• Title/Summary/Keyword: cytotoxicity assay

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Antioxidative and Anti-inflammatory Activities of Ardisia arborescens Ethanol Extract (Ardisia arborescens 에탄올 추출물의 항산화 및 항염증 활성)

  • Jin, Kyong-Suk;Lee, Ji Young;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.24 no.7
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    • pp.713-720
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    • 2014
  • In this study, the antioxidative and anti-inflammatory activities of Ardisia arborescens ethanol extract (AAEE) were evaluated using in vitro assays and a cell culture model system. AAEE exhibited potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar to ascorbic acid, which was used as a positive control. Moreover, AAEE effectively suppressed lipopolysaccharide (LPS)- and hydrogen peroxide (H2O2)-induced reactive oxygen species (ROS) in RAW 264.7 cells. Furthermore, AAEE induced the expression of antioxidative enzymes, heme oxygenase 1 (HO-1), and thioredoxin reductase 1 (TrxR1), in addition to their upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), in a dose-dependent manner. The upstream signaling pathways of mitogen-activated protein kinases (MAPKs) might regulate the modulation of HO-1, TrxR1, and Nrf2 expression. On the other hand, AAEE inhibited LPS-induced nitric oxide (NO) formation, without cytotoxicity. Suppression of NO formation was the result of AEEE-induced down-regulation of inducible NO synthase (iNOS). The suppression of NO and iNOS by AAEE might be modulated by their upstream transcription factor, nuclear factor (NF)-κB, and activator protein (AP)-1 pathways. Taken together, these results provide important new insights into the antioxidative and anti-inflammatory activities of A. arborescens. AAAEE might represent a promising material in the field of nutraceuticals.

Protective Effect against Oxidative Stress in Medicinal Plant Extracts (약용식물 추출물의 산화적 스트레스에 대한 방어효과)

  • Kim, Jeong-Hee;Lee, Eun-Ju;Shin, Dong-O;Hong, Sung-Eun;Kim, Jin-Kyu
    • Journal of Radiation Protection and Research
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    • v.25 no.1
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    • pp.37-43
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    • 2000
  • Protective effect of medicinal plant extracts against oxidative stress were screened in this study. Methanol extracts from 48 medicinal plants, which were reported to have antioxidative or anti-inflammtory effect were prepared and screened for their protective activity against chemically-Induced and radiation-induced oxidative stress by using MTT assay. Thirty three samples showed protective activity against chemically-induced oxidative stress in various extent Among those samples, extract of Glycyrrhiza uralensis revealed the strongest activity (25.9% at 100μg/m) with relatively lower cytotoxicity. Seven other samples showed higher than 20% protection at 100\mgg/m. These samples were tested for protection activity against radiation-induced oxidative stress. Methanol extract of Alpina officinarum show the highest activity (17.8% at 20μg/m). Five fractions were prepared from the each 10 methanol extracts which showed high protective activity against oxidative stress. Among those fraction samples butanol fractions of Areca catechu var. dulcissima and Spirodela polyrrhiza showed the highest protective activities (78.8% and 77.2%, respectively, at 20μg/m).

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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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Antioxidant and Whitening Activity of Essential Oils (Essential Oils의 항산화 활성 및 미백작용)

  • Lim, Hye-Won;Kim, Ju-Yeon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.3 s.52
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    • pp.265-271
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    • 2005
  • This study is performed to investigate the effects of citrus essential oils on melanin production in B16 melanoma cells and reactive oxygen species (ROS) generation in RBL 2H3 cells. Five kinds of citrus essential oil (bergamot, grapefruit, lemmon, mandarin, petigrain) did not have any influence on DPPH radical scavenger activity, cell growth and cytotoxicity in B16 melanoma cells. In purified tyrosinase assay, both mandarin and petigrain essential oils dose-dependently inhibited its activity, but bergamot did not. In 1μMαMSHstimulated B16 melanoma cells, all of 5 citrus essential oils inhibited melanin production in a_ dose dependent manner. On the other hand, four kinds of citrus essential oil dose-dependently increased ROS generation in RBL 2H3 mast cells, but mandarin did not. From the above results, it is possible that citrus essential oils nay be developed to be anti-melanogenic agent on the basis of their inhibitory effect on MSH-induced melanin production. Hut we can not rule out the possibility of the induction of allergy and inflammation since citrus essential oils caused ROS generation in RBL 2H3 mast cells.

Studies on Skin Whitening Effects of Resorcinol Dipentyl Ether (레조시놀다이펜틸에터의 피부 미백 효능 평가)

  • Cha, Young Kwon;Cho, Hyun Dae;Cho, Wan Goo;Byun, Sang Yo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.115-124
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    • 2017
  • The objective of this study was to investigate skin whitening effect of Resorcinol dipentyl ether [1,3-di(pentyloxyl)benzene] by in vitro experiments. Resorcinol dipentyl ether was prepared by alkylation of resorcinol with 1-bromopentane. The reaction products were confirmed by NMR, MS and other analytical equipments. In order to evaluate the skin safety of resorcinol dipentyl ether, the cytotoxicity of the cells constituting the skin (keratinocyte, melanocyte, fibroblast) was analyzed and similar cell viability was observed in all cell lines as compared with the control group. Inhibition of extracellular melanin synthesis effect of resorcinol dipentyl ether was approximately 65.75% at 20μg/mL and inhibition of intracellular melanin synthesis effect of resorcinol dipentyl ether was approximately 53.89% at 20μg/mL. The real-time PCR and western blot analysis of mRNA expression and protein expression of tyrosinase, TRP-1, and TRP-2 related to melanogenesis revealed that melanin inhibitory effect of resorcinol dipentyl ether was inhibited from the transcription stage respectively. Finally, this study suggested applicability of Resorcinol dipentyl ether [1,3-di(pentyloxyl)benzene] as a whitening functional cosmetic new material.

The Red-ginseng Extract Alters the Cell Cycle and Viability in the Human Neuronal Stem Cells (홍삼추출액의 인간성체신경줄기세포 증식과 세포사 관련 세포주기의 변화에 대한 효과)

  • Kim, Hyun-Jung;Kang, La-Mi;Ahn, Jin-Young;Han, Jung-Soon;Kim, Seung-U.;Lee, Kwang-Woo;Kim, Man-Ho
    • Journal of Ginseng Research
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    • v.28 no.1
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    • pp.39-44
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    • 2004
  • The present study is to determine whether the Red-ginseng extract has a proliferative or cytotoxic effect on the human neuronal stem cells(hNSCs). The hNSCs were grown and incubated with different doses of Red-ginseng extract. We tested the proliferative or cytotoxic effects by MTT and FACS analysis. Cell viability cell cycle analysis, DNA fragmentation, and bax or PARP expressions were evaluated. The hNSCs showed a proliferafe trend with its peak concentration at 0.3 μg/m. Beyond this point, higher doses decreased viabilities and showed a cytotoxic effect at 10 μg/m. There was a tendency of increased S and G2/M phases during cell proliferation. In a cytotoxic condition, decreased S phase and increased G0/G1 phases were noted, suggesting cell cycle arrest. The cytotoxic effect was associated with increase DNA fragmentation in a dose-dependent manner, However PARP cleavage or bax expression was not detected. Our results suggest that Red-ginseng extract has dual effects, the cell proliferative or cytotoxic effect, on hNSCs in vitro with dose-dependent manner.

Antioxidant and anticancer properties of hot water and ethanol extracts from the roots of Smilax china L. (발계(Smilax china L.) 열수 및 에탄올 추출물의 항산화 및 항암활성)

  • Kim, Ye Jin;Son, Dae-Yeul
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.691-698
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    • 2013
  • The biological activities of Smilax china L. rhizome (SCR), hot water (SCRW) and 70% ethanol extract (SCRE) were analyzed. The total phenolic contents of SCRW and SCRE were 51.7 and 100.5 mg/g, respectively. The measured flavonoid content of SCRW (67.7μg/g) was almost double that of SCRE (31.7μg/g). SCRE (IC50=42.4μg/mL) exhibited stronger antioxidant activity in the DPPH system than the positive control α-tocopherol (71.3μg/mL) or butylated hydroxy anisole (53.8μg/mL) did. SCRE (IC50=50.3μg/mL) also showed stronger ABTS radical scavenging activity, as did α-tocopherol (67.1μg/mL). The SOD-like activity and Tyrosinase inhibition activity of SCRW and SCRE showed almost the same pattern. The best SOD-like activity and tyrosinase inhibition activity were measured as 24.9% and 20.3% in SCRW at 1,000μg/mL, respectively. The cytotoxic effects of the SCR extracts were analyzed via MTT assay on human cancer and normal cells. SCRW and SCRE did not show cytotoxicity up to the concentration of 1,000μg/mL against the normal human cell line HEK293. Against human breast cancer cells (MCF-7), SCRW inhibited MCF-7 growth (by 27.6%) better than the anticancer drug cyclophosphamide (15.5%) at 1,000μg/mL. SCRE (1,000μg/mL) inhibited the growth of human lung cancer cells A549 (37.6%) and human stomach cancer cells AGS (53.6%) more effective than did SCRW (21.0% and 35.4%) or CPA (22.2% and 31.7%). These results suggest the potential use of SCRE and SCRW as an excellent antioxidant and antiproliferative substance, respectively.

Effects of Ethanol Extracts from Commonly Consumed Vegetables by Koreans on NO, TNF-α and MCP-1 Production in LPS-stimulated RAW 264.7 Macrophages (한국인 다소비 채소의 에탄올 추출물이 LPS 처리된 대식세포에서 NO, TNF-α와 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 (1st 1998, 5th 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 α (TNF-α) 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-α 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-α, and MCP-1 production.

Heavy Metal Ion Detection in Living Cell Using Fluorescent Chemosensor (형광화학센서를 이용한 살아있는 세포 내에서의 중금속이온검출)

  • Kwon, Pil-Seung;Kim, Jin-Kyung;Kim, Jong-Wan
    • Journal of the Korean Chemical Society
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    • v.54 no.4
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    • pp.451-459
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    • 2010
  • The fluorescence detection of intracellular metal ions are high interest in the fields of organic molecular chemistry and cellular biology. This study was purposed to detection for mercury and zinc in the cell using fluorescent chemosensor (FS). FS exhibits a weak fluorescence, but emits strong fluorescence upon Zn2+ complexation. The increased fluorescence of the 2FS/Zn2+ can be quenched completely by addition of only 1 equiv of Hg2+ with the formation of complex FS-Hg2+. Four cell lines (LLC-MK2, Hela, HT29 and AMC-HN3) were used for fluorescence imaging by confocal microscope. The cell viability MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was evaluated after cell treatment of FS, Zn2+, FS-Zn2+, Hg2+ on LLC-MK2 cell line. The cytotoxicity of FS was showed to viability over 80%. This study has shown that FS can be detected for selective imaging of Zn2+ and Hg2+ in living cells.

Ginsenoside compound-Mc1 attenuates oxidative stress and apoptosis in cardiomyocytes through an AMP-activated protein kinase-dependent mechanism

  • Hong, So-hyeon;Hwang, Hwan-Jin;Kim, Joo Won;Kim, Jung A.;Lee, You Bin;Roh, Eun;Choi, Kyung Mook;Baik, Sei Hyun;Yoo, Hye Jin
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.664-671
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    • 2020
  • Background: Ginsenoside compound-Mc1 (Mc1) is a member of the deglycosylated ginsenosides obtained from ginseng extract. Although several ginsenosides have a cardioprotective effect, this has not been demonstrated in ginsenoside Mc1. Methods: We treated H9c2 cells with hydrogen peroxide (H2O2) and ginsenoside Mc1 to evaluate the antioxidant effects of Mc1. The levels of antioxidant molecules, catalase, and superoxide dismutase 2 (SOD2) were measured, and cell viability was determined using the Bcl2-associated X protein (Bax):B-cell lymphoma-extra large ratio, a cytotoxicity assay, and flow cytometry. We generated mice with high-fat diet (HFD)-induced obesity using ginsenoside Mc1 and assessed their heart tissues to evaluate the antioxidant effect and the fibrosis-reducing capability of ginsenoside Mc1. Results: Ginsenoside Mc1 significantly increased the level of phosphorylated AMP-activated protein kinase (AMPK) in the H9c2 cells. The expression levels of catalase and SOD2 increased significantly after treatment with ginsenoside Mc1, resulting in a decrease in the production of H2O2-mediated reactive oxygen species. Treatment with ginsenoside Mc1 also significantly reduced the H2O2-mediated elevation of the Bax:Bcl2 ratio and the number of DNA-damaged cells, which was significantly attenuated by treatment with an AMPK inhibitor. Consistent with the in vitro data, ginsenoside Mc1 upregulated the levels of catalase and SOD2 and decreased the Bax:B-cell lymphoma-extra large ratio and caspase-3 activity in the heart tissues of HFD-induced obese mice, resulting in reduced collagen deposition. Conclusion: Ginsenoside Mc1 decreases oxidative stress and increases cell viability in H9c2 cells and the heart tissue isolated from HFD-fed mice via an AMPK-dependent mechanism, suggesting its potential as a novel therapeutic agent for oxidative stress-related cardiac diseases.