • Journal of Ginseng Research
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    • v.35 no.4
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    • pp.442-448
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    • 2011
  • We previously reported that in vitro anti-platelet activity of tissue-cultured mountain ginseng (TCMG) ethanol extracts show improved efficacy when compared with commercial ginseng products such as Korean red ginseng and Panax ginseng. However, information on the anti-platelet activity of the ethyl acetate fraction from TCMG adventitious roots is limited. Therefore, in this study, we further investigated the effects of an ethyl acetate extract of TCMG (EA-TCMG) adventitious roots on in vitro antiplatelet activity in whole human blood and its effect on peripheral blood flow in mice. We found that EA-TCMG inhibited platelet aggregation with $IC_{50}$ values of 271, 180, and 147 ${\mu}g$/mL induced by collagen, adenosine-5'-diphosphate, and arachidonic acid, respectively. Among the three agonists used, thromboxane $A_2$ formation induced by arachidonic acid was markedly suppressed. Furthermore, EA-TCMG improved the peripheral circulatory disturbance by improving vascular blood flow. In conclusion, these results suggest that ethyl acetate extracts from TCMG adventitious roots might inhibit vascular platelet aggregation and thrombus formation.

Involvement of PI3K and MMP1 in PDGF-induced Migration of Human Adipose-derived Stem Cells

  • Lim, Yoonhwa;Lee, Minji;Jeong, Hyeju;Kim, Haekwon
    • Development and Reproduction
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    • v.21 no.2
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    • pp.167-180
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    • 2017
  • Human adult stem cells have widely been examined for their clinical application including their wound healing effect in vivo. To function as therapeutic cells, however, cells must represent the ability of directed migration in response to signals. This study aimed to investigate the mechanism of platelet-derived growth factor (PDGF)-induced migration of the human abdominal adipose-derived stem cells (hADSCs) in vitro. A general matrix metalloproteinase (MMP) inhibitor or a MMP2 inhibitor significantly inhibited the PDGF-induced migration. PDGF treatment exhibited greater mRNA level and denser protein level of MMP1. The conditioned medium of PDGF-treated cells showed a caseinolytic activity of MMP1. Transfection of cells with siRNA against MMP1 significantly inhibited MMP1 expression, its caseinolytic activity, and cell migration following PDGF treatment. Phosphatidylinositol 3-kinase (PI3K) inhibitor reduced the migration by about 50% without affecting ERK and MLC proteins. Rho-associated protein kinase inhibitor mostly abolished the migration and MLC proteins. The results suggest that PDGF might signal hADSCs through PI3K, and MMP1 activity could play an important role in this PDGF-induced migration in vitro.

Cytotoxic Effects of Strawberry, Korean Raspberry, and Mulberry Extracts on Human Ovarian Cancer A2780 Cells

  • Lee, Dahae;Kang, Ki Sung;Lee, Sanghyun;Cho, Eun Ju;Kim, Hyun Young
    • Preventive Nutrition and Food Science
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    • v.21 no.4
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    • pp.384-388
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    • 2016
  • Reactive oxygen species are tumorigenic by their ability to increase cell proliferation, survival, and cellular migration. The purpose of the present study was to compare the antioxidant activity and cytotoxic effects of 3 berry extracts (strawberry, Korean raspberry, and mulberry) in A2780 human ovarian carcinoma cells. Except for raspberry, the ethyl acetate or methylene chloride fractions of berries containing phenolic compounds exerted dose dependent free radical scavenging activities. In the raspberry fractions, the hexane fraction also exhibited potent antioxidant activity. The cytotoxic effects of berries extracts in A2780 human ovarian carcinoma cells were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Surprisingly, co-treatment with n-butanol (BuOH) fractions of berries showed stronger cytotoxic effects compared to the other fractions. These findings suggest that potent anticancer molecules are found in the BuOH fractions of berries that have stronger cytotoxic activity than antioxidants.

Cytochrome P-450 3A4 proximal promoter activity by histone deacetylase inhibitor in HepG2 cell.

  • Kim, Ja-Young;Ahn, Mee-Ryung;Sheen, Yhun-Yhong
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.178-178
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    • 2003
  • Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this Is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

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Inhibitory Activity of Edible Plant Extracts on Proliferation of Human Umbilical Vein Endothelial Cells (HUVECs)

  • Song, Myoung-Chong;Kim, Sung-Hoon;Kwak, Ho-Young;Yang, Hye-Joung;Bang, Myun-Ho;Chung, In-Sik;Lee, Youn-Hyung;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.249-253
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    • 2007
  • Thirteen edible plants previously reported to show inhibitory activities on farnesyl protein transferase (FPTase) and phosphatase of the regenerating liver-3 (PRL-3) were evaluated for inhibitory activity on the proliferation of human umbilical vein endothelial cells (HUVECs). Four plant extracts, Oenothera erythrosepala, Perilla frutescens, Panicum miliaceum, and Quercus acutissima, significantly inhibited the proliferation of HUVECs induced by the basic fibroblast growth factor (bFGF) without cytotoxicity at 100 ${\mu}g/mL$. Myristica fragrans, Rosmarinus officinalis, and Syringa patula also showed inhibitory activity on the proliferation with only mild cytotoxicity.

Effects of a xenographic bovine bone on the bone mineralization in human fetal osteoblasts (우골 유도 합성골이 사람 태아 골모세포의 골 광물화 과정에 미치는 영향)

  • Sun, Ki-Jong;Hyun, Ha-Na;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.32 no.4
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    • pp.801-809
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    • 2002
  • The ultimate goal of periodontal therapy is to promote the regeneration of lost periodontal tissue, there have been many attempts to develop a method to achieve this goal, hut none of them was completely successful. The purpose of this study is to evaluate the effects of Bio-Oss(R) on alkaline Phosphatase (ALP) activity in human fetal osteoblasts (hFOB1). The results of this study were as follows, in ALP Activity, 100 ${\mu}g/ml$ Bio-Oss(R) treated group showed significantly increased value than negative control group, but positive group($10^{-7}$ M dexamethasone treated group) showed the highest ALP activity at 3 day. In mineralization assay, numerous mineralized nodules were identified as darkly stained spots in 100${\mu}g/ml$ Bio-Oss(R) treated group than two control groups, whereas a small number of mineralized nodules were showed in the positive control. ALP may relate to the initial phase of bone nodule formation. On the basis of these results, this study showed Bio-Oss(R) is capable of accelerating new bone formation through hFOBl differentiation in vitro.

Metabolic Activity of Desalted Ground Seawater of Jeju in Rat Muscle and Human Liver Cells

  • Kim, Bo-Youn;Lee, Young-Ki;Park, Deok-Bae
    • Fisheries and Aquatic Sciences
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    • v.15 no.1
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    • pp.21-27
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    • 2012
  • Ground seawater in the east area of the volcanic Jeju Island contains abundant minerals. We investigated the metabolic activity of electrodialyzed, desalted ground seawater (EDSW) from Jeju in both cultured cells and animals. The addition of EDSW to the culture medium (up to 20%, v/v) reduced the leakage of lactate dehydrogenase and increased MTT activity in CHO-IR cells. EDSW (10%) promoted insulin-induced glucose consumption in L6 muscle cells as well as the activities of the liver ethanol-metabolizing enzymes, alcohol dehydrogenase and aldehyde dehydrogenase. Moreover, EDSW suppressed palmitate-induced intracellular fat accumulation in human hepatoma $HepG_2$ cells. Activities of AMP-stimulated protein kinase and acetyl CoA carboxylase, enzymes that modulate fat metabolism, were altered by EDSW in $HepG_2$ cells toward the suppression of intracellular lipid accumulation. EDSW also suppressed hepatic fat accumulation induced by a high-fat diet in mice. Taken together, EDSW showed beneficial metabolic effects, including the enhancement of ethanol metabolism and insulin-induced glucose consumption, and the suppression of intrahepatic fat accumulation.