• Title/Summary/Keyword: MTT Reduction Assay

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Cytoprotective Effect by Antioxidant Activity of Codonopsis lanceolata and Platycodon grandiflorum Ethyl Acetate Fraction in Human HepG2 Cells (인간 HepG2 세포에서 더덕 및 도라지 에틸아세테이트 분획물의 항산화 효과에 의한 세포보호 효과)

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Oh, Hyun-Taek;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.696-701
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    • 2008
  • The objective of this study was to determine whether Codonopsis lanceolata or Platycodon grandiflorum ethyl acetate fraction (CLEA or PGEA) protect cells against sodium nitroprusside (SNP)-induced oxidative stress via the expression of various antioxidant systems. The HepG2 cells exposed for 24 hr to 0.5 mM SNP showed a reduction in the cell viability by an MTT assay. Pretreatment with CLEA and PGEA resulted in an inhibition of SNP-induced cell death. In addition, the effects of CLEA and PGEA on the expression of antioxidant systems via RT-PCR analyses was assessed. The levels of catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD) and metallothionein (MT)-1A mRNA were increased after 24 hr of CLEA exposure. The levels of Mn superoxide dismutase CAT, G6PD, MT-1A, and MT-2A mRNA were increased after PGEA treatment. In conclusion, CLEA and PGEA exert indirect antioxidant effects, perhaps via the induction of a variety of antioxidant systems which, may protect cells against oxidative stress.

Bioactivity and Chemical Composition of the Essential oil of Tea Tree (Melaleuca alternifolia) (티트리 에센셜오일의 생물활성 및 주요 성분 분석)

  • Yang, Seun-Ah;Jeon, Sang-Kyung;Lee, Eun-Jung;Im, Nam-Kyung;Jung, Ji-Young;Lee, In-Seon
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1644-1650
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    • 2008
  • The essential oil of tea tree (Melaleuca alternifolia) is widely used in traditional Australian medicine for skin lesions and infected injuries. In the present study, we investigated the chemical composition, cytotoxicity and its biological activities. The composition of the oil was analyzed by GC-MS. ${\beta}$-Terpinene (20.87%), ${\alpha}$-pinene (17.60%), p-cymene (11.23%), 3-carene (10.40%), trans-anethole (8.47%) and limonene (4.65%) were the major components in the oil. The results tested by MTT assay indicated that the oil showed no cytotoxic effect, at concentrations up to 5%, for less than 3h. The antiradical capacity was evaluated by measuring the scavenging activity of the essential oil on the 2,20-diphenylpicrylhydrazyl (DPPH) and 2,2'-azino-bis 3-ethyl benzothiazoline-6-sulfonic acid (ABTS) radicals. The oil was able to reduce the both radicals dose-dependently, and the concentration required for 50% reduction ($RC_{50}$) against ABTS radicals ($1.6{\pm}0.02%$) was slightly lower than DPPH radicals ($2.6{\pm}0.29%$). The direct contact and vapor-phase antibacterial activity of the oil were also evaluated using disc diffusion method against Staphylococcus aureus, Streptococcus mutans, Listeria monocytogenes, Acinetobacter baumannii, Escherichia coli, and Vibrio parahaemolyticus. All the Gram-negative bacterial strains tested showed more sensibility to the oil than the Gram-positive strains when compare to the effect of gentamycin. On the other hand, the vapor phase of the essential oil against S. aureus exhibited strongest inhibitory effect.

Experimental Studies on the Anti-tumor and the Immunomodulatory Effects of Jiaweicitaowan(加未慈桃丸) (가미자도환(加味慈桃丸)의 항암(抗癌) 및 면역증강효과(免疫增强效果)에 관한 부험적(實驗的) 연구(硏究))

  • Jeon, Young-Soo;Shim, Bum-Sang;Choi, Seung-Hoon;Ahn, Kyoo-Sook
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.8 no.1
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    • pp.103-125
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    • 2002
  • This experimental study was carried out to evaluate the anti-tumor and the immunomodulatory effects of Jiaweicitaowan(加未慈桃丸) against cancer. The in vitro anti-tumor effects were evaluated by MTT assay. The cytotoxicity, extension of survival days, the effect of inhibition solid tumor which was induced sarcoma 180, and the changes of body weight were evaluated for in vivo effects of anti-tumor. To evaluate the immunomodulatory effects of Jiawei- citaowan(加未慈桃丸), delayed type hypersensitivity, hemagglutinin, hemolysin titers for humoral immune response, rosette forming cells for cell-mediated immune response, natural killer cell activity, proliferation of lymphocyte, productivty of Interleukin-2, and carbon clearance were measured with methotrexate treated mice. The results were as follows; 1. In the case of existence ability of tumor cell, IC50 had an anti-tumor ativity resulted 2.52mg/ml to SNU-C4. 0.41mg/ml to SNU-396, resulted to 0.09mg/mlSNU-1. 2. The groups of Jiaweicitaowan(加未慈桃丸) 10mg/ml, 20mg/kg had no body weight loss. reduction in intake of water and feed, so these had no toxicity. 3. In the case of the effect of extention of existence. the group of 20mg/kg Jiaweicitaowan(加未慈桃丸) extract treated group was showed 250% in ILS. 4. The effect of inhibition solid tumor was significantly decreased in both 10mg/kg, 20mg/kg of Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group S. The groups of 10mg/kg, 20mg/kg Jiaweicitaowan(加未慈桃丸) had significant effect of body weight change compared to control group. 6. Delayed type hypersensitivity was not significant in both Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 7. Hemagglutinin and Hemolysin titers were significantly increased by dose-dependent. so these results showed that the humoral immume respose was activated. 8. For the effect of rosette formimg cells was not significant in hoth Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 9. Natural killer cell activity was significantly increased in both Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group in the ratio of 100: 1, 50: 1 of effector and target cells, but in the ratio of 10:1, the Jiaweicitaowan(加未慈桃丸) extract treated groups were not significant. 10. The proliferation of lymphocyte and productivty of Interleukin-2 were significantly increased by dose-dependent in both 10mg/ kg, 20mg/ kg of Jiaweicitaowan(加未慈桃丸) extract treated groups as compared with control group. 11. In the phagocytic effect, the 20mg/kg of Jiaweicitaowan(加未慈桃丸) extract treated group showed the increasing effect with significance as compared with control group. According to the results, we can suggest that Jiaweicitaowan(加未慈桃丸) has the antitumor and the immunomodulatory effects.

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Effects of Ethanol Extracts from Commonly Consumed Vegetables by Koreans on NO, TNF-${\alpha}$ and MCP-1 Production in LPS-stimulated RAW 264.7 Macrophages (한국인 다소비 채소의 에탄올 추출물이 LPS 처리된 대식세포에서 NO, TNF-${\alpha}$와 MCP-1 분비에 미치는 영향)

  • Ahn, Eun Mi;Kang, Hyun Ju;Park, Young-Hee;Kim, Young;Kang, Min-Sook
    • Journal of the East Asian Society of Dietary Life
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    • v.24 no.6
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    • pp.776-784
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    • 2014
  • Korean dishes, Hansik are characterized by healthful vegetable intake. The purpose of this study was to evaluate the inhibitory effect of commonly consumed vegetables by Koreans on obesity/metabolic disease-related inflammation. Through statistical analysis of the KNHANES database ($1^{st}$ 1998, $5^{th}$ 2010, 2011) and a literature review, we selected vegetables for study. Among the vegetables, main or sub ingredients of Kimchi were excluded. Samples were prepared using only edible portions and freeze-dried. After grinding, samples were extracted with ethanol, evaporated and finally lyophilized. The cytotoxicity of samples was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, at various concentrations that do not affect cell viability. Raw 264.7 macrophages were treated with lipopolysaccharide (LPS) and 11 kinds of samples or positive control (troglitazone) dissolved in dimethyl sulfoxide (DMSO). After 24 hours, nitric oxide (NO), tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) and monocyte chemoattractant protein-1 (MCP-1) production were determined. Excepts for young pumpkin and bracken, nine samples effectively reduced NO production compared with control treated with LPS and DMSO. NO levels of five samples (bean sprouts, leeks, eggplant, mugwort, and pumpkin) were similar to that of the positive control. These five samples showed significantly decreased TNF-${\alpha}$ or MCP-1 compared to the control group. Our results suggest that consumption of commonly consumed vegetables contributes to partial prevention of obesity and related metabolic syndrome through reduction of NO, TNF-${\alpha}$, and MCP-1 production.

Anti-cancer Effects of Samultang-Gami on HeLa, HepG2, AGS Cells (사물탕가미방(四物湯加味方)의 항암효과에 대한 실험적 연구)

  • Jung, Jae-Joong;Goo, Sun-Young;Goo, Sun-Young;Sung, Jung-Suk;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.3
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    • pp.38-55
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    • 2010
  • Purpose: This study was designed to find out the anti-cancer effects of Samultang-Gami which was composed of Rehmanniae Radix(RR), Angelicae Gigantis Radix(AGR), Cnidii Rhizoma(CR), Paeoniae Radix(PR), Cortex Moutan Radicis(CMR), Hedyotis Diffusa(HD) and Caesalpinia Sappan on HeLa, HepG2 and AGS cells. Methods: Various cancer cell lines including HeLa, HepG2 and AGS cells, were used. In vitro anti-cancer effects were measured by MTT assay using cancer cell lines treated with various concentrations of 70% ethanol extract of Samultang-Gami. Expression of cell cycle arrest mediators including Bax, Bcl-2, p53 and DARP-1 proteins were measured by Western blot analysis. Results: 1. Samultang-Gami decreased the viability of HeLa and HepG cells in a dosedependent manner. 2. AGR, CMR, PR and HD decreased the viability of HeLa, HepG2 and AGS cells. 3. We could observe that the decreased Bax and Bcl-2 expression level and the increased PARP-1 expression level by Samultang-Gami extracts treated in HeLa cells. 4. We could observe that the decreased Bcl-2 expression level and the increased Bax, p53 and PARP-1 expression level by RR extracts treated in HeLa cells. and also could observe that the reduction of the protein level of Bcl-2, p53 and PARP-1 and the increase of the protein level of Bax by PR in HeLa cells. 5. We could observe that the increased p53 expression level, the decreased PARP-1's that and the unchanged Bax and Bcl-2's that by Samultang-Gami extracts treated in HepG2 cells. 6. We could observe that the reduced Bcl-2 expression level by each of RR extracts and PR extracts in HepG2 cells. 7. The treatment of Samultang-Gami in AGS cells didn't have any effect on the expression level of Bax, Bcl-2, p53 and PARP-1. 8. We could observe that the increased p53 and PARP-1 expression level by each of CR, RR and PR extracts in AGS cells. Conclusion: Taken together, we suggest that Samultang-Gami exhibits cytotoxic effects on HeLa, HepG2 and AGS cells, causing apoptosis. The results showed that Samultang-Gami may do so by regulating the expression of specific target molecules that promote efficient apoptotic cell death in a dose-dependent manner.

Beneficial effect of Orostachys japonicus A. berger herbal acupuncture on oxidant-induced cell injury in renal epithelial cell (와송약침액이 Oxidant에 의한 신장세포손상에 미치는 영향)

  • Park, Sang-Won;Kim, Cheol-Hong;Youn, Hyoun-Min;Jang, Kyung-Jeon;Ahn, Chang-Beohm;Song, Choon-Ho
    • Korean Journal of Acupuncture
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    • v.24 no.1
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    • pp.171-187
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    • 2007
  • Objectives : This study was performed to determine if Orostachys japonicus A. Berger herbal acupuncture (OjB) provides the protective effect against the loss of cell viability and DNA damage induced by oxidant in renal proximal tubular cells. Methods : The cell viability was evaluated by a MTT reduction assay and DNA damage was estimated by measuring double stranded DNA breaks in opossum kidney (OK) cells, an established proximal tubular cell line. Lipid peroxidation was determined by measuring malondialdehyde (MDA), a product of lipid peroxidation. Results : H2O2 increased the loss of cell viability in a time-dependent manner, which were prevented by 0.1% OjB. The protective effect of OjB was dose-dependent over concentration range of 0.05-0.5%. H2O2 caused ATP depletion and DNA damage, which were prevented by OjB and the hydrogen peroxide scavenger catalase. The loss of cell viability by H2O2 was not affected by the antioxidant DPPD, but lipid peroxidation by the oxidant was completely inhibited by DPPD. Generation of superoxide and H2O2 in neutrophils activated by phorbol-12,13-dibutyrate was inhibited by OjB in a dose-dependent manner. OjB inhibited generation of H2O2 in OK cells treated with antimycin A and exerted a direct H2O2 scavenging effect. Exposure of OK cells to 1 mM tBHP caused a significant depletion of glutathione which was prevented by OjB. OjB accelerated the recovery in cells cultured for 20 hr in normal medium without oxidant following oxidative stress. Conclusions : These results suggest that OjB exerts the protective effect against oxidant-induced cell injury and its protective effect was resulted from radical scavenging and antioxidant activities.

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Protective Effect of Propofol against Hypoxia-reoxygenation Injury in HaCaT Human Keratinocytes

  • Kim, Yong-Ho;Kang, Jin-Mo;Kim, In-Ryoung;Lee, Bo-Young;Yoon, Ji-Young;Kim, Cheul-Hong;Park, Bong-Soo
    • International Journal of Oral Biology
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    • v.39 no.2
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    • pp.97-105
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    • 2014
  • The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% $CO_2$, 1% $O_2$, 94% $N_2$) followed by 12 h of reoxygenation (5% $CO_2$, 21% $O_2$, 74% $N_2$). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and $100{\mu}M$ Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and $100{\mu}M$ was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

Protective effects of mulberry (Morus alba) sugar extracts on hydrogen peroxide-induced oxidative stress in HepG2 cell (오디 당침출액의 HepG2 세포에서 H2O2로 야기된 산화적 스트레스 보호 효과)

  • Youn, Young;Kim, Ha-Yan;Park, Hoe-Man;Lee, Sun-Ho;Park, Jong-Ryul;Hong, Seong-Gi;Kim, Young-Geun
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.751-757
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    • 2015
  • The objective of this study was to investigate the protective effects of mulberry (Morus alba) sugar extracts (MSE) against $H_2O_2$-induced oxidative stress in HepG2 cells. The MSEs was mixed with matured mulberry and sugar at the same ratio (1:1, w/w) and stored at $18{\pm}3^{\circ}C$ for 40 days. In 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging test, MSE stored for 40 days showed high activity with a ratio above 66%. Therefore, we selected 40 days as the optimum storage period. After cell viability analysis using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, we determined that the optimum concentration of MSE was 0.5%. Our results showed that MSE increased the cell viability and antioxidant enzyme activities of superoxide dismutase (SOD) and catalase in $H_2O_2$-treated HepG2 cells. Moreover, the treatment with MSE inhibited malondialdehyde (MDA) levels in $H_2O_2$-treated HepG2 cells. We also observed a reduction in apoptotic bodies in the Hoechst staining. These data show that MSE treatment significantly suppressed caspase-3 activity in HepG2 cells expored to $H_2O_2$-induced oxidative stress, thereby indicationg the protective effects of MSE in $H_2O_2$-induced oxidative stress.

Growth-inhibitory Effects of the Plocamium telfairiae Extracts on Cancer Cells (참곱슬이(Plocamium telfairiae) 추출물의 암세포 성장억제 효과)

  • Kim, Ju-Young;Hwang, Ji-Hwan;Cha, Mi-Ran;Choi, Byeong-Dae;Choi, Sun-Uk;Park, Hae-Ryong;Hwang, Yong-Il
    • Journal of Life Science
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    • v.16 no.4
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    • pp.659-663
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    • 2006
  • The extracts of Plocamium telfairiae using several solvents with different polarities were prepared and their growth inhibitory effects were examined on the human cancer cells. We investigated the cytotoxic effects of P. telfairiae extracts on HT-29 cells by the MTT reduction assay and examining the morphological change under the inverted microscope. Among three extracts, the methanol extract showed the strongest inhibitory effect on the growth of HT-29 cells. The methanol extract was further fractionated sequentially with n-hexane, diethyl ether, ethyl acetate, and aqueous for purifying crude methanol extract. The n-hexane layer among the fractioned layers showed remarkable inhibitory activity on the growth of HT-29 cells. Moreover n-hexane layer showed the notable growth inhibition effects with a dose-dependent manner against SW620, HeLa, and MCF-7 cells as well as HT-29 cells. These results indicated that P. telfairiae extracts may be contained bioactive materials with inhibitory effect on the growth of human cancer cells.

Effects of Jagamchotang on the Cultured Rat Neonatal Myocardial Cells (자감초탕(炙甘草湯)이 배양심근세포(培養心筋細胞)에 미치는 영향(影響))

  • Lee, Lae-Chun;Cho, Nam-Su;Cho, Dong-Ki;Eom, Sang-Sup;Kang, Sung-Do;Lee, Chun-Woo;Go, Jeong-Soo;Sung, Yeun-Kyung;Lee, Kwan-Hyung;Sung, Ki-Ho;Park, Jun-Su;Ryu, Do-Gon;Moon, Byung-Sun
    • Journal of Oriental Physiology
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    • v.14 no.2 s.20
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    • pp.179-187
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    • 1999
  • To investigate how Jagamchotang provent cellular injury by a certain starting point on reperfusion injury after ischemia in myocardial cell, conducted MTT assay, LM stydy and measured LDH secretion, heart rate and nitric oxide(NO), and got the following results. 1. Jagamchotang did not injure cells even in $20{\mu}g/ml$. 2. Jaganchotang repressed the toxicity of mitochondria and cell membrane in reperfusing after ischemia and repressed the contraction of promontory of myocardial cell and reduction of the number of cells. Also maintained regular heart rate and reduced the number of heart rate. 3. Synthesis of NO by Jagamchotang in ischemia increased 1.9 times than a control. 4. When reperfusing with sodium nitropruside (SNO), NO donor in ischemia repressed the toxicity of mitochondria as the case of reperfusing with Jagamchotang in ischemia. Therefore, putting these findings together, it. can be said the effect of Jagamchotang in ischemia will be closely related with generation of NO.

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