• Title/Summary/Keyword: NR assay

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Effects of Sintongchukeo-tang on the Cultured Spinal Sensory Neurons Injured by Hydrogen Peroxide (신통축어탕(身痛逐瘀湯)이 Hydrogen Peroxide에 의해 손상(損傷)된 배양(培養) 척수감각신경세포(脊髓感覺神經細胞)에 미치는 영향(影響))

  • Lee, Kye-Seung;Na, Young-Hoon;Cha, Yong-Suk;Heo, Yun;Kim, Do-Hwan;Han, Sang-Hyok;Park, Byong-Min;Lee, In;Moon, Byung-Soon
    • The Journal of Internal Korean Medicine
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    • v.22 no.4
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    • pp.557-565
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    • 2001
  • Objectives : This study was carried out to examine toxic effect of Sintongchukeo-tang on cultured mouse spinal sensory neurons inhibited by neurotoxicity induced by hydrogen peroxide. Methods : MTT assay, NR assay, LDH and neurofilament assay were performed after spinal sensory neurons were preincubater with various concentrations of Sintongchukeo-tang water extract before treatment of cells with hydrogen peroxide. Results : Hydrogen peroxide induced ceil degeneration such as the decrease of cell viability was measured by MTT and NR assay in the cultured mouse spinal sensory neurons. Sintongchukeo-tang water extract was effective in the decrease of LDH activities of neurons produced by hydrogen peroxide. Sintongchukeo-tang water extract was effective in the increase of amount of neurofilaments damaged by hydrogen peroxide. Conclusions : From the above results, it is suggested that hydrogen peroxide induces the inhibition of cell viability in cultured mouse spinal sensory neurons and Sintongchukeo-tang water extract was effective in cultured neurons damaged by hydrogen peroxide.

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Oxidative Stress in the Cell and Antioxidant Activity of Kalopanax Pictus Extracts (음나무 추출물의 세포 내 산화 스트레스와 항산화 활성)

  • Kim, Sea-Hyun;Park, Youngki;Jang, Yong-Seok;Han, Jingyu;Chung, Hun-Gwan
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.126-134
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    • 2007
  • This study reviewed the application of an extract from Kalopanax pictus stem bark and root bark as natural antioxidants. To investigate the effect on cell toxic level against transformed mouse fibroblast L929 in formula added with various extracts from Kalopanax pictus stem bark and root bark, this experiment was carried out by in vitro cytotoxicity method. Using DCFA-DA method, oxidative stress in cell was measured with other antioxidant activity methods including DPPH assay and NBT assay. Active oxygen inhibition rate for root bark insoluble hot water extracts showed the highest with 57.9% for 15 min treatment. In DPPH and NBT test, antioxidant activities of methanol extract from stem bark and insoluble hot water extract from stem bark were 96% (at 0.1%) and 95% (at 0.5%), respectively.

Effects of Jowiseungcheongtang water extract on Cultured Primary Hippocampal Cell Culture Damaged by XO/HX (조위승청탕(調胃升淸湯) 전탕액(煎湯液)이 XO/HX에 의해 손상(損傷)된 배양(培養) 해마신경세포(海馬神經細胞)에 미치는 효과(效果))

  • Kim, Hyoung-Soon;Kim, Kyung-Yo;Go, Gi-Deok;Kim, Il-Hwan;Song, Seung-Yun
    • Journal of Sasang Constitutional Medicine
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    • v.14 no.1
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    • pp.67-78
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    • 2002
  • Jowiseungcheongtang(JST) has been in Sasang constitution medicine for many years as a therapeutic agent for cerebral disease. But the effect of Jowiseungcheongtang(JST) on neurotoxicity is not known. The purpose of this study was to determine the effects of Jowiseungcheongtang(JST) on the hippocampal cell injured by Xanthine Oxidase/Hypoxanthine. The results were as follows: 1. XO/HX decreased the survival rate of the cultured hippocampal cells on NR assay and MTT assay. 2. JST water extract have efficacy of decreasing a amount of lipid peroxidation increased by XO/HX in cultured hippocampal cells. 3. JST water extract have efficacy of increasing DNA synthesis decreased by XO/HX in cultured hippocampal cells. From the above results, It is concluded that JST has marked efficacy in preventing cultured hippocampal cells from the damages by XO/HX.

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Effects of Taeyeumjoweetang against Glucose Oxidase-induced Neurotoxicity in the Cultured Mouse Cerebral Cortical Neurons (태음조위탕(太陰調胃湯)이 Glucose Oxidase에 의해 손상된 대뇌피질(大腦皮質) 신경세포(神經細胞)에 미치는 영향(影響))

  • Kim, Jong-kwan;Ryu, Do-gon;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.267-281
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    • 1999
  • 1. Purpose : The purpose of this study was to determine the effects of Taeyeumjoweetang on the cerebral neurons injured by glucose oxidase(GO). 2. Methods : I observed cell viability in mouse cerebral neurons exposed to glucose oxidase by NR assay and MTT assay and determined lipid peroxidation in mouse cerebral neurons exposed to glucose oxidase. After administration of Taeyeumjoweetang water extracts, I observed significant changes of cell viability, lipid peroxidation in mouse cerebral neurons exposed to glucose oxidase. 3. Results : GO induced cell degeneration such as the decrease of cell viability was measured by MTT and NR assay in the cultured mouse cerebral cortical neurons. Taeyeumjoweetang was effective in the increase of total protein of neurons inhibited by GO. Taeyeumjoweetang was effective in the decrease of lipid peroxidation of neurons produced by GO.

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Cytotoxic Effects of Extracts from Hairy Roots of Rheum undulatum L. (대황의 모상근 배양조직 추출물의 세포독성)

  • Hwang, Sung-Jin;Kim, Jae-Hun;Ra, Myung-Suk;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.1
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    • pp.8-14
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    • 2001
  • The purpose of this research was to investigate the effects of extracts from cultured hairy roots of R. undulatum on human kidney epithelial cells. The cytotoxicity was measured by colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) with human kidney epithelial cell lines A498. MTT, NR and SRB quantities decreased propotionally in cultured A498 cells treated with the water or chloroform extracts of cultured hairy roots at increasing concentrations. These results suggest that extracts of cultured hairy roots are cytotoxic on human epithelial cells. The cytotoxicity of chloroform fraction was stronger than that of water fraction. The values of $MTT_{50},\;NR_{50}\;SRB_{50}$ of the extracts of chloroform fraction and those of water fraction were measured to be $289.3{\mu}g/ml,\;302.7{\mu}g/ml,\;433.8{\mu}g/ml\;and\;475.8{\mu}g/ml,\;428.3{\mu}g/ml,\;549.5{\mu}g/ml$, in A498 cell line.

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The Effects of Woohwangcheongsim-won on Hypoxic E18 Cortical Neuroblast (우황청심원(牛黃淸心元)이 저산소증 유발 배양신경세포에 미치는 영향)

  • Cho, Gyu-Seon;Shin, Gil-Cho;Moon, Il-Soo;Lee, Won-Chul;Chung, Sung-Hyun
    • The Journal of Internal Korean Medicine
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    • v.22 no.2
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    • pp.135-144
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    • 2001
  • Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won on neuronal death of hypoxic E18 cortical neuroblast. Methods : To evaluate the effect of Woohwangcheongsim-won on neuronal death caused by hypoxia, the survival rate of E18 cortical neuroblast was measured with MTT assay and the changes of several synaptic proteins and enzymes were investigated with the immunoblot assays. Results : The E18 cortical neuroblasts were added 50, 100, 500, 1,000, and $5,000{\mu}g/ml$ Woohwangcheongsim-won. They showed neurotoxicity, when the concentration of Woohwangcheongsim-won was above $1,000{\mu}g/ml$. The E18 cortical neuroblasts, which were added 50, 100, and $500{\mu}g/ml$ Woohwangcheongsim-won, were exposed 98% $N_2/5%\;CO_2$ for 3 hours to induce hypoxia, 3 days later, the survival rate of $50{\mu}g/ml$ Woohwangcheongsim-won was 141.5% when compared to the control group. On the immuneblot assays, the expressions of ${\alpha}$CaMKII, NR2A, NR28, PDE2, PSD-95, and eEF-$1{\alpha}$ were increased in normoxia, but those of NR2A, NR2B were decreased in hypoxia when compared to the control group. Conclusions : The data shows that the effects of Woohwangcheongsim-won on neuronal death of hypoxic E18 cortical neuroblast is a significant result.

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Cytotoxicity of paraquat and compensatory effects of 3-methylcholanthrene in rat lung (Paraquat의 세포독성과 흰쥐의 폐에서 3-Methylcholanthrene의 독성경감효과)

  • Rim, Yo-Sup;Kim, Doc-Soo;Han, Du-Seok;Hwang, In-Taek
    • The Korean Journal of Pesticide Science
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    • v.6 no.2
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    • pp.96-104
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    • 2002
  • This study was carried out to investigate cytotoxicity of paraquat on NIH 3T3 fibroblasts, toxicity of paraquat and compensatory effects of 3-methylcholanthrene (3-MC) on the rat lung. In order to conduct MIT [3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl -2H-tetrazolium-bromide] and NR (Neutral red) assay, the $5.0{\times}10^4cell/ml$ of NIH 3T3 fibroblast in each well of 24 multi-dish were cultured. After 24 hours, the cells were treated with solution of paraquat (1, 25, 50 and $100{\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MIT and NR assay were performed to evaluate the cytotoxicity of cell organelles. $MTT_{50}\;and\;NR_{50}$ of paraquat were $1668.97{\mu}M\;and\;1030.85{\mu}M$, respectively. These $IC_{50}$ of Paraquat were decided as a low cytotoxicity by Borenfreund and Puemer (1984). In order to observe the toxicity and compensatory effects of paraquat on the rat lung, Spraque Dawley male rats were used as experimental animals and were divided into paraquat only treated group and simultaneous application group of paraquat and 3-MC, at 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment. The animals were sacrificed by decapitation and their or the lungs were immediately removed, immersed in fixatives, and were processed with routine method for light microscopic study. Paraffin sections were stained with H&E and iron hematoxylin of Verhoeff. Under the light microscopy, erythrocytes were full in alveolar capillaries at 3 hrs and congested at 24 hrs after paraquat administration. The great alveolar cells (Type II cell) were increased and mitosis of great alveolar were observed in interalveolar septa. Many lymphocytes, macrophages and polymorphonuclear (PMN) cells were observed in connective tissue surrounding lung tissue and germinal center in lymph follicles of terminal bronchiole. Alveolar macrophages were increased in interalveolar septa and alveoli at 48 hrs. And observed many alveolar macrophages at 96 hrs. In iron hematoxylin stain of Verhoeff, Collagen fiber were increased in respiratory bronchiole, interalveolar septa and alveoli and breath of alveoli, and alveolar pore were broaden. But, in paraquat plus 3-MC treated group, morphological changes were mild in lung tissue. These results indicate that 3-MC has a compensatory effects against toxicity of paraquat by conjugation with oxygen.

Development of Antitoxic Agents from Korean Medicinal Plants (Part 3) Repaired Effects of Methanol Fraction of Perilla Frutescens on 3T3 Fibroblast Treated with Cadmium (한국산 생약으로부터 해독물질의 개발(제3보) 카드뮴으로 처리한 3T3섬유모세포에서 소엽 메탄올 분획의 수복효과)

  • 한두석;유현경;곽정숙;백승화
    • Toxicological Research
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    • v.11 no.2
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    • pp.229-234
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    • 1995
  • This study was carried out to evaluate the cytotoxicity of cadmium on 3T3 fibroblast and to develop the antidote on 3T3 fibroblast which was injuried by $IC_{50}$ of cadmium. The groups for repaired effects were divided into 7 groups such as medium alone treated group. Cadmium treated $IC_{50}$ groups and 5 experimental groups $(IC_{50}$ cadmium plus $10^{-4}$ concentration of each methanol fraction). After incubation for 48 hrs in the same conditions, MTT (tetrazolium MTT), NR (neutral red) and SRB (sulforhodamine B protein) assay were measured. Light microscopic observations were also investigated. The ethyl acetate fraction of Perilla frutescens showed significantly repaired effect against cadmium cytotoxiclty and this fraction inhibited critical cell regeneration in light microscopy.

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A Study on the Antimicrobial Activity and in vitro Cytotoxicity of UVB Sunscreen Chemicals in Cosmetic Products (UVB 자외선 차단제의 항균력 및 피부자극에 관한 연구)

  • 최종완;허윤석;손근욱
    • Proceedings of the SCSK Conference
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    • 1992.09a
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    • pp.46-68
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    • 1992
  • To investigate the effect on the antimicrobial activity against S.aureus ATCC 6538, E.coli KCTC 1039 and cell toxic level against transformed mouse fibroblast L929 in formula added with various concentrations of UVB blockers commonly used in cosmetic products, these experiments were carried out by preservative efficacy testing methods and in vitro cytotoxicity methods. The results obtained were as follow ; 1) Octyl Dimethyl PABA had a broad antibacterial spectrum against the Gram (+) and the Gram(-) bacteria at 5.84 % concentration, but not Octyl Methoxycinnamate. 2) Antibacterial activity was decreased in a combined UVB blocker system of squalane base. Especially, Octyl Dimethyl PABA was inactivated by Octyl Methoxycinnamate at 5.84% concentration to a large extents , but not 4-Methylbenzylidene Camphor. 3) Within in vitro cytotoxicity by use of mouse fibroblast L929 on UV-B blockers, NR assay was more excellent than MTT assay on quantitative

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Carbofuran Cytotoxicity on Rat Fibroblast Cells and Compensation Effects of Phenobarbital Sodium and 3-Methycholanthrene on Its Toxicity (살충제 Carbofuran이 쥐의 NIH3T3 섬유모세포에 끼치는 독성 및 Phenobarbital Sodium과 3-Methylcholanthrene에 의한 보상효과)

  • Han, Seong-Soo;Rim, Yo-Sup
    • Korean Journal of Environmental Agriculture
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    • v.16 no.2
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    • pp.149-155
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    • 1997
  • This study was carried out to investigate the effects of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on carbofuran cytotoxicity and to develop antitoxic agents based on the effectivness. Experimental groups for carbofuran cytotoxicity were divided into five groups ; medium alone and four treatments of carbofuran (1, 25, 50 and $100{\mu}M)$, and those for compensation effects were divided into six groups ; medium alone, $IC_{50}$ carbofuran and four combinations of carbofuran and PB or 3-MC($IC_{50}$ carbofuran plus 1, 25, 50, $100{\mu}M$ of PB and 3-MC, respectively). After incubation for 48 hrs under the same conditions, MTT(Tetrazolium MTT), NR(Neutral red) and SRB(Sulforhodamine B protein) assay were performed. Fifty percentage inhibition of MTT, NR, and SRB against carbofuran in rat fibroblast cell were 60.7, 82.5 and $87.0{\mu}M$, respectively. At the combination treatments of $IC_{50}$ of carbofuran and $100{\mu}M$ of PB, the significant compensation effects were observed from the results of MTT and NR but not from that of SRB absorbance. And at the combination treatments of $IC_{50}$ of carbofuran and 3-MC, the relatively significant compensation effects were found at $50{\mu}M$ 3-MC from the results of MTT and at $100{\mu}M$ 3-MC from that of NR and SRB absorbances, respectively. From the results of light microscopy, combination treatments of $carbofuran(IC_{50})$ and PB or 3-MC showed good regeneration in carbofuran toxicity of rat fibroblast cells. These results suggest that PB or 3-MC can compensate the cytotoxity of carbofuran insecticide in rat NIH3T3 fibroblast cells.

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