• 제목/요약/키워드: Rat2 cell line

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Genotoxicity Study of Sophoricoside, a Constituent of Sophora japonica, in Bacterial and Mammalian Cell System

  • Kim, Youn-Jung;Park, Hyo-Joung;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.99-105
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    • 2001
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to has an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial reversion test, chromosomal aberration assay and single cell gel electrophoresis (Comet) assay. As results, in the range of 1,250~40 $\mu\textrm{g}$/plate sophoricoside concentrations was not shown significant mutagenic effects in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 strains in Ames test. The 80% cell growth inhibition concentration (IC/SUB 80/) of sophoricoside was determined as above 5,000 $\mu\textrm{g}$/$m\ell$ in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line for the chromosomal aberration and comet assay, respectively. Sophoricoside was not induced chromosomal aberration in CHL fibroblast cell at concentrations of 700, 350 and 175 $\mu\textrm{g}$/$m\ell$ or 600, 300 and 150 $\mu\textrm{g}$/$m\ell$ in the absence or presence of S-9 metabolic activation system, respectively. Also, in the comet assay, the induction of DNA damage was not observed in L5178Y mouse lymphoma cell line both in the absence or presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside were observed in bacterial and mammalian cell systems used in these experiments.

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A New Bioluminescent Rat Prostate Cancer Cell Line: Rapid and Accurate Monitoring of Tumor Growth (효과적인 항암효능측정을 위한 발광 전립선 세포의 개발 및 평가)

  • Lee, Mi-Sook;Jung, Jae-In;Kwon, Seung-Hae;Shim, In-Sop;Hahm, Dae-Hyun;Han, Jeong-Jun;Han, Dae-Seok;Yoonpark, Jung-Han;Her, Song
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1738-1741
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    • 2010
  • Caliper measurements of tumor volume have been widely used in the assessment of tumors in animal models. However, experiments based on caliper data have resulted in unreliable estimates of tumor growth, due to necrotic areas of tumor mass. To overcome this systematic bias, we engineered a new luciferase-expressing rat prostate cancer cell line (MLL-Luc) that produces bioluminescence from viable cancer cells. MLL-Luc cells showed a strong correlation between bioluminescence intensity and cell number ($R^2$=0.99) and also accurately quantified tumor growth, with reduced bioluminescence signals caused by necrotic cells in a subcutaneous MLL-Luc xenograft model. The accurate quantification of tumor growth with bioluminescence imaging (BLI) was confirmed by a better antitumor effect of combination chemotherapy, compared to that based on caliper measurements with a correlation between the bioluminescence signal and tumor volume ($R^2$=0.84). These data suggest that bioluminescent MLL xenografts are a powerful and quantitative tool for monitoring tumor growth and are useful in evaluating the efficacy of anticancer drugs, with less systematic bias.

Flow Visualization of Blood Cell and Detection of Cell Depleted Layer Using a Confocal Laser Scanning Microscope (공초점 레이저 주사 현미경을 이용한 혈구 유동가시화 및 세포공핍층 측정에 관한 연구)

  • Lim, Soo-Hee;Kim, Wi-Han;Lee, Ho;Lee, Choon-Young;Park, Cheol-Woo
    • Journal of the Korean Society of Visualization
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    • v.8 no.1
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    • pp.46-52
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    • 2010
  • In the present study, we employed the confocal laser scanning microscopy (CLSM) system to visualize the blood flow field with $1{\times}1{\mu}m^2$ spatial resolution. Based on the confocal microscopic image of red blood cells (RBCs), we performed the velocity vector field measurement and evaluated characteristics of cell migration from the cell depleted layer thickness calculation. The rat and mouse's blood were supplied into a micro glass tubes in vitro. The line scanning rate of confocal microscopy was 15 kHz for a $500{\times}500$ pixels image. As a result, the red blood cell itself can be used as a tracer directly without any kind of invasive tracer particle to get the velocity vector field of blood flow by performing particle image velocimetry (PIV) technique.

Experimental Studies on Antitumor Activity of Herb Drugs (I)-Effectiveness on Rat Natural Killer Cell Activity- (수종(數種)의 생약(生藥)에 대(對)한 항암효과(抗癌效果)의 실험적(實驗的) 연구(硏究)(I) -백서(白鼠)의 자연살해세포활성(自然殺害細胞活性)에 미치는 영향(影響)-)

  • Kang, Yun-Ho;Kim, Byung-Woon;Ha, Youn-Mun;Park, Jai-Kyung;Nam, Sang-Yun;Choi, Kyu-Chul;Choi, Yong-Mook
    • Korean Journal of Pharmacognosy
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    • v.18 no.2
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    • pp.118-126
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    • 1987
  • Natural Killer cells are considerd to play an important role in antitumor immune surveilance mechanism. In this study, 21 putative anticancer drugs selected from reference were assessed by evaluating the effect on rat Natural Killer cell activity (NKCA). All 21 herb drugs were extracted in boiling water, lyophilized, autoclaved, and then used for experiment. Culture supernatant of concanavalin-A (Con-A)-stimulated rat spleen cells as a source of lymphokine was also used as a control of comparison. Rat spleen cells were used as effector and NKCA was measured in 4hr $^{51}Cr-release$ assay against Yac-1 mouse lymphoma cell line. In order to determine the optimal conditions for NKCA augmentation, effector cells were treated with 3 different concentrations of each drug for 24, or 48 hrs before testing of NKCA, In optimal conditions determined from previous results, the effect of herb drugs on NKCA were assessed in 3 to 5experiments. NKCA was significantly enhanced by treatment with 4 herb drugs(Ponciri Fructus, Houttuyniae Herba, Aurantii Pericarpium, Nepetae Herba). Culture supernatant of Con-A-stimulated spleen cells also augmented the rat NKCA more significantly. The results show that 4 of the herb medicines supposed to display anticancer effect may have activity as a biological response modifier through augmentation of NKCA.

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A STUDY OF MORPHOGENESIS OF DIGITAL MALFORMATION ON RAT EMBRYO BY X-IRRADIATION (방사선조사시 태내백서의 지지기형성과정에 관한 실험적 연구)

  • Khim Jhai Dhuck
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.11 no.1
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    • pp.33-40
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    • 1981
  • The author studied on the effects of x-irradiation to the development of digital malformation in gestation rats. The time-matings occured between 6 p.m. and 8 a.m. and females with copulation. plugs at 8 a.m. were isolated and properly marked for evidence of copulation. The lower abdomen of mothers were exposed to x-irradiation on the 11½th day of gestation, the critical period developing digital malformation, respectively 100, 150, 200, 250, 300 and 350 rads. At 18½th day of post-conception total 50 pregnant females were dissected and the incidence of digital malformations were obtained. Rat embryos on the 12, 13, 14, 15, 16th day of gestation irradiated by 250 rads were examined for morphogenesis of digital malformation. Digital radiating lines were examined in water and histologically by H-E stain. Supra vital stain samples by Nile-blue sulfate in 37℃ normal saline were prepared for the observation of cell necrosis regions and morphogenesis of digits. The results obtained were as follows; 1. By x-irradiation on 11th day of gestation, digital malformations of Ectrodactylia, Syndactylia Polydactylia and Hematodactylia were developed. Ectrodactylia showed the effective relationship to the amount of irradiation, however Syndactylia ans Polydactylia did not. 2. By x-irradiation, cell necrosis of digital germ was appeared markedly, but in 48 hours after irradiation was depressed to the periphery of digital germ and in 72hours after irradiation was disappeared. Digital radiating line showed marked stage of malformation in 48hours after irradiation and continued to show the same amount of physiological cell necrosis as the compared control group in 72hours. after irradiaion. But in the Syndactylia, physiological cell necrosis was not able to be recognized. 3. Ectrodactylia induced by x-irradiation was considered as the direct resoult of cell necrosis of digital origin, however, Polydactylia and Syndactylia were considered as the resoult of some effect in repair process of x-irradiation damages.

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Preventive effects of nano-graphene oxide against Parkinson's disease via reactive oxygen species scavenging and anti-inflammation

  • Hee-Yeong Kim;Hyung Ho Yoon;Hanyu Seong;Dong Kwang Seo;Soon Won Choi;Jaechul Ryu;Kyung-Sun Kang;Sang Ryong Jeon
    • BMB Reports
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    • v.56 no.3
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    • pp.202-207
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    • 2023
  • We investigated the neuroprotective effects of deca nano-graphene oxide (daNGO) against reactive oxygen species (ROS) and inflammation in the human neuroblastoma cell line SH-SY5Y and in the 6-hydroxydopamine (6-OHDA) induced Parkinsonian rat model. An MTT assay was performed to measure cell viability in vitro in the presence of 6-OHDA and/or daNGO. The intracellular ROS level was quantified using 2',7'-dichlorofluorescein diacetate. daNGO showed neuroprotective effects against 6-OHDA-induced toxicity and also displayed ROS scavenging properties. We then tested the protective effects of daNGO against 6-OHDA induced toxicity in a rat model. Stepping tests showed that the akinesia symptoms were improved in the daNGO group compared to the control group. Moreover, in an apomorphine-induced rotation test, the number of net contralateral rotations was decreased in the daNGO group compared to the control group. By immunofluorescent staining, the animals in the daNGO group had more tyrosine hydroxylase-positive cells than the controls. By anti-Iba1 staining, 6-OHDA induced microglial activation showed a significantly decrease in the daNGO group, indicating that the neuroprotective effects of graphene resulted from anti-inflammation. In conclusion, nano-graphene oxide has neuroprotective effects against the neurotoxin induced by 6-OHDA on dopaminergic neurons.

Effects of Unripe Black Raspberry Extracts on Prostate Cancer Cell Line and Rat Model of Benign Prostatic Hyperplasia (복분자 미숙과 추출물이 전립선암 세포주와 전립선비대 백서모델에 미치는 영향)

  • Lee, Su Jung;Choi, Hye Ran;Lee, Jung-Hyun;Kwon, Ji Wung;Lee, Hee Kwon;Jeong, Jong Tae;Lee, Tae-Bum
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.507-515
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    • 2014
  • Benign prostatic hyperplasia (BPH), which is commonly found in aging men, is characterized by hyperplasia of prostatic stromal and epithelial cells beginning in the periurethral zone of the prostate. The prevalence of BPH increases in an age-dependent manner. Here, we investigated the protective effects of unripe Rubus occidentalis extracts (UROE) on BPH development using a prostate cancer cell line and testosterone-induced BPH rat model. Experiments using an established hormone-dependent prostate cancer cell line (LNCaP) showed that UROE treatment significantly decreased expression of androgen-related genes, including androgen receptor (AR), prostate specific antigen (PSA), and 5-alpha reductase 2, but not 5-alpha reductase 1, which was also observed in flutamide-treated cells. Further, AR and PSA gene expression was reduced by UROE treatment under androgen-stimulated conditions using dihydrotestosterone (DHT). BPH animals displayed elevated prostate weights. However, UROE as well as finasteride treatment significantly reduced prostate weights and DHT levels compared to testosterone-induced BPH animals. Histopathological analysis also showed that UROE treatment suppressed testosterone-induced prostatic hyperplasia. Taken together, the results suggest that UROE may effectively inhibit the development of BPH and thus may be a useful agent in BPH treatment.

Differential Gene Expression in GPR40-Overexpressing Pancreatic ${\beta}$-cells Treated with Linoleic Acid

  • Kim, In-Su;Yang, So-Young;Han, Joo-Hui;Jung, Sang-Hyuk;Park, Hyun-Soo;Myung, Chang-Seon
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.2
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    • pp.141-149
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    • 2015
  • "G protein-coupled receptor 40" (GPR40), a receptor for long-chain fatty acids, mediates the stimulation of glucose-induced insulin secretion. We examined the profiles of differential gene expression in GPR40-activated cells treated with linoleic acid, and finally predicted the integral pathways of the cellular mechanism of GPR40-mediated insulinotropic effects. After constructing a GPR40-overexpressing stable cell line (RIN-40) from the rat pancreatic ${\beta}$-cell line RIN-5f, we determined the gene expression profiles of RIN-5f and RIN-40. In total, 1004 genes, the expression of which was altered at least twofold, were selected in RIN-5f versus RIN-40. Moreover, the differential genetic profiles were investigated in RIN-40 cells treated with $30{\mu}M$ linoleic acid, which resulted in selection of 93 genes in RIN-40 versus RIN-40 treated with linoleic acid. Based on the Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG, http://www.genome.jp/kegg/), sets of genes induced differentially by treatment with linoleic acid in RIN-40 cells were found to be related to mitogen-activated protein (MAP) kinase- and neuroactive ligand-receptor interaction pathways. A gene ontology (GO) study revealed that more than 30% of the genes were associated with signal transduction and cell proliferation. Thus, this study elucidated a gene expression pattern relevant to the signal pathways that are regulated by GPR40 activation during the acute period. Together, these findings increase our mechanistic understanding of endogenous molecules associated with GPR40 function, and provide information useful for identification of a target for the management of type 2 diabetes mellitus.

Transport of choline and its relationship to transport of cationic drugs in immortalized rat brain capillary endothelial cell line

  • Park, Hong-Mi;Lee, Kyeong-Eun;Lee, Na-Young;Kang, Young-Sook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.106-106
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    • 2003
  • Choline serves critical roles in the CNS both as a precursor of neurotransmitter and as an essential component of membrane phospholipids. The long-term maintenance of brain choline concentration is dependent on choline transport across the blood-brain barrier (BBB), And, we examined to elucidate the characteristics of transport of choline across the BBB using conditionally immortalized rat brain capillary endothelial cell line (TR-BBB) in vitro. The [$^3$H]choline in TR - BBB was increased by time dependently, but independent on Na$\^$+/, and the transport process is saturable with Michaelis-Menten constrant, Km of about 26 ${\mu}$M. The uptake of [$^3$H]choline is susceptible for inhibition by various organic cationic compounds including hemicholinium-3, tetraethylammonium chloride (TEA) and $\ell$-carnitine. Also, we investigated the relationship of transport of choline and cationic drugs. The uptake of [$^3$H]choline is inhibited by antioxidant, a-phenyl-n-tert-butyl nitrone (PBN) with IC$\sub$50/ of 1.2 mM. and by Alzheimer's disease therapeutics, such as acetyl $\ell$-carnitine, tacrine and donepezil. Also, choline uptake presented competitive inhibition with PBN, donepezil and acetyl $\ell$-carnitine in Lineweaver-Burk plot. In conclusion, TR-BBB cells express a saturable transport system for uptake of choline, and several cationic drugs may be transported into the brain by BBB choline transporter.

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Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes

  • Kim, Min Jae;Jung, Bong-Kwang;Cho, Jaeeun;Song, Hyemi;Pyo, Kyung-Ho;Lee, Ji Min;Kim, Min-Kyung;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.54 no.2
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    • pp.147-154
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    • 2016
  • Toxoplasma gondii infection induces alteration of the host cell cycle and cell proliferation. These changes are not only seen in directly invaded host cells but also in neighboring cells. We tried to identify whether this alteration can be mediated by exosomes secreted by T. gondii-infected host cells. L6 cells, a rat myoblast cell line, and RH strain of T. gondii were selected for this study. L6 cells were infected with or without T. gondii to isolate exosomes. The cellular growth patterns were identified by cell counting with trypan blue under confocal microscopy, and cell cycle changes were investigated by flow cytometry. L6 cells infected with T. gondii showed decreased proliferation compared to uninfected L6 cells and revealed a tendency to stay at S or G2/M cell phase. The treatment of exosomes isolated from T. gondii-infected cells showed attenuation of cell proliferation and slight enhancement of S phase in L6 cells. The cell cycle alteration was not as obvious as reduction of the cell proliferation by the exosome treatment. These changes were transient and disappeared at 48 hr after the exosome treatment. Microarray analysis and web-based tools indicated that various exosomal miRNAs were crucial for the regulation of target genes related to cell proliferation. Collectively, our study demonstrated that the exosomes originating from T. gondii could change the host cell proliferation and alter the host cell cycle.