• Title/Summary/Keyword: HEP method

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Effects of Hesperidine, Naringin and Their Aglycones on the In Vitro Activity of Phosphatidate Phosphohydrolase, and on the Proliferation and Growth in Cultured Human Hepatocytes HepG2 Cells (In Vitro 에서 Phosphatidate Phosphohydrolase 활성과 HepG2 세포증식에 미치는 Hesperidine, Naringin 및 그 Aglycone Flavonoid의 영향)

  • Cha, Jae-Young;Cho, Young-Su
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.577-582
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    • 1997
  • Effects of four citrus flavonoids, hesperidin, naringin and their aglycones on phosphatidate phosphohydrolase(PAP, EC 3.1.3.3) activity were examined using isolated rat microsomes as an enzyme source. In addition, these flavonoids were tested to see whether they exert any influence on the proliferation and growth in cultured human hepatocytes HepG2 cells. Flavonoids at concentration up to $10{-4}M$ had no significant effect on the number of cells and cell proliferation by MTT cell growth assay method, whereas aglycone flavonoids, hesperetin and narigenin, at concentration of $10{-3}M$ significantly inhibited cell proliferation. Hesperetin inhibited PAP activity in a dose-dependent manner starting at concentration of $10{-5}M$. Narigenin at concentration of $10{-2}M$ inhibited PAP activity markedly, while the other flavonoids did not show any significant effect. The present study, therefore, demonstrated that aglycone flavonoids exerted portent effects on PAP activity and on cell proliferation.

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Inhibitory Effects of Naringenin, Kaempherol, and Apigenin on Cholesterol Biosynthesis in HepG2 and MCF-7 Cells

  • Kim, Kee-Tae;Yeo, Eun-Ju;Moon, Sun-Hee;Cho, Ssang-Goo;Han, Ye-Sun;Nah, Seung-Yeol;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1361-1364
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    • 2008
  • The inhibitory effects of naringenin, kaempherol, and apigenin on the production of cholesterol in HepG2 KCLB 88065 and MCF-7 KCLB 30022 cells were evaluated. In this study, quercetin was used as a reference reagent. After incubation for 3 days, fat-soluble contents of both cell types were extracted by using the Folch method and the cholesterol contents in both cultured cells were determined by high performance liquid chromatography. The concentration of cholesterol in untreated each tissue cells was $12.2{\pm}0.11$ and $8.83{\pm}0.12\;mg/g$ of lipid, respectively. The total concentration of each flavonoid was adjusted to 0, 35, or $350{\mu}M$ in the culture broth. As the results, the addition of 2% methanol and dimethyl sulfoxide (DMSO) to the media (control for flavonoid solvents) did not significantly affect cell growth; however, DMSO caused an increase in the production of cholesterol. Each flavonoid inhibited the production of cholesterol in both HepG2 and MCF-7 cells at the concentration of $35{\mu}M$ above. In addition, the inhibitory effect of kaempherol on the production of cholesterol in these cells was greater than the other flavonoids tested and HepG2 cells are more sensitive to flavonoids than MCF-7. From the results, the inhibitory effects of flavonoids on cholesterol production are different depending on the cell type.

Studies on the Pathogenic Test of Yersinia enterocolitica (Yersinia enterocolitica 의 병원성 검정에 관한 연구)

  • Lim, Soon-Young;Yoon, Suk-Kwon
    • Korean Journal of Food Science and Technology
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    • v.33 no.4
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    • pp.486-491
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    • 2001
  • The pathogenicity for one hundred strains of domestic and foreign Y. enterocolitica was tested with HEp-2 cell invasion method as a reference. The serotyping, biotyping, PCR and esculin hydrolyis, salicin fermentation, pyrazinamidase activity, indole production, xylose fermentation, CRMOX and autoagglutination were compared to determine the possibility of pathogenic detection method. According to the test results, serotyping was limited to verify pathogenicity, however, biotyping was quite related to pathogenicity up to 99%. The biotype 1A strains were non-pathogenic, while all strains of biotype $1B{\sim}4$ showed pathogenicity with the exception of one strain belonged to type 1B. The esculin and salicin test results were completely close and correlated to pathogenicity up to 99%. The HEp-2 cell invasion and pyrazinamidase test were related to pathogenicity by 95%. Biochemical tests such as D-xylose fermentation, CRMOX agar test and autoagglutination in broth were effective as a support test. It is strongly recommended that sequencial esculin test and PCR test could be done to verify pathogenicity of Y. enterocolitica as the easiest and accurate procedure.

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Killing Effects of Different Physical Factors on Extracorporeal HepG2 Human Hepatoma Cells

  • Zhang, Kun-Song;Zhou, Qi;Wang, Ya-Feng;Liang, Li-Jian
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.1025-1029
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    • 2012
  • Objective: To determine the killing effects on extracorporeal HepG2 cells under different temperatures, pressures of permeability and lengths of treatment time. Method: According to different temperatures, pressures of permeability and lengths of treating time, extracorporeal HepG2 cells of human hepatoma cell-line were grouped to 80 groups. Cell index (CI) as the measurement of killing effect were calculated by monotetrazolium (MTT) methods, i.e., CI =1- (the OD value in treated group - the OD value in blank control group) / (mean of untreated control group - mean of blank control group). According to the factorial design, data were fed into SPSS 10.0 and analyzed by three-way ANOVA (analysis of variance). Result: Temperature, pressure of permeability and length of treating time all had effects on the CI (cell index) level. Length of treating time was the most influential factor of the three. Additionally, any two of them all had statistically significant interactive effects on the CI level. When treated for 5-30 min, destilled water at $46^{\circ}C$ stably generated the highest CI. Conclusion: The "$46^{\circ}C$-destilled water-60 min" was considered as the optimal combination of conditions which lead to highest CI. We suggest exerting celiac lavage for 15 min with stilled water at $40^{\circ}C-43^{\circ}C$ in surgical practice as a hyperthermia treatment to achieve ideal killing effects on free cancer cells, which is feasible, practical, and clinically effective.

Purification and Identification of Cytotoxic Compounds from the Root of Rumex crispus L. (소리쟁이 뿌리로부터 세포독성물질 분리 및 정제)

  • Cho, Yong Beom;Kim, Jae Yeon;Kwon, Nam Woo;Hwang, Bang Yeon;Kim, Jun Gu;Woo, Sun Hee;Lee, Moon Soon
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.3
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    • pp.208-217
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    • 2019
  • Background: In the present study, we identified two cytotoxic compounds from the root of Rumex crispus L. using a bioassay-based method. Methods and Results: Compared with the other fractions, the diethyl ether ($Et_2O$) fraction of R. crispus root extract exhibited the strongest of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect [scavenging concentration 50% $(SC_{50})=63.8{\pm}1.47{\mu}g/m{\ell}$], nitric oxide (NO) production inhibitory effect on the mouse macrophage cell line RAW264.7 [inhibitory concentration 50% $(IC_{50})=60.9{\pm}7.52{\mu}g/m{\ell}$] and cytotoxicity effect on the human hepatoma cell line, HepG2 [lethal concentration 50% $(LC_{50})=115.4{\pm}1.86{\mu}g/m{\ell}$]. According to the bioassay-based method, two cytotoxic compounds were purified from the $Et_2O$ fraction by using column chromatography and preparative high performance liquid chromatography (prep-HPLC). These two compounds were identified as parietin and chrysophanol by using nuclear magnetic resonance (NMR) and liquid chromatography quadruple time of flight mass spectrometry (LC-QTOF-MS). In addition, both parietin and chrysophanol exhibited a cytotoxicity effect on HepG2 cells, their $LC_{50}$ values were $169.1{\pm}17.67{\mu}M$ and $111.5{\pm}6.62{\mu}M$, respectively. Conclusions: Parietin and chrysophanol isolated from the $Et_2O$ fraction of the R. crispus root extract showed cytotoxicity in HepG2 cell.

A study on the forecasting of container cargo volumes in northeast ports by development of competitive model (컨테이너 항만간의 경쟁 상황을 고려한 물동량예측에 관한 연구)

  • K.T.Yeo;Lee, C.Y.
    • Proceedings of the Korean Institute of Navigation and Port Research Conference
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    • 1998.10a
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    • pp.263-269
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    • 1998
  • The forecasting of container cargo volumes should be estimated correctly because it has a key roles on the establishment of port development planning, and the decision of port operating system. Container cargo volumes have a dynamic characteristics which was changed by effect of competitive ports. Accordingly forecasting was needed overall approach about competitive port's development, alternation and information. But, until now, traffic forecasting was not executed according to competitive situation, and that was accomplished at the point of unit port. Generally, considering the competition situation, simulation method was desirable at forecasting because system's scale was increased, and the influence power was intensified. In this paper, considering this situation, the objectives can be outlined as follows. 1) Structural model constructs by System dynamics method. 2) Structural simulation model develops according to modelling of competitive situation by expended SD method which included HEP(Hierarchical Fuzzy Process) And actually, effectiveness was verified according to proposed model to major port in northeast asia.

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Identification of Differentially Expressed Genes (DEGs) by Malachite Green in HepG2 Cells

  • Kim, Youn-Jung;Song, Mee;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.1
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    • pp.22-30
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    • 2008
  • Malachite Green (MG), a toxic chemical used as a dye, topical antiseptic and antifungal agent for fish, is highly soluble in water, cytotoxic to various mammalian cells and also acts as a liver tumor promoter. In view of its industrial importance and possible exposure to human beings, MG possesses a potential environmental health hazard. So, we performed with HepG2, a human hepatocellular carcinoma cell line, to identify the differentially expressed genes (DEGs) related to toxicity of MG. And we compared gene expression between control and MG treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity $(IC_{20})$ of MG was determined above the $0.867{\mu}M$ in HepG2 cell for 48 h treatment. And the DEGs of MG were identified that 5 out of 6 DEGs were upregulated and 1 out of 6 DEGs was down-regulated by MG. Also, MG induced late apoptosis and necrosis in a dose dependent in flow cytometric analysis. Through further investigation, we will identify more meaningful and useful DEGs on MG, and then can get the information on mechanism and pathway associated with toxicity of MG.

Effects of Therapeutic Massage and Home Exercise Program on Pain, Range of Motion, Depression, and Quality of Life after Arthroscopic Rotator Cuff Repair

  • Lee, Young-Min;Kwon, Jung-Won
    • The Journal of Korean Physical Therapy
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    • v.33 no.2
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    • pp.114-121
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    • 2021
  • Purpose: This study examined the effects of therapeutic massage (TMP) and home exercise program (HEP) on pain, range of motion (ROM), depression, and quality of life (QOL) after arthroscopic rotator cuff repair in middle-aged women. Methods: Nineteen middle-aged women who underwent arthroscopic rotator cuff repair were enrolled in the study. All subjects were assigned randomly to the experimental group (n=9) or the control group (n=10). The subjects in both groups performed HEP for 30 minutes, three times per day, for eight weeks. The subjects in the experimental group also performed HEP, with TMP being performed for 30 minutes, 16 sessions for eight weeks. All subjects were evaluated with a visual analogue scale (VAS), shoulder ROM, self-rating depression scale (SDS), and Korean brief version of WHO quality of life scale (WHOQOL-BREF). A paired t-test was performed to test the significant differences between before and after the intervention. An independent t-test was conducted to test the significant differences between the groups. Results: Following the intervention, both groups showed significant differences than before the intervention in the VAS, shoulder ROM, SDS, and WHOQOL-BREF (p<0.05). There were significant differences in the ROM of shoulder flexion, SDS, and WHOQOL-BREF between the groups (p<0.05). Conclusion: These findings suggest that intensive TMP is an effective method for improving shoulder flexion mobility, depressive symptom, and QOL in middle-aged women after arthroscopic rotator cuff repair.

Anticancer effect of Rheum Rhizoma on human liver cancer HepG2 cells (간암 세포주 HepG2에 대한 대황 추출물의 항암효과)

  • Yun, Hyun-Joung;Hwang, Seong-Goo;Yun, Hyung-Joong;Kim, Chang-Hyun;Seo, Gyo-Soo;Park, Won-Hwan;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.27-36
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    • 2006
  • Objectives : This study was performed for the investigation of anticancer effects of methanol extract of Rheum Rhizoma (MeOH-RR) on a human liver cancer cell line (HepG2). Methods : To study the cytotoxic effect of MeOH-RR on HepG2 cells, the cell viability was determined by XTT reduction method and trypan blue exclusion assay. The cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, MeOH-RR-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome c from mitochondria to cytosol, the level of Bcl-2 and Bax were examined by western blot analysis. Results : MeOH-RR reduced proliferation of HepG2 cells in a dose-dependent manner at 24 h and 48 h treatment. MeOH-RR induced the activation of caspase-3, -8, and -9 and the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3. Furthermore, treatment with MeOH-RR resulted in internucleosomal DNA fragmentation, evidenced by the formation of a DNA ladder on agarose gel, a hallmark of cells undergoing apoptosis. MeOH-RR downregulated Bcl-2, upregulated Bax, and increased the release of cytochrome c from the mitochondria into cytosol in a dose-dependent manner. Moreover, MeOH-RP increased caspase-3 activity. Conclusion : There results suggest that MeOH-RR induce apoptosis via mitochondrial pathway and caspase-3-dependent pathway in HepG2 cells. There results suggest that MeOH-RR is potentially useful as a chemotherapeutic agent in human liver cancer.

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Growth Inhibitory and Quinone Reductase Activity Stimulating Effects of Internal Organs of Aplysia kurodai Fractions on Cancer Cell Lines In vitro (군소내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase 유도 활성 증가 효과)

  • Shin, Mi-Ok
    • Journal of Life Science
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    • v.20 no.6
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    • pp.877-884
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    • 2010
  • We investigated the growth inhibitory effect of internal organs of Aplysia kurodai (AK) on proliferation in cancer cell lines in vitro. The internal organs of AK were extracted with methanol (AKM), which were then further fractionated into four subfractions by using solvent partition method, resulting in hexane (AKMH), methanol (AKMM), butanol (AKMB), and aqueous (AKMA) soluble fractions. We determined the cytotoxic effect of these four fractions in four kinds of cancer cell lines - HepG2, MCF-7, HT29 and B16-F10 - by MTT assay. Among the four subfractions of AKM, AKMM showed the strongest cytotoxic effects on all cancer cell lines which were used. Morphological changes such as membrane shrinking and blebbing of cells were also observed in AKMM treatment in HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol layer (AKMM) of HepG2 cells. AKMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The QR induced effect of AKMM was determined to be 2.4 at $100\;{\mu}g/ml$ level with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Aplysia kurodai (AK) may be a chemopreventive agent for the treatment of human cells.