• Title/Summary/Keyword: MTT and SRB assay

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Increased Antimutagenic and Anticancer Activities of Chinese Cabbage Kimchi by Changing Kinds and Levels of Sub-Ingredient (부재료 첨가 배추김치의 항돌연변이 및 항암성 증진효과)

  • 박건영;조은주;이숙희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.4
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    • pp.625-632
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    • 1998
  • To enhance the antimutagenic and anticancer activities of chinese cabbage kimchi, 13 kinds of kimchi, which were different kinds and levels of sub-ingredient added kimchi, were prepared and fermented at 15℃ for 1 day and then at 5℃ up to pH 4.3. The antimutagenic effects of the methanol extracts of the kimchi were studied by using Ames mutagenicity test in Salmonella typhimurium TA100 and SOS chromotest in E. coli PQ37. Among the kimchi samples, high ratio of red pepper powder(7%) and garlic(2.8% or 5.2%) added kimchi, 1% chinese pepper powder added kimchi and organic cultivated chinese cabbage kimchi significantly reduced(p<0.05) the mutagenicity induced by aflatoxin B1(AFB1) in Ames test and SOS response against N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in SOS chromotest, and also the kimchi inhibited more effectively(p<0.05) the survival and growth of AGS human gastric adenocarcinoma cells than the standarized kimchi on the SRB assay, MTT assay and growth inhibition test. These results suggest that the antimutagenic and anticancer activites of kimchi can be increased by the sub-ingredients such as organic cultivated chinese cabbage, red pepper powder, garlic and chinese pepper powder.

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Neurotoxicity of Sodium Salicylate on Spinal Sensory Neurons in Culture (배양 척수감각신경세포에 대한 살리실산 나트륨의 신경독성에 관한 연구)

  • Lee, Kang-Chang;Choi, Yu-Sun;Park, Seung-Taeck
    • The Korean Journal of Pain
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    • v.14 no.2
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    • pp.136-141
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    • 2001
  • Background: Sodium salicylate (SS) is a nonsteroidal anti-inflammatory drug (NSAID) for the treatment of neuralgia or pain from rheumatoid arthritis. When abused or used in excess, SS can induce cytotoxicity. The present study examined whether SS has a neurotoxic effect. Methods: Cell viability was examined by MTT [3-(4,5-dimethylthiazol-2,5-dipheny ltetrazolium bromide] assay and Sulforhodamine (SRB) assay after cultivating dorsal root ganglion (DRG) neurons derived from neonatal mouse. These cells were treated with various concentrations of SS for 24 hours. In addition, the amount of protein synthesis against SS was measured in these cultures. Results: Cell viability (20, $40{\mu}g/ml$ SS) significantly decreased in a dose-dependent manner. Additionally, SS inhibited protein synthesis after the exposure of cultured mouse DRG neurons to $30{\mu}g/ml$ of SS for 24 hours. Conclusions: The present study suggests that SS is toxic in cultured DRG neurons derived from neonatal mouse by decreasing cell viability and the amount of protein synthesis.

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Influence of Kamijihwang-hwan on the Hypoxic Damage of Cultured Cerebral Neurons from mouse and SK-N-MC cells (가미지황환이 저산소성 신경세포 손상에 미치는 영향)

  • Kyung Baek Yeun;Ju Sung Min;Kim Kun Jun;Kim Dae Keun;Kang Jeong Ho;Lee Young Chan;Lee Jun;Kim Young Mok;Jeon Byung Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.1082-1091
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    • 2003
  • To elucidate the neuroprotective effect of Kamijihwang-hwan(KSH) on nerve cells damaged by hypoxia, the cytotoxic effects of exposure to hypoxia were determined by XTT, NR, MTT and SRB asssay. The activity of catalase and SOD was measured by spectrophometry, and TNF-α and PKC activity was measured after exposure to hypoxia and treatment of Kamijihwang-hwan(KSH) water extract(KJHWE). Also the neuroprotective effect of KJHWE was researched for the elucidation of neuroprotective mechanism. The results were as follows ; Hypoxia decreased cell viability measured by XTT, NR assay when cultured cerebral neurons were exposed to 95% N2/5% CO₂ for 2~26 minutes in these cultures and KJHWE inhibited the decrease of cell viability. H₂O₂ treatment decreased cell viability measured by MTT, and SRB assay when cultured cerebral neurons were exposed to 1-80 uM for 6 hours, but KJHWE inhibited the decrease of cell viability. Hypoxia decreased catalase and SOD activity, and also TNF-α and PKC activity in these cultured cerebral neurons, but KJHWE inhibited the decrease of the catalase and SOD activity in these cultures. Hypoxia triggered the apoptosis via caspase activation and internucleosomal DNA fragmentation. Also hypoxia stimulate the release of cytochrome c form mitochondria. KJHWE inhibited the apoptosis via caspase activation induced by hypoxia. From these results, it can be suggested that brain ischemia model induced hypoxia showed neurotoxity on cultured mouse cerebral neurons, and the KJHWE has the neuroprotective effect in blocking the neurotoxity induced by hypoxia in cultured mouse cerebral neurons.

Influence of Yeoldahanso-tang on the Hypoxic Damage of Cultured Cerebral Neurons from mouse and SK-N-MC cells (열다한소탕(熱多寒少湯)이 저산소성(低酸素性) 대뇌신경세포(大腦神經細胞) 손상에 미치는 영향(影響))

  • Kim, Hyoung-Soon;Bae, Young-Chun;Lee, Sang-Min;Kim, Kyung-Yo;Won, Kyoung-Sook;Sihm, Gyue-Hearn;Park, Su-Jeong
    • Journal of Sasang Constitutional Medicine
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    • v.15 no.1
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    • pp.72-89
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    • 2003
  • To elucidate the neuroprotective effect of Yeoldahanso-tang(YHT) on nerve cells damaged by hypoxia, the cytotoxic effects of exposure to hypoxia were determined by XTT(SODIUM3,3'-{I-[(PHENYLAMINO) CARBONYL]-3,4-TETRAZOLIUM}- BIS (4-METHOXY-6-NITRO) BENZENE SULFONIC ACID HYDRATE), NR(Neutral red), MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and SRB(Sulforhodamin B) asssay. The activity of catalase and SOD(Superoxide dismutase) was measured by spectrophometry, and $TNF-{\alpha}$(Tumor cell necrosis $fector-{\alpha}$) and PKC(Protein kinase C) activity was measured after exposure to hypoxia and treatment of YHTWE. Also the neuroprotective effect of YHTWE was researched for the elucidatioion of neuroprotective mechanism. The results were as follows; 1. Hypoxia decreased cell viability measured by XTT, NR assay when cultured cerebral neurons were exposed to 95% N2/5% CO2 for $2{\sim}26$ minutes in these cultures and YHTWE inhibited the decrease of cell viability. 2. H2O2 treatment decreased cell viability measured by MTT, and SRB assay when cultured cerebral neurons were exposed to 1-80 ${\mu}M$ for 6 hours, but YHTWE inhibited the decrease of cell viability. 3. Hypoxia decreased catalase and SOD activity, and also $TNF-{\alpha}$ and PKC activity in these cultured cerebral neurons, but YHTWE inhibited the decrease of the catalase and SOD activity in these cultures. 4. Hypoxia triggered the apoptosis via caspase activation and internucleosomal DNA fragmentation. Also hypoxia stimulate the release of cytochrome c forom mitochondria. YHTWE inhibited the apoptosis via caspase activation induced by hypoxia. From these results, it can be suggested that brain ischemia model induced hypoxia showed neurotoxicity on cultured mouse cerebral neurons, and the YHTWE has the neuroprotective effect in blocking the neurotoxicity induced by hypoxia in cultured mouse cerebral neurons.

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Effects of Gamibojungikki-tang on Total Protein Synthesis of Cultured Spinal Sensory Neurons Damaged by GLUCOSE OXIDASE (가미보중익기탕이 GLUCOSE OXIDASE에 의해 손상된 배양 척수감각신경세포의 총단백질 합성량에 미치는 영향)

  • Ho Lee Chang;Beam Kwon Kang;Ho Jang Seung;Sun Song Yong;Gon Ryu Do
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.1
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    • pp.141-145
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    • 2002
  • In order to clarify the neuroprotective effect of Gamibojungikki-tang (GBJIKT) water extract on cultured mouse spinal sensory neuron damaged by glucose Oxidase (GO), MTT [3-(4,5-dimethylthiazole-2-yl) -2,5-diphenyltetrazolium bromide] assay and SRB (Sulforhodamine B) assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of GBJIKT water extract for 3 hours prior to exposure of GO. Cell viability of cultured mouse spinal sensory neurons exposed to various concentrations of GO for 8 hours was decreased in a dose-dependent manner. MTT50 values were 45 mU/ml GO. Cultured mouse spinal sensory neurons in the medium containing various concentration of GO for 8 hours showed decreasing of total protein synthesis. GO was toxic on cultured spinal sensory neurons. Pretreatment at GBJIKT water extract for 3 hours following GO prevented the GO-induced neurotoxicity such as decreasing of total protein synthesis. These results suggest that GO shows toxic effect on cultured spinal sensory neurons and GBJIKT water extract is highly effective in proecting the neurotoxicity induced by GO.

Orthosiphon pallidus, a Potential Treatment for Patients with Breast Cancer

  • Singh, Mukesh K.;Dhongade, Hemant;Tripathi, Dulal Krishna
    • Journal of Pharmacopuncture
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    • v.20 no.4
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    • pp.265-273
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    • 2017
  • Objective: Orthosiphon pallidus (O. pallidus), which belongs to the Lamiaceae family, is a popular garden plant that is widely used for the treatment of various diseases, such as urinary lithiasis, fever, hepatitis, cancer and jaundice. The objective of the present work was to investigate the antioxidant free-radical scavenging and the anticancer activities of O. pallidus against human breast-cancer cell lines. Methods: The antioxidant activity of Orthosiphon pallidus aqueous extract (OPAE) was investigated using different models, such as the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the 2, 2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) models, as were the $Fe^+$ chelation, the hydroxyl radical and superoxide radical scavenging, and total reducing power activities. The anticancer activities of the extract were determined by using the 3-(4, 5- dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) and the sulforhodamine (SRB) assays on the MCF-7 and the MDA-MB-231 cancer cell lines. Results: The aqueous Orthosiphon pallidus extract showed potent activity in in-vitro models. It significantly inhibited the scavenging of hydroxyl and superoxide radicals, but induced a remarkable $Fe^+$ chelation activity. For both cell lines, the percent cytotoxicity was found to increase steadily with increasing OPAE concentration up to $240{\mu}g/mL$. Conclusion: These results suggest that Orthosiphon pallidus has excellent antioxidant, antimicrobial, and anticancer activities against human breast-cancer cell lines.

Streptomyces endus YP-1이 생산하는 항암활성 물질의 분리 및 정제

  • Choi, Sung-Won;Kim, Byoung-Chan;Choi, Sun-Jin;Kim, Dong-Seob;Yeo, Ick-Hyun;Moon, Soon-Ok;Oh, Doo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.25 no.5
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    • pp.483-489
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    • 1997
  • Sulforhodamine B (SRB) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, RNA dot blot and Northern hybridization analysis were performed to screen microorganisms for the production of anticancer agent. Among microorganisms tested, strain YP-1 was selected for its cytotoxicity and ability to reduce the level of c-myc RNA. Strain YP-1 was identified as Streptomyces endus. The anticancer material produced by Streptomyces endus YP-1 was sequentially purified by solvent extraction, silica gel column chromatograpby, preparative TLC and preparative HPLC. The cancer material identified as azalomycin B by the instrumental analyses such as $^{1}$H-NMR, $^{13}$C-NMR, Mass, IR and UV absorption. It was colorless amorphous powder and its molecular weight was 1025.278. Azalomycin B, produced by Streptomyces endus YP-1, showed anticancer activity against several human cancer cell lines and reduction of c-Myc protein level in Colo320 DM cells which was determined by Western blot analysis.

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Studies on Immunomodulatory and Antioxidant Activities of Astragali membranacei Radix according to the Cultivated Years (황기(黃芪)의 재배 년수에 따른 면역 및 항산화 활성 연구)

  • Jung, Chul
    • Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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    • v.1 no.1
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    • pp.53-90
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    • 2005
  • Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

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EFFECT OF OXYGEN TENSION ON THE ACTIVITY AND FUNCTION OF THE CELLS DERIVED FROM HUMAN PERIODONTAL LIGAMENT (산소 장력이 치주인대에서 유래한 세포의 활성과 기능에 미치는 영향)

  • Sa, Muyng-Hee;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.27 no.3 s.62
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    • pp.503-513
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    • 1997
  • This study was undertaken to investigate the effect of oxygen tension on the activity and function of the cells derived from human periodontal ligament by measuring cell activity, total protein synthesis, collagen synthesis, $IL-1{\beta},\;IL-6,\;TNF-{\alpha}$ Human periodontal ligament fibroblasts were collected from premolars extracted for orthodontic treatment and incubated in the environment of $37^{\circ}C,\;5\%\;CO_2,\100\%$ humidity. After the fifth to sixth passage they were used for the experiment. Gaspack system to which $0.2{\mu}m$ Millipore filter was attached was connected to mixed-gas tanks. The mixed gases were composed of $10\%\;O_2,\;5\%\;CO_2,\;85\%\;N_2$ in hyoxic group or $90\%\;O_2,\;5\%\;CO_2,\;5\%\;N_2$ in hyperoxic group and $5\%\;CO_2,\;95\%$ air for control. After incubation in $37^{\circ}C$ for 2, 4, 6 days, cell activity was determined by tetrazolium(MTT) assay and total protein synthesis was assayed using sulforhodamine B(SRB). And measurement of 4-hydroxyproline was performed to assess collagen synthesis md $IL-1{\beta},\;IL-6,\;and\;TNF-{\alpha}$ were measured by enzymeimmunoassay. The results were as follows. 1. The cell activity and total protein synthesis in hypoxic group were a little higher than or almost the same with those in control group. 2. In hyperoxic group, the cell activity was lower than that in control group and total protein synthesis was decreased. 3. Collagen synthesis was significantly decreased initially in both hypoxic and hyperoxic group and increased nearly to the level of control group as the duration of cell incubation was longer 4. As a result of enzymeimmunoassay, the amount of cytokines was $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ in order. 5. $IL-6,\;TNF-{\alpha}\;and\;IL-1{\beta}$ were increased more rapidly in both hypoxic and hyperoxic group than in control group as the duration of cell incubation was longer. 6. There were more $IL-6\;and\;TNF-{\alpha}$ in hyperoxic group than in control group after 6 days, and there were more $IL-6\;and\;TNF-{\alpha}$ after 6 days than after 2 or 4 days in hyperoxic group. These results suggested that oxygen tension might modulate the production of extracellular matrix and cytokines in the cells derived from human periodontal ligament.

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Comparison of Contact Lenses and Storage Solution for Cytotoxic Potential using a Cell Growth Inhibition Assay (소프트 콘택트렌즈 용출액과 보존 용액의 세포 독성 비교)

  • Kim, Jai-Min;Seong, Jeong-Sub;Yoo, Keun-Chang;Yoon, Young;Ra, Myung-Suk;Lee, Jong-Bin
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.2
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    • pp.81-86
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    • 1999
  • Human corneal epithelial cells and mouse fibroblast L929 cells were grown to 60-70% confluency in 96 well plates. 24 hours after the well plates are inoculated, the medium on the test plates is aspirated and replaced with an extract supplemented medium prepared from the materials to be tested. The contact lenses manufactured by 7 companies were collected from optical shops and used for this study. The exactracts having been prepared by autoclaving 8 lenses in 2.5ml saline. The cell monolayer is then cultured for a further 48 hour period. MTT and SRB assys were performed for cytotoxic effect on cultured cells An inhibition of 30% is considered clear indication of cytotoxic potential in the test material. All the materials were not cytotoxic, but 3 storage solutions of them inhibited growing L929 cells.

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