• Title/Summary/Keyword: Cytotoxicity

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Anti-Cancer Effects of Peptides Purified from Culture Supernatant of Lactobacillus casei (Lactobacillus casei의 배양물에서 분리한 물질의 항암 효과)

  • Kim, Jung-Hwa;Kim, Dong-Myung;Baek, Hong;Lee, Seung-Hoon;Chung, Myung-June
    • Journal of Dairy Science and Biotechnology
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    • v.26 no.1
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    • pp.5-10
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    • 2008
  • This study was conducted to isolate protein components from culture supernatant of Lactobacillus casei. and measure anti-cancer activity. The protein components were isolated A and B on Ultrafiltration membrane(3, 10, 30, 100 KDa). And the protein components A and B were isolated fractions(number $3{\sim}9$) on FPLC. Experimental studies were progressed through the cell cytotoxicity and anti-cancer activities. Cell cytotoxicity test using human kidney normal cell(293) showed cytotoxicity of below 20% by the protein components A and B($100{\mu}g/mL$). The anti-cancer activity was increased up to 70% by the protein components A and B($100{\mu}g/mL$) in AGS(stomach cancer), A549(lung cancer), MCF-7 (breast cancer), SK-OV-3(ovary cancer) and LoVo(colon cancer). Cell cytotoxicity test was showed cytotoxicity of about 50% by the fractions(number 3, 8, 9) isolated FPLC. The others have not the cytotoxicity about the human normal cell. The anti-cancer activity was increased up to 70% by the fraction number 7 in cancer cell line. Therefore the components isolated from culture supernatant of Lactobacillus casei were showed anti-cancer activity.

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Antioxidative and Hepatoprotective Effects of Injinho-Tang and Osumogwa-Tang (인진호탕(茵蔯蒿湯)과 오수목과탕(吳茱木瓜湯)의 항산화(抗酸化) 및 간세포(肝細胞) 보호효과(保護效果))

  • Lee, Sang-Hyun;Kim, Young-Bok
    • Herbal Formula Science
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    • v.16 no.1
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    • pp.117-130
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    • 2008
  • Korean traditional medicine has been used for the treatment of the various diseases based on both oriental medicinal theory and clinical trials. Thus, the prescriptions of Korean traditional medicine would be useful for the development of new therapeutics. This research focuses on the fundamental study in Korean traditional prescriptions for the development of new hepatoprotective agents. We found two prescriptions. Injinho-Tang and Osumogwa-Tang, showed the significant DPPH free radical scavenging and hepatoprotective effect, respectively. It is well-known that free radical scavenging effect is related to the prevention of various pathological events including liver injury. This paper deals with hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, free radicals scavenging on both DPPH and superoxide of above two prescriptions. Hot water extract of Injinho-Tang did not show the significant hepatoprotective effect on tacrine-induced cytotoxicity in Hep G2 cells, however, it shows the significant scavenging effects for both DPPH and superoxide radicals. On the other hand, all of the hot water extracts of constituent herbal drugs in Injinho-Tang exhibited the promising protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Of these, water extract of Rhei Rhizoma showed the most prominent effect on tacrine-induced cytotoxicity in Hep G2 cells. Bioassay-guided fractionation of Rhei Rhizoma extract has furnished four compounds, and their chemical structures have been identified by comparison of their spectral data with those of literature as chrysophanol (1), emodin (2), 3,5-dihydroxy-4'- methoxystilbene (3), and rhapontigenin (4), respectively. Among the isolated compounds, compounds 2-4 revealed the significant hepatoprotective effect in vitro when their $EC_{50}$ values compare with that of silybin, as a positive control. It also exhibited that emodin possessed the most hepatoprotective effect among these active compounds. In case of Osumogwa-Tang, its hot water extract showed the moderate protective effect on tacrine-induced cytotoxicity in Hep G2 cells. Hot water extract of Chaenomelis Fructus, one of the constituent herbal drug of this prescription, exhibited the significant hepatoprotective effect with $EC_{50}$ value of $7.8{\pm}0.1\;{\mu}g/ml$, however, it showed strong cytotoxicity in Hep G2 cells above the concentration of $25\;{\mu}g/ml$. It was revealed that both hot water extract of Evodiae Fructus and its butanol soluble fraction showed the moderate hepatoprotective effect but concentration-dependent activity in Hep G2 assay system. Two quinolone alkaloids, evocarpine and dihydroevocarpine, also tested for their hepatoprotective effects on tacrine-induced cytotoxicity in Hep G2 cells, however, these two compounds derived from the Evodiae Fructus as the major constituents did not show in vitro hepatoprotective effect. From these results, it would be necessary to further isolation of its hepatoprotective compounds from the butanol soluble fraction of the hot water extract of Evodiae Fructus.

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In vitro Cytotoxicity Evaluation of Polydimethylsiloxane as a Biosensor Coating Material (바이오센서 코팅용 Polydimethylsiloxane의 생체외 세포독성 평가)

  • Park, Subeom;Lee, Jonghwan;Na, Kyunga;Jung, Jaeyeon;Kim, Myungjin;Park, Sungjae;Hyun, Jinho
    • Journal of Adhesion and Interface
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    • v.10 no.2
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    • pp.77-83
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    • 2009
  • PDMS was selected for a coating material of implantable biosensors and the cytotoxicity of extracts released from a polymer was evaluated using ISO 10993-5, Biological evaluation of medical devices-Part 5: Tests for in vitro cytotoxicity. Organo-tin was used as a positive control and a medium without serum was used as a negative control. Materials extract were prepared by incubating specimens in RPMI medium without serum ($125{\mu}L/cm^2$) for 24 h, 1 week and 6 weeks at $38^{\circ}C$. The evaluation of cytotoxicity was performed by two different methods : 1) seeding cells with extracts at the beginning 2) incubating extracts with cell sheets already formed on the plate. Both cell morphology and MTT numerical data were shown for the confirmation of cytotoxicity and cell spreading on the surface of PDMS.

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Antioxidative Effect of Chelidonium majus Extract on Cultured NIH3T3 Fibroblasts Injured by Cadmium Chloride of Toxicant (독성물질인 염화카드뮴으로 손상된 배양 NIH3T3 섬유모세포에 대한 애기똥풀 추출물의 항산화 효과)

  • Kim, Tae-Yoon;Jekal, Seung-Joo
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.1
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    • pp.1-7
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    • 2016
  • The aim of this study was to evaluate the cytotoxicity of cadmium chloride ($CdCl_2$), toxicant, and the protective effect of Chelidonium majus (CM) extract on $CdCl_2$-induced cytotoxicity in cultured NIH3T3 fibroblasts. Cell viability, the effect of butylated hydroxytoluene (BHT) against $CdCl_2$, and the antioxidative effects including DPPH-free radical scavenging activity, superoxide anion-radical scavenging activity (SSA), and lactate dehydrogenase (LDH) activity were assessed. $CdCl_2$ caused a significant dose-dependent decrease in cell viability, and $XTT_{50}$ value was determined at 38.7uM of $CdCl_2$. It was determined as highly-toxic by Borenfreund and Puerner' toxic criteria. BHT of antioxidant significantly increased cell viability severely damaged by $CdCl_2$-induced cytotoxicity in these cultures. In the protective effect of CM extract on $CdCl_2$-induced cytotoxicity, CM extract significantly increased cell viability, DPPH-free radical scavenging activity, SSA and inhibitory activity of LDH. From these results, it is suggested that oxidative stress is involved in the cytotoxicity of $CdCl_2$, and CM extract showed protective efficacy on $CdCl_2$-induced cytotoxicity via antioxidative effects. Conclusively, natural resources like CM extract may be a putative antioxidative agent for the detoxification or diminution of toxicity correlated with oxidative stress.

Effect of Smilax China L. Extract on Cultured NIH3T3 Fibroblasts Damaged by Mercury as Allergic Contact Dermatitis Inducer (알러지성 접촉피부염 유발제인 수은으로 손상된 배양 NIH3T3 섬유모세포에 대한 청미래덩굴 추출물의 효과)

  • Han, Sun-Hee;Jekal, Seung-Joo
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.175-181
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    • 2015
  • In order to examine the effect of Smilax china L. (SC) extract on the cytotoxicity of methymercuric chloride (MMC), allergic contact dermatitis, The cytotoxicity of MMC was assessed after cultured NIH3T3 fibroblasts were treated with various concentrations of MMC for 72 hours. And also, the following results were obtained by measuring the antioxidative effect of SC extract on the cytotoxicity of MMC. In this study, MMC remarkably decreased the cell viability of NIH3T3 fibroblasts in a dose-dependent manner, and MMC was seen to be highly-toxic below 100 uM of $XTT_{50}$ value. In addition, the toxicity of MMC was involved in oxidative stress via a blockage of MMC-induced cytotoxicity by vit. E as antioxidant. In the protective effect of SC extract on MMC-induced cytotoxicity, SC extract defended the cytotoxicity of MMC by a significant increase of cell viability which was decreased by MMC-induced cytotoxicity. It also showed antioxidative effects such as electron donating ability (EDA), superoxide dismutase (SOD)-like activity (SLA) and the lipid peroxidation activity (LPA). From these results, the natural component as SC extract may be a putative resource as the antioxidative agent for the treatment of inflammatory skin disease associated with the oxidative stress.

Study on Synergistic Anti-tumor Effect of Combination with Adriamycin and Palginhonhapwhajucwhan (팔진탕합화적환(八珍湯合化積丸)과 Adriamycin의 병용처리시 나타나는 synergistic 항종양(抗腫瘍) 효과(效果)에 관(關)한 작용기전 연구(硏究))

  • Moon, Gu;Moon, Seok-Jae;Won, Jin-Hee;Cho, Jung-Yun;Park, Sang-Gu;Song, Bong-Gil;Park, Rae-Gil;Lee, Byung-Gu
    • The Journal of Internal Korean Medicine
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    • v.21 no.3
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    • pp.443-452
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    • 2000
  • Objective : This study was designed to evaluate the synergistic effect on cytotoxicity of combination with adriamycin and Palginhonhapwhajucwhan, a traditional prescription for cancer treatment in oriental medicine, in Chang, HL-60, Hep-3B and Alexander cells. Methods : We observed cell viability in Chang, HL-60, Hep-3B, and Alexander cells by crystal violet staining. Those cells were treated with various concentrations of adriamycin alone, Palginhonhapwhajucwhan alone and combination of two medications for 10 hr. On condition of $0.5{\mu}l/ml$ adriamycin alone, $15.6{\mu}l/ml$ Paljintanghapwhajucwhan alone and combination of two medications, at first, we observed colony forming of Chang and HL-60 cells. Second, we observed DNA fragmentation by agarose electrophoresis in Chang, HL-60, Hep-38 and Alexander cells. Third, we measured the catalytic activation of caspase-1, 2, 3, 6, 8, and 9 protease in Chang cells and caspase-3 protease in Chang, HL-60, Hep-3B and Alexander cells by using fluorogenic substrate. Finally, we isolated mRNA of Fas in Chang, HL-60, Hep-38 and Alexander cells and observed that Fas gene was amplified by RT-PCR Results : 1. The combination of adriamycin and Palginhonhapwhajucwhan synergistically augmented the cytotoxicity of Chang and HL-60 cells whereas did not in Hep-38 and Alexander cells. 2. Cotreatment of two drugs also markedly inhibited the colony forming ability both in Chang and HL-60 cells. 3. The cytotoxicity of these medicatons was revealed as apoptosis characterized by high molecular wight DNA fragmentaton. 4. The apoptotic cytotoxicity was mediated by activation of caspase-3 protease in Chang cells. 5. Synergistic increase in apoptotic cytotoxicity by combination of two medications was dependent on the expression of Fas in cancer cells. Conclusions : Combination of adriamycin and Palginhonhapwhajucwhan significantly augmented apoptotic cytotoxicity of Fas-positive cells such as Chang and HL-60 cells via acticaton of apoptosis signaling pathway.

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The effects of antibodies and complement in macrophage-mediated cytotoxicity on metacercariae of the lung fluke, Paragonimus westeymani (폐흡충(Paragonimus Tuestermani) 피낭유충에 대한 대식세포의 세포독성에 있어서 항체 및 보체가 미치는 영향)

  • 민득영;안명희
    • Parasites, Hosts and Diseases
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    • v.28 no.2
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    • pp.91-100
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    • 1990
  • Paragonimus westermani is a tissue migrating parasite in the early stage until arriving at lung, and most of the parasites spend their life spans there. Considerable immune responses including activation of macrophages are taken place during the residence of parasites in the host. However, concerning the immunologic defense mechanisms of the host against this parasite, only a few document is available so far. In this study, the cytotoxic effect of peritoneal macrophages under the presence of antibody and/or complement against metacercariae of F. westermani was investigated in vitro. Metacercarlae were collected from the crayfish, Cambaroides similis and hatched out in Tyrode solution (pH 7.4). Plastic adherent cells from normal or infected rat (Wistar) peritoneal exudates were used as experimental macrophages. Polyclonal antibodies were obtained from infected rats and a cat. Cat IgG was fractioned with ion exchange chromatography. Fresh rabbit complement was used according to experimental scheme. Various combinations of peritoneal macrophages, normal or infected rat serum, complement and cat IgG were incubated at $36^{\circ}C$ in 5% $CO_2$ incubator for 6, 14, 24 and 48 hours. The results obtained were as follows: 1. P. westermani infection activated peritoneal macrophages non-specifically and this activation induced increases of cell adherence and cytotoxicity on metacercariae. 2. In the presence of infected rat serum the antibody.dependent cell-mediated cytotoxicity of peritoneal macrophages on metacercariae was significantly increased and showed a peak at 6-hour incubation. But the cytotoxic effect was markedly reduced after inactivation of complement and heat.labile IgE antibody by the heating of infected serum at 56$^{\circ}C$ for 30 minutes. 3. The highest cytotoxic effect (100%) of concomitant incubation with IgG and complement showed 24 hours after incubation, although cell adherence was relatively low at 6-hour incubation and 0% at 24-hour incubation. 4. Coordinative functions of complement with serum and IgG were effective in cell adherence and in cytotoxicity, but it is not clear the independent role of complement on the macrophage- mediated cytotoxicity in this study- With these results it is assumed that P. westermani infection can induce the non-specific activation of peritoneal macrophages, and strum antibodies including IgE antibody might enhance the cytotoxicity by macrophages,

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