• Title/Summary/Keyword: In vitro cytotoxicity

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SKT + EKE (2:1) protects oxidative stress induced-liver damage (산화적 스트레스에 대한 생간건비탕가음양곽(生肝健脾湯加淫羊藿) (2:1)의 간보호효과)

  • Sang Mi Park;Dae Hwa Jung;Hyo Jeong Jin;Ye Lim Kim;Kyung-soon Kim;Min Hwangbo;Sang Chan Kim
    • Herbal Formula Science
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    • v.32 no.1
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    • pp.63-82
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    • 2024
  • Objective : Saengkankunbi-tang (SKT) is used as a traditional Korean herbal formula for treatment of liver diseases. We investigated the hepatoprotective effects of SKT plus Epimedium koreanum Nakai (EKE) against arachidonic acid (AA) + iron-mediated cytotoxicity in HepG2 cells and carbon tetrachloride (CCl4)-mediated acute liver damage in mice. Methods : The cyto-protective effects of SKT + EKE were determined by MTT assay, western blot and fluorescence activated cell sorting analysis. In mice, blood biochemistry and western blot were assessed in CCl4-induced acute hepatic damage. The animal groups included vehicle-treated control, CCl4, SKT (200 mg/kg/day), EKE (100 mg/kg/day), SKT (200 mg/kg/day) + EKE (100 mg/kg/day) and silymarin (200 mg/kg/day). Results : In HepG2 cells, pretreatment with SKT + EKE significantly suppressed cytotoxicity induced by AA + iron and reduced the expression of proteins related to apoptosis. In addition, pretreatment with SKT + EKE significantly prevented the increase in H2O2 production, GSH depletion, and lower mitochondrial membrane potential induced by AA + iron. In CCl4-induced liver damage mice, the administration of SKT + EKE prevented the liver damage by inhibition of hepatocyte damage and expression of apoptosis proteins in liver. More importantly, in vitro and in vivo assay, SKT + EKE showed significant effect compare with SKT alone or EKE alone in all parameters. Conclusions : These results indicated that SKT + EKE could protect against oxidative stress-induced liver damage, and SKT + EKE is more effective than SKT alone or EKE alone.

A COMPARISON OF RELEASE RATE AND CUMULATIVE RELEASE OF TEGDMA WITH OR WITHOUT THE APPLICATION OF BONDING RESIN (접착레진 적용 유무에 따른 TEGDMA의 방출속도 및 방출량 비교)

  • Shin, Hee-Jung;Jeon, Seong-Min
    • Restorative Dentistry and Endodontics
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    • v.23 no.2
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    • pp.701-709
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    • 1998
  • Many dental composites are Bis-GMA based resin which diluted with the more fluid monomer triethylenglycol dimethacrylate(TEGDMA). TEGDMA is often present in exess so that some quantity remains unreacted following photo-initiated polymerization. TEGDMA is a component of some resin composites which contributes to their cytotoxicity. The presence of dentin between resin composite and pulp space reduce the cytotoxicity in vitro. The root system from extrcted human third molar was removed and then a circular occlusal cavity 4mm in diameter was prepared, leaving a remaining dentinal thickness to the roof of the pulpal chamber within the range 1.0-1.5mm. Dentine was treated with 37% phosphoric acid prior to Z 100 placement without using bonding resin(group 1). In group 2, SMP(Scotchbond Multi Purpose) primer, bonding resin prior to Z 100 placement were applied sequently. In group 3, moulds with internal dimensions 4mm diameter by 2mm depth were used to contain the composite alone with an equvalent mass on tooth model, and then they were immersed directly into water. The purpose of this study is to evaluate the release rate and quantity of TEGDMA with or without the application of bonding resin. Both release rate and total cumulative amount of TEGDMA for the three groups were determined using reversed-phase HPLC at times up to 10 days. The results were as follows: 1. All experimental groups showed the highest rate of release was in the first sample period(0-4.32 min) and the rate of release declined exponentially thereafter. 2. The maximum release rate and total cumulative account of TEGDMA in the tooth model of group 1 and group 2 with the use of SMP bonding resin were reduced however ther were no significant differences between these groups(P>0.05). 3. In the first sample period(0-4.32 min), the rate of release of TEGDMA from composite resin in group 3 immersed directly into water was significantly higher than that in group 1 and group 2 of tooth model(P<0.05). Conclusively, TEGDMA diffusion from Z 100 resin was not effectively prevented by the presence of dentin in spite of using the SMP bonding resin.

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Immunotoxicological Effects of Aripiprazole: In vivo and In vitro Studies

  • Baek, Kwang-Soo;Ahn, Shinbyoung;Lee, Jaehwi;Kim, Ji Hye;Kim, Han Gyung;Kim, Eunji;Kim, Jun Ho;Sung, Nak Yoon;Yang, Sungjae;Kim, Mi Seon;Hong, Sungyoul;Kim, Jong-Hoon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.4
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    • pp.365-372
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    • 2015
  • Aripiprazole (ARI) is a commonly prescribed medication used to treat schizophrenia and bipolar disorder. To date, there have been no studies regarding the molecular pathological and immunotoxicological profiling of aripiprazole. Thus, in the present study, we prepared two different formulas of aripiprazole [Free base crystal of aripiprazole (ARPGCB) and cocrystal of aripiprazole (GCB3004)], and explored their effects on the patterns of survival and apoptosis-regulatory proteins under acute toxicity and cytotoxicity test conditions. Furthermore, we also evaluated the modulatory activity of the different formulations on the immunological responses in macrophages primed by various stimulators such as lipopolysaccharide (LPS), pam3CSK, and poly(I:C) via toll-like receptor 4 (TLR4), TLR2, and TLR3 pathways, respectively. In liver, both ARPGCB and GCB3004 produced similar toxicity profiles. In particular, these two formulas exhibited similar phospho-protein profiling of p65/nuclear factor $(NF)-{\kappa}B$, c-Jun/activator protein (AP)-1, ERK, JNK, p38, caspase 3, and bcl-2 in brain. In contrast, the patterns of these phospho-proteins were variable in other tissues. Moreover, these two formulas did not exhibit any cytotoxicity in C6 glioma cells. Finally, the two formulations at available in vivo concentrations did not block nitric oxide (NO) production from activated macrophage-like RAW264.7 cells stimulated with LPS, pam3CSK, or poly(I:C), nor did they alter the morphological changes of the activated macrophages. Taken together, our present work, as a comparative study of two different formulas of aripiprazole, suggests that these two formulas can be used to achieve similar functional activation of brain proteins related to cell survival and apoptosis and immunotoxicological activities of macrophages.

Cytotoxicity and Antitumor Effects of Insambaekhaptang on C57BL/6 Mice Melanoma-induced Lung Metastasis (인삼백합탕(人蔘百合湯)이 B16세포에 대한 세포독성능 및 C57BL/6계 생쥐의 폐전이암의 억제에 미치는 영향)

  • Hwang, Ho-Jun;Ha, Ji-Yong
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.85-98
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    • 1997
  • Oriental medicine as a candidate for effective cancer treatment recently gain positive concerns in fields of therapeutic oncology. that is why some herbal medicines have been empirically safer in toxicity than anticancer drugs used in western medicine, and to show excellent therapeutic efficacy in human trial. Thus, these effects by clinically applied-herbs have not yet fully demonstrated in experimental tumor model. This study was initiated to evaluate the antitumor effect of Insambaekhaptang as candidate of antitumor-herbal agent against B16 melanoma metastasized into C57BL/6 mice lung. In experiment to test whether Insambaekhaptang can directly kill cancer cells in vitro or not, Insambaekhaptang showed direct killing action in concentration or higher against B16 melanoma cells using MTT assay, and showed lower IC50. Another experiment to know whether Insambaekhaptang can inhibit growth and metastasis of cancer cell or not, Insambaekhaptang significantly inhibited Solid tumor by intraperiperal injected-melanoma and lung metastasis induced by intravenous injected-melanoma in inbred C57BL/6 mice. When quantitative survival time increasing, we could obtain results that increased 113% in treated by Insambaekhaptang. These results show that Insambaekhaptang can inhibit growth of B16 melanoma cells through various biological mechanisms.

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HOCl Oxidation-modified CT26 Cell Vaccine Inhibits Colon Tumor Growth in a Mouse Model

  • Zhou, Rui;Huang, Wen-Jun;Ma, Cong;Zhou, Yan;Yao, Yu-Qin;Wang, Yu-Xi;Gou, Lan-Tu;Yi, Chen;Yang, Jin-Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4037-4043
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    • 2012
  • Despite progress in elucidating mechanisms associated with colorectal cancer and improvement of treatment methods, it remains a frequent cause of death worldwide. New and more effective therapies are therefore urgently needed. Recent studies have shown that immunogenicity of whole ovarian tumor cells and subsequent T cell response were potentiated by oxidation modification with hypochlorous acid (HOCl) in vitro and ex vivo. These results prompted us to investigate the protective antitumor response with an HOCl treated CT26 colorectal cancer cell vaccine in an in vivo mouse model. Administration of HOCl modified vaccine triggered robust antitumor immunity to autologous tumor cells in mice and prolonged survival period significantly. In addition, increased necrosis and apoptosis were found in tumor tissue from the oxidation group. Interestingly, ELISPOT assays showed that specific T cell responses were not elicited in response to the immunizing cellular antigen, in contrast to raising sera antibody titer and antibody binding activity shown by ELISA assay and flow cytometry. Further evaluation of the mechanisms underlying HOCl modified vaccine mediated humoral immunity highlighted the role of antibody-dependent cell-mediated cytotoxicity. These results combined with previous studies suggest that HOCl oxidation modified whole cell vaccine has wide applicability as a cancer vaccine because it can target both T cell- and B cell-specific responses. It may thus represent a promising approach for the immunotherapy of colorectal cancer.

Immuno-stimulating Activities of Polysaccharide Fractions Isolated from Persimmon Leaves (감잎에서 분리한 다당의 면역자극 활성)

  • Shin, Young-A;Park, Hye-Ryung;Hong, Hee-Do;Shin, Kwang-Soon
    • The Korean Journal of Food And Nutrition
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    • v.25 no.4
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    • pp.941-950
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    • 2012
  • In order to develop new physiologically active polysaccharides from persimmon leaves, two different crude polysaccharides were prepared using hot water (PLW-0) and pectinase digestion (PLE-0) and their immuno-stimulating activities were estimated. PLW-0 and PLE-0 showed similar sugar compositions with 15 different sugars, including rarely observed sugars in general polysaccharides such as 2-O-methyl-fucose, 2-O-methyl-xylose, apiose, aceric acid, 3-deoxy-D-manno-2-octulosonic acid, and 3-deoxy-D-lyxo-2-heptulosaric acid, but the uronic acid content of PLE-0 was lower than that of PLW-0 caused by pectinase treatment. Both PLW-0 and PLE-0 showed potent anti-complementary activity in a dose-dependent manner which was similar to a known immuno-stimulating polysaccharide, PSK, from Coriolus versicolor. The activity of PLE-0 at a low concentration ($100{\mu}g/m{\ell}$) was higher than that of PLW-0. In an in vitro cytotoxicity analysis, PLW-0 and PLE-0 (up to $1,000{\mu}g/m{\ell}$) did not affect the growth of peritoneal macrophages and Colon 26-M3.1 carcinoma cells. In contrast, they enhanced lymphocyte proliferation activity. Peritoneal macrophages stimulated with PLW-0 and PLE-0 produced various cytokines, such as IL-6 and IL-12. However, PLE-0 was more effective on the cytokine production. Intravenous administration of PLW-0 and PLE-0 significantly augmented natural killer (NK) cell cytotoxicity against Yac-1 tumor cells 3 days after the treatment of polysaccharide fractions. But NK cells obtained from the PLE-treated group showed higher tumoricidal activity even at a low dose of $40{\mu}g$/mouse. In experimental lung metastasis of Colon 26-M3.1 carcinoma cells, prophylactic administration of PLW-0 and PLE-0 significantly inhibited lung metastasis in a dose-dependent manner and PLE-0 was more effective on the inhibition of cancer metasasis. The results lead us to conclude that the pectinase-treated process is indispensable to preparing polysaccharides with higher immune-stimulating activity from persimmon leaves.

Potentiating Dietary Green Tea Extracts Anti-Tumor Activity of Cisplatin in BALB/c Mice Bearing CT26 Colon Carcinoma (대장암(CT 26) 생쥐에서 녹차추출물 음용에 의한 시스플라틴 항암작용 증강효과)

  • Lee, Byoung-Rai;Park, Pyoung-Sim
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1100-1105
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    • 2012
  • Green tea intake is known to have preventive effects against cancer. In this study, we evaluated the tumor suppressive effects of dietary green tea extracts (GTE) as a modulator on cisplatin in an established colon cancer mouse model. The cisplatin-induced cytotoxicity was determined with cell viability of the mouse colon cancer cell line (CT26) in vitro. The influence of GTE on the anti-tumor activity of cisplatin was evaluated by measuring tumor size with digital calipers in mice bearing CT26 colon carcinomas. The CT26 cell viability decreased to 93% at a $20{\mu}g/mL$ concentration of cisplatin. However, cell viability decreased to 15% with a combination of $20{\mu}g/mL$ cisplatin and GTE ($75{\mu}g/mL$). There were no apparent changes in cisplatin-induced cytotoxicity with GTE and epigallocathechin gallate (EGCG) treatments. Tumor size decreased in dietary GTE combining intra-peritoneal cisplatin-injected tumors bearing mice compared with cisplatin or GTE alone administered to tumor-bearing mice. These experiments showed that dietary GTE has a potentiating effect on the cisplatin anti-tumor activity of an established mice colon cancer model. Therefore, the GTE may be a candidate for modulators in anticancer treatments with cisplatin.

Biocompatibility of experimental mixture of mineral trioxide aggregate and glass ionomer cement (Glass ionomer cement와 혼합한 mineral trioxide aggregate의 생체친화성)

  • Oh, Min-Jae;Jeong, Yu-Na;Bae, In-Ho;Yang, So-Young;Park, Bum-Jun;Koh, Jeong-Tae;Hwang, Yun-Chan;Hwang, In-Nam;Oh, Won-Mann
    • Restorative Dentistry and Endodontics
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    • v.35 no.5
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    • pp.359-367
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    • 2010
  • Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.

Biological Activities of Oenothera laciniata Extracts (Onagraceae, Myrtales) (애기달맞이꽃(Oenothera laciniata Hill) 추출물의 생리활성 탐색)

  • Lee, Jung-A;Kim, Ji-Young;Yoon, Weon-Jong;Oh, Dae-Ju;Jung, Yong-Hwan;Lee, Wook-Jae;Park, Soo-Yeong
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.810-815
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    • 2006
  • The biological activities of Oenothera laciniata extracts were measured, including antioxidant activity and cytotoxic effects. O. laciniata is an endemic species of Jeju Island, Korea with a seaside habitat. The concentration of total polyphenolic compounds from ethanol (EtOH), n-hexane, dichloromethane ($CH_2Cl_2$), ethylacetate (EtOAc), butanol (BuOH), and water fractions of O. laciniata was 63.96, 8.49, 28.11, 172.64, 114.56, and 34.91 mg/g, respectively. The EtOAc fraction contained the highest antioxidative activities ($IC_{50}$), measured as follows: 16.19 ${\mu}g/mL$ in DPPH radical scavenging capacity, 220.37 ${\mu}g/mL$ in xanthine oxidase inhibitory activity, 42.07${\mu}g/mL$ in superoxide radical scavenging capacity, and 421.33 ${\mu}g/mL$ in nitric oxide scavenging capacity. The cytotoxicity of O. laciniata extracts was examined through their effect on the growth of HL-60 cells. Incubation of HL-60 cells with the EtOAc fraction resulted in the greatest inhibition of cell growth; high DNA fragmentation and numerous sub-G1 hypodiploid cells were observed in HL-60 cell cultures treated with the EtOAc fraction. These results suggest that the EtOAc fraction of O. laciniata has potent apoptotic and antioxidative activities in vitro.

Antitumor and Immunomodulatory Effects of Chungsangbohahwan on Murine Melanoma-induced Lung Metastasis (청상보하환(淸上補下丸)이 흑색종(黑色腫)의 폐전이(肺轉移) 억제(抑制) 및 면역조절작용(免疫調節作用)에 미치는 영향(影響))

  • Ha, Jee-Yong;You, Byeong-Gil
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.159-175
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    • 1998
  • In order to investigate the antitumor effect by Chungsangbohahwan after B-16 cells were transplanted in C57BL/6 mice, and the immune responses in mice induced by methotrexate, the extract of Chungsangbohahwan was orally administered to the mice for 21 days. Experimental studies were performed for measurance of metastasis, cell cytotoxicity in vitro, natural killer cell activity, productivity of interleukin-2. The results were summarized as follows: 1. Inhibition of metastasis in Chungsangbohahwan-treated group was higher than control group with significance on 7th day and 14th day. 2. On the MTT assay, cell viability was significantly inhibited by $5{\mu}g/well$, $2.5{\mu}g/well$, $1.25{\mu}g/well$, and $0.625{\mu}g/well$ of Chungsangbohahwan concentration inhibited cell viability significantly(P<0.05). $IC_{50}$ for cell viability was $2.17{\mu}g/well$. 3. Natural killer cell activity in Chungsangbohahwan-treated group was significantly increased on 100:1, 50:1 E/T(effect cell/target cell) ratio(P<0.05). 4. Production of interleukin-2 in Chungsangbohahwan-treated group was significantly increased(P<0.05).

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