• Title/Summary/Keyword: cell fractions

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Anti-inflammatory Effect of Ethanol Extract from Eupatorium japonicum (등골나물 추출물의 항염증 효과)

  • Lee, Han-Na;Lim, Do-Young;Lim, Soon-Sung;Kim, Jong-Dai;Yoon, Jung-Han
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.65-71
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    • 2011
  • Eupatorium japonicum belongs to a family of Asteraceae plants and flowers of E. japonicum have been consumed as a tea. In this study, we investigated whether E. japonicum extract inhibits lipopolysaccharide (LPS)-induced inflammatory responses in Raw264.7 macrophages. The cells were treated with various concentrations (0, 1, 2.5, 5, or 10 mg/L) of 70% ethanol extract from E. japonicum flowers (EJE) in Raw264.7 cells. LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production were inhibited by EJE up to 67% and 49% of these productions, respectively without any reduction of viable cell numbers. EJE reduced LPS-induced expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 proteins and their corresponding mRNA levels. Additionally, EJE decreased the levels of interleukin (IL)-6, IL-1${\beta}$, and tumor necrosis factor (TNF)-${\alpha}$ mRNA. EJE was further fractionated with water, butanol, ethylacetate (EA), hexane, or methylene chloride (MC). Among the resulting five fractions, EA and MC, respectively from EJE significantly inhibited LPS-induced NO production (each inhibition rate was 85.3% of 10 mg/L EA fraction and 97.2% of 10 mg/L MC fraction) without significant cytotoxicity in Raw264.7 cells. These results indicate that EJE exhibits powerful effects of anti-inflammation and can be developed as a potential anti-inflammatory agent.

Fenofibrate decreases radiation sensitivity via peroxisome proliferator-activated receptor ${\alpha}$-mediated superoxide dismutase induction in HeLa cells

  • Liu, Xianguang;Jang, Seong-Soon;An, Zhengzhe;Song, Hye-Jin;Kim, Won-Dong;Yu, Jae-Ran;Park, Woo-Yoon
    • Radiation Oncology Journal
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    • v.30 no.2
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    • pp.88-95
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    • 2012
  • Purpose: The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) ${\alpha}$ and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofibrate (FF). Materials and Methods: Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. Results: In HeLa cells total SOD activity was increased with increasing FF doses up to 30 ${\mu}M$. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, $PPAR{\alpha}$ and $PPAR{\gamma}$ were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and $PPAR{\alpha}$ were not increased with FF. However, the mRNA of $PPAR{\gamma}$ was increased with FF. Conclusion: FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with $PPAR{\alpha}$.

Antitumor Activity of Methylene Chloride Fraction from Angelica Keiskei Through Induction of Apoptosis in Human Prostate Carcinoma DU145 Cells (전립선 암세포주 DU145의 세포고사 유도를 통한 신선초 (Angelica keiskei) 메틸렌 클로라이드 추출물의 항암효과)

  • Kang, Yoon-Mook;Kim, Sung-Moo;Kim, Hyun-Jung;Park, Kyung-Ran;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok;Ahn, Kwang-Seok
    • Journal of Korean Traditional Oncology
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    • v.15 no.1
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    • pp.19-27
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    • 2010
  • The roots and leaves of Angelica keiskei (AK) have been used for the treatment of various diseases including coronary heartdisease, hypertension, and cancer in the Korean folk medicine. However, the mechanism by which methylenechloride fraction (MF) from AK exerts anti-tumorigenic activity in human prostate cancer cells has not been fully understood. In the present study, we report the MF exerted the highest cytotoxicity against prostate cancer DU145 cells compared with other fractions. Especially, MF caused the accumulation of sub-G1 DNA contents of cell cycle and increased annexin V-positive apoptotic bodies and DNA fragmentation. MF down-regulated several proliferative (Cyclin D1) and anti-apoptotic (Bcl-xl, Bcl-2, IAP-1/2, and survivin)gene products in these cells. Hence, MF induced apoptosis through the caspase-3 activation in DU145 cells. We further confirmed that caspase-3 plays an importance role in MF-induced apoptosis in DU145 cells by using caspase-3 inhibitor. Additionally, we observed that MF potentiated Dox-induced apoptosis in DU145 cells. Taken together, our data demonstrate the evidence that MF induces apoptosis depend on caspase-3 activation of and overcomes resistance to chemotherapy in human prostate cancer cells.

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Insulin Sensitizing and Insulin-like Effects of Water Extracts from Kalopanax pictus NAKAI in 3T3-L1 Adipocyte (엄나무 추출물이 3T3-L1 지방세포에서 인슐린 민감성과 인슐린 유사성 작용에 미치는 영향)

  • Kim, Ho-Kyoung;Park, Sun-Min;Ko, Byoung-Seob
    • Applied Biological Chemistry
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    • v.45 no.1
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    • pp.42-46
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    • 2002
  • Effects of the water extract from Kalopanax pictus NAKAI on insulin-like action and glucose uptake in 3T3-L1 cells were investigated. The bark of K. pictus NAKAI was treated with hot water and the extract was freeze-dried. Total extract of K. pictus NAKAI was fractionated into 6 fractions with increasing gradients from 0 to 100% MeOH on Amberlite XDA-4. Treatment of 3T3-L1 fibroblasts with 1 ${\mu}g/ml$ and 10 ${\mu}g/ml$ of K. pictus NAKAI total extracts significantly increased the differentiation of the cells. When co-treated with inducers such a dexamethasone, 1-methyl-3-isobutylxanthine and insulin, the differentiation was increased at 1 ${\mu}g/ml$ of total extract, but not at 10 ${\mu}g/ml$. In 3T3-L1 adipocytes, glucose uptake was increased by 3.3 times with addition of 0.3 and 3 ${\mu}g/ml$ of Fr. 1 (0-10% MeOH) and Fr. 3 (30% MeOH) at 3 ng/ml insulin. In conclusion, K. pictus NAKAI contains such compounds that play a role of insulin-like action and insulin sensitizer.

Investigation on sample throughput of large scale splitter-less gravitational SPLITT fractionation (GSF) (Large scale Gravitational SPLITT Fractionation (GSF)에서의 시료 throughput에 관한 연구)

  • Choi, Hyo Jae;Kim, Woon Jung;Eum, Chul Hun;Lee, Seungho
    • Analytical Science and Technology
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    • v.26 no.1
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    • pp.34-41
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    • 2013
  • Split-flow thin cell (SPLITT) fractionation (SF) is a rapid separation technique capable of separating colloidal particles or macromolecules into two or more fractions. SF allows fractionations in a preparative scale as sample is fed continuously. Generally SF uses a thin ribbon-like channel equipped with two flow stream splitters at the inlet and outlet of the channel. Thus there exist two flow inlets and two flow outlets at the top and bottom of the inlet and outlet of the channel, respectively. There are two operating modes in SF, conventional mode and full-feed mode (FFD). Although the resolution in the FFD mode is lower than that in the conventional mode, FFD mode has some merits. The design of the channel and operation are simpler in the FFD mode, as it does not require the feeding of the solvent. Thus there is no flow stream splitter at the channel inlet, and only one pump is needed, unlike the conventional mode, where two pumps are required for the feedings of the sample and the solvent separately. Also the sample is not diluted in the FFD mode as there is no solvent feeding, which is important for fractionation samples with low colloidal concentrations such as environmental samples. For some of environmental samples, pre-concentration is often required. In this study, a new large-scale splitter-less FFD-SF channel was implemented, where there is no splitter at the outlet as well as at the inlet of the channel. It was possible to build the channel in a much larger dimension than conventional ones, allowing much higher sample throughput (TP). The new channel was tested and optimized with polyurethane (PU) latex beads, and then applied to large-scale separation of Polyacrylate (PA).

Effects of Gonadotropins, Prostaglandin $F_{2{\alpha}}$, and Ouabain on the $Ca^{++}-ATPase$ Activity in Luteal Membranes (Gonadotropins, Prostaglandin $F_{2{\alpha}}$ 및 Ouabain이 황체막의 $Ca^{++}-ATPase$ 활성도에 미치는 영향)

  • Koo, Bon-Sook;Kim, In-Kyo
    • The Korean Journal of Physiology
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    • v.21 no.1
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    • pp.47-58
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    • 1987
  • It has been reported that the luteal function may be regulated by the intracellular $Ca^{++}$ level which may be adjusted partially by the high affinity $Ca^{++}-ATPase$ in luteal cell membranes. Then, one may expect that luteotropic and/or luteolytic agents, such as gonadotropins, prostaglandin $F_{2{\alpha}}\;(PGF_{2{\alpha}})$ and ouabain, affect the intracellular $Ca^{++}$ level. In this present study, therefore, we examined the effects of luteinizing hormone (LH, or human chorionic gonadotropin, hCG), $PGF_{2{\alpha}}$ and ouabain on the kinetic properties of the high affinity $Ca^{++}-ATPase$ in light membrane, heavy membrane, and microsomal fractions from the highly luteinized ovary. LH (or hCG) increased the affinity and the Vmax for $Ca^{++}$ both in light membrane and heavy membrane. $PGF_{2{\alpha}}$ increased the Vmax in light membrane and decreased the Km in heavy membrane for $Ca^{++}$ at low concentration $(5\;{\mu}g/ml)$. At higher concentration, however, $PGF_{2{\alpha}}$ oppositly affected on kinetic properties, that shown at low concentration. Ouabain, a potent inhibitor of $Na^+-K^+-ATPase$, increased the Km at high concentration $(10^{-4}\;M)$, however, decreased the Vmax for $Ca^{++}$ in light membrane at low concentration $(10^{-6}\;M)$. Also, ouabain increased the Km for $Ca^{++}$ in heavy membrane without changes in the Vmax at both concentrations. It seems that LH and low dose of $PGF_{2{\alpha}}$ increase the intracellular $Ca^{++}$ level and cause in activation of $Ca^{++}-ATPase$, however, higher dose of $PGF_{2{\alpha}}$ and ouabain inhibit directly $Ca^{++}-ATPase$ activity and result in increase in intracellular $Ca^{++}$ level. According to the above results, we suggest that luteotropic and/or luteolytic agents regulate the luteal progesterone $(P_4)$ production through two different pathways; one is cyclic adenosine monophosphate (cAMP)-dependent and another is $Ca^{++}-dependent$. Intracellula. $Ca^{++}$ level regulated by the high affinity $Ca^{++}-ATPase$ may affect both pathways in a time-dependent fashion. LH (or hCG) acts on the luteal $P_4$ production via both pathways. The initial step is $Ca^{++}$ dependent, and the late step is cAMP dependent. $PGF_{2{\alpha}}$ and ouabain increase the intracellular $Ca^{++}$ concentration so that basal luteal $P_4$ production is increased and LH-stimulated $P_4$ production is inhibited by the inhibiting LH-dependent adenylate cyclase activity.

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Anticancer and Immunopotentiating Activities of Crude Polysaccharides from Pleurotus nebrodensis on Mouse Sarcoma 180

  • Cha, Youn Jeong;Alam, Nuhu;Lee, Jae Seong;Lee, Kyung Rim;Shim, Mi Ja;Lee, Min Woong;Kim, Hye Young;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Tae Soo
    • Mycobiology
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    • v.40 no.4
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    • pp.236-243
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    • 2012
  • Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. ${\beta}$-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of $10{\sim}2,000{\mu}g/mL$. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of $50{\mu}g/mL$, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at $200{\mu}g/mL$. Maximum production of nitric oxide ($14.3{\mu}M$) was recorded in the Fr. NaCl fraction at $200{\mu}g/mL$. Production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-${\alpha}$, IL-$1{\beta}$, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

Antimicrobial Activity of Extracts and Fractions of Ginkgo biloba Leaves, Seed and Outer Seedcoat (은행 잎, 종실 및 외종피 추출물의 항균활성)

  • Park, Saet-Byoul;Cho, Gyu-Seong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.7-13
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    • 2011
  • This study was conducted to investigate the antimicrobial activity of Ginkgo biloba L. leaves, seed and outer seedcoat against bacteria. Antimicrobial effects of Ginkgo biloba L. leaves (GBL), seed (GBS) and outer seedcoat (GBO) were examined by paper disc method and optical density method to determine minimum inhibition concentration (MIC), and observed by scanning electron microscope (SEM) to figure out the morphological change on the surface when Ginkgo biloba leaves extract was treated. The extracts of GBL, GBS and GBO were extracted by solvents such as methanol, ethanol, water. The methanol extract of GBL and GBO showed the highest antimicrobial activity against Bacillus cereus, Bacillus subtilis, Klebsiella pneumoniae, Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, Yersinia enterocolitica except Escherichia coli and thus was further fractionated. The MICs of the chloroform fraction of GBL methanol extract were $125{\mu}g$/mL against B. subtilis, and L. monocytogenes; GBO methanol extract were $62.5{\mu}g$/mL against B. cereus and $125{\mu}g$/mL against B. subtilis, and L. monocytogenes. The microorganisms were treated with chloroform extracts ($2000{\mu}g$/mL) of GBL and GBO methanol extracts. It was observed by scanning electron microscope (SEM). The cells were expanded and a part of cell wall was completely destructed by GBL and GBO. Thus Ginkgo biloba L. leaves and outer seedcoat could be further developed into a natural antimicrobial agent.

Excess Molar Enthalpies and Excess Molar Volumes for the Binary Mixtures {1,2-dichloropropane+2-(2-methoxyethoxy)ethanol, and +2-(2-ethoxyethoxy)ethanol} at 298.15 K (2성분계 {1,2-dichloropropane+2-(2-methoxyethoxy)ethanol 및 + 2-(2-ethoxyethoxy)ethanol}에 대한 298.15 K에서의 과잉몰엔탈피 및 과잉몰부피)

  • Kim, Jaewon;Kim, Moongab
    • Korean Chemical Engineering Research
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    • v.44 no.5
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    • pp.444-452
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    • 2006
  • This paper reports experimental excess molar volumes $V^E_m$ using a digital vibrating-tube densimeter and excess molar enthalpies $H^E_m$ by means of an isothermal microcalorimeter with a flow mixing cell for the binary mixtures{1,2-dichloropropane + 2-(2-methoxyethoxy)ethanol} and {1,2-dichloropropane + 2-(2-ethoxyethoxy)ethanol} at 298.15 K under atmospheric pressure. All the $V^E_m$ and $H^E_m$ of the two binary mixtures showed S-shaped forms, being negative for poor and positive for rich 1,2-dichloropropane mole fractions. These show that the excess properties were shown to be negative deviation from ideality due to the strong self-association effect among 2-(2-alkoxyethoxy)ethanol molecules at an early stage of mixing, a relatively high energy then is needed to break hydrogen bonds of 2-(2-alkoxyethoxy)ethanol with an increase ofhalogenated hydrocarbon molecular at high mole fraction of 1,2-dichloropropane. The values of excess molar properties($V^E_m$ and $H^E_m$) were fitted by the Redlich-Kister equation using Nelder-Mead's simplex pattern search method. The Wilson, NRTL, and UNIQUAC models were used to correlate the $H^E_m$ values.

Taxifolin Inhibited the Nitric Oxide Production and Expression of Pro-inflammatory Cytokine mRNA in Lipopolysaccharide-stimulated RAW264.7 Cells

  • Rhee, Man-Hee;Endale, Mehari;Kamruzzaman, SM;Lee, Whi-Min;Park, Hwa-Jin;Yoo, Myung-Jo;Cho, Jae-Youl
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.147-155
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    • 2008
  • In previous works, we found that solvent extract of Opuntia humifusa Raf., a member of the lactaceae family, displayed potent anti-oxidative and anti-inflammatory activities. Thus, all solvent fractions, except for the water layer, showed potent scavenging effects. According to activity-guided fractionation, one of active radical scavenging principles in the ethyl acetate fraction was found to be taxifolin. In this study, we investigated whether taxifolin showed anti-oxidative activity. In addition, taxifolin modulated nitric oxide (NO) release and the expression of pro-inflammatory cytokine mRNA such as interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-${\alpha}$. Taxifolin showed potent anti-oxidant activity with the $IC_{50}\;of\;8.5{\pm}1.4\;and\;9.3{\pm}1.0{\mu}M$ using xanthine/xanthine oxidase (XO) assay and 2,2-Diphenyl-lpicrylhydrazyl radical (DPPH) assay, respectively. We next determined the role of taxifolin on the immunomodulating activity using murine macrophage cell line RAW264.7 cells. Taxifolin dose-dependently inhibited NO production in lipopolysaccharide (LPS)-activated RAW264.7. It also significantly blocked the expression of inducible NO synthase (iNOS) mRNA in the LPS-stimulated RAW264.7 cells. In addition, taxifolin potently suppressed the expression of IL-$1{\beta}$, IL-6 and GM-CSF mRNA in LPS-activated RAW264.7 cells, but not that of TNF-${\alpha}$ Moreover, taxifolin significantly inhibited the transcriptional activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and activator protein -1 (AP-1). These results suggest that taxifolin may downregulate inflammatory iNOS, IL-$1{\beta}$, IL-6 and GM-CSF gene expressions through inhibition of NF-K and AP-1 activation in LPS-stimulated RAW264.7 cells.

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