• Title/Summary/Keyword: mRNA expression

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The Effect of Theophylline on Eotaxin mRNA Expression in Pulmonary Epithelial Cell Line A549 (인체 기도상피세포주에서 Theophylline에 의한 Eotaxin mRNA 발현억제)

  • Han, Min-Soo;Yoo, Jee-Hong;Kang, Hong-Mo
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.898-908
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    • 2000
  • Background : Eotaxin a CC chemokine specific for eosinophils, is implicated in the pathogenesis of asthma by recruiting eosinophils into the airways. Theophylline has been used for the treatment of asthma and recently was proposed to have an anti-inflammatory action. The aim of this study is to examine whether theophylline may inhibit the eosinophilic airway inflammation by reducing the expression of eotaxin. Methods : The expression of eotaxin mRNA was assessed by Northern analysis in A549 cells 4 h after stimulation with TNF-$\alpha$ or IL-1$\beta$. And then, theophylline was added to A549 cells stimulated with 0.1 ng/mL IL-1$\beta$. Results : Eotaxin mRNA expression rates induced by 0.1, 1, 10 ng/mL TNF-$\alpha$ as compared with $\beta$-actin, were 7%, 22%. 28%, respectively. Eotaxin mRNA expression rates induced by 0.01, 0.1, 1, 10 ng/mL IL-1$\beta$, as compared with $\beta$-actin, were 10%, 42%, 63%, 72%, respectively. Eotaxin mRNA expression rates after the addition of 0, 0.001, 0.01, 0.1 ${\mu}M$ dexamethasone induced by 10 ng/mL TNF-$\alpha$ as compared with $\beta$-actin, were 27%, 18%, 8%, respectively. Eotaxin mRNA expression rate after the addition of 0, 0.001, 0.01, 0.1 ${\mu}M$ dexamethasone induced by 0.1 ng/mL IL-1$\beta$ as compared with $\beta$-actin, were 43%, 47%, 12%, 8%, respectively. Eotaxin mRNA expression rates after the addition of 0, 0.001, 0.01, 0.1, 1, 10 mM theophylline induced by 0.1 ng/mL IL-1$\beta$, as compared with $\beta$-actin, were 48%, 40%, 33%, 22%, 16%, 14%, respectively. Conclusion : These results suggest that theophylline may reduce eosinophil infiltration of the airway at least in part by reducing the expression of eotaxin under the conditions of these experiments.

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Effect of Pedunculagin on IL-1$\beta$ mRNA Expression in Langerhans cells (랑게르한스세포에서 IL-1$\beta$ mRNA 발현에 대한 Pedunculagin의 효과)

  • 주성수;권희승;강희철;이도익
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.472-476
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    • 2002
  • Contact hypersensitivity (CHS) serves as a good model of cell-mediated reaction. Epidermal langerhans cell (LC) are thought to playa crucial role in the regulation of immune reaction of the skin, which elicit the CHS response by presenting Antigen to trafficking Ag-specific T cells within the skin. However, contact hypersensitivity is regarded as a negative side of immunities, caused by increased damaging immune response. Therefore, the study of effector molecule causing immune suppression is thought to be meaningful in the skin immune response. For this aim, this study investigated the influence of pedunculagin on cytokine, IL-$\beta$ expression from langerhans cell (LC). In vitro and in vivo, pedunculagin up-regulated the expression of IL-1$\beta$ mRNA. After PMA stimulation in vitro and DNFB sensitization in vivo, the expression of IL-1$\beta$ mRNA was down-regulated. This results suggested that pedunculagin could be immuno-modulator in skin immune system by modulating IL-1$\beta$ expression.

Hepatic Vascular Stress Gene Expression in the Liver Response to Trauma

  • Lee, Sun-Mee
    • Biomolecules & Therapeutics
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    • v.12 no.2
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    • pp.62-67
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    • 2004
  • Trauma remains one of the important sources leading to systemic inflammatory response anti sub-sequent multiple organ failure. Although hepatic microvascular dysfunction occurs during trauma, the mechanism responsible remains unclear. The aim of this study was to investigate the effect of trauma on hepatic vascular stress gene expression. Femur fracture (EFx) was induced by torsion to the femur at midshaft. Liver samples were taken for RT-PCR analysis of mRNA for gtenes of interest: endothelin-1 (ET-1), its receptors $ET_A$ and $ET_B$, nitric oxide synthases (iNOS and eNOS), cyclooxygenase-2 (COX-2), heme oxygenase-1 (HO-1), and tumor necrosis tactor-${\alpha}$ (TNF-${\alpha}$). The expression of ET-1 mRNA was significantly increased by FFx. Expression of mRNA in FFx group showed no change in $ET_A$, $ET_B$, iNOS and HO-1 and showed a slight increase of 2.2-fold and 2.7-fold for eNOS tll1d COX-2, respectively. The level of TNF-${\alpha}$ mRNA significantly increased in FFx group. In conclusion, mild trauma alone causes little change in expression of vasoactive mediators.

Construction of an RNase P Ribozyme Library System for Functional Genomics Applications

  • Hong, Sun-Woo;Choi, Hyo-Jei;Lee, Young-Hoon;Lee, Dong-Ki
    • Genomics & Informatics
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    • v.5 no.1
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    • pp.6-9
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    • 2007
  • An RNase P ribozyme library has been developed as a tool for functional genomics studies. Each clone of this library contains a random 18-mer and the sequence of M1 RNA, the catalytic subunit of RNase P. Repression of target gene expression is thus achieved by the complementary binding of mRNA to the random guide sequence and the successive target cleavage via M1 RNA. Cellular expression of the ribozyme expression was confirmed, and EGFP mRNA was used as a model to demonstrate that the RNase P ribozyme expression system can inhibit the target gene expression. The constructed RNase P ribozyme library has a complexity of $1.4\times10^7$. This novel library system should become a useful in functional genomics, to identify novel gene functions in mammalian cells.

Reduced Cytotoxicity by Repetitive mRNA Transfection in Differentiated Neurons

  • Seung Hwan Ko;Jin Sun Kang;Sang-Mi Kim;Eun-Hye Lee;Chang-Hwan Park
    • International Journal of Stem Cells
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    • v.16 no.1
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    • pp.117-122
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    • 2023
  • Background and Objectives: mRNA-based protein expression technology has been used to express functional proteins. We have previously generated dopamine neurons from rat-embryo derived neural precursor cells (NPCs) through repeated transfection of synthetic transcription factor mRNA encoding dopamine-inducible genes. However, NPCs began to die approximately 10 d post-transfection. In this study, we examined a long-term transfection protocol that did not affect cell viability. Methods and Results: Experiments were performed in eight groups sorted according to the start date of mRNA transfection. mRNA was transfected into NPCs daily for 21 d and live cell images of each group were recorded. NPCs which were differentiated for more than five days showed sustained gene expression and appreciable viability despite daily mRNA transfection for 21 d. Conclusions: Repeated mRNA transfection requires cells with a sufficient differentiation period.

Effects of Aluminium on Vitellogenin and Its mRNA Induction by Estradiol-17$\beta$ in the Primary Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Hwang, Un-Gl;Park, Jin-Il;Shim, Jung-Min;Jung, Chang-Soo;Park, Sung-Yoon
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11a
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    • pp.159-164
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    • 2003
  • Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r=-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

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Hormonal Regulation of Glycerol-Phosphate Acyltransferase Gene Expression (Glycerol-Phosphate Acyltransferase Gene Expression의 호르몬에 의한 조절)

  • 손승렬;신동훈
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.473-477
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    • 1993
  • Both glycerol-phosphate acyltransferase (GPAT) and 7.2 kb mRNAs were present at the highest level in liver. Glycerol-phosphate acyltransferase and 7.2 kb mRNA levels increased dramatically when fasted mice were refed a high carbohydrate diet. In mature 3T3-L1 adipocytes, insulin increased both glycerol-phosphate acyltransferase and 7.2kb mRNA levels 2.6 to 3-fold while dibutyryl cAMP decreased mRNA levels by 50% and 80%, respectively. These results indicate positive regulation by insulin and negative regulation by dibutyryl cAMP of both glycerol-phosphate acyltransferase and 7.2 kb mRNA.

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The Pattern of Cytokine myNA Expression Induced by Polysaccharide from Panax ginseng C. A. Meyer (항암 면역 증강 작용이 있는 인삼 다당체에 의한 cytokine mRNA의 발현)

  • 김기환;이인란;정인성;정희용;윤연숙
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.324-330
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    • 1998
  • We Previously reported that Polysaccharide Isolated from panax ginseng C. A. Meyer, stimulates murine splenocytes to proliferate and to be cytotoxic against a wide range of tumor cells in MHC non-restricted manner:) Therefore, we examined the cytokine mRNA expression induced by the ginseng polysaccharide in this paper. This study demonstrates that the ginseng polysaccharide stimulates Thl type cytosine expression such as IL-2 and IFNY, and macrophage type cytokine expression such as IL-lc and GM-CSF in a dose-dependent manner at different time: IL-2 mRNA was induced at 30 min, IL-la, GM-CSF mRNA at 3 hr, IFNY at 6 hr after the ginseng polysaccharide treatment. In contrast with these, Th2 type cytokine expression such as IL-4 and IL-5 was not induced. The generation of the ginseng polysaccharide-activated killer cells which was induced at the optimal doses of 50 pEyml was neutralized in the presence of anti-lL-2, anti-lFNy, anti-IL-l ${\alpha}$ antibodies, showing the importance of these cytokines produced by the ginseng polysaccharide. In flow cytometry analysis, the blastogenesis of IgM+ cells was induced on day 3 and the number of Thy 1.21 cells, CD4+ and CD8+ cells was increased on day 5. The ginseng polysaccharide also induced blastogenesis of T cells. In conclusion, the ginseng polysaccharide may have considerable antitumor immunotherapeutic modality by stimulating the cytokine production from Thl cells and macrophage and by proliferating lymphocytes.

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Effect of Trolox on Altered Vasoregulatory Gene Expression in Hepatic Ischemia/Reperfusion

  • Eum, Hyun-Ae;Lee, Sun-Mee
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.225-231
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    • 2004
  • This study was designed to investigate the effect of Trolox, a hydrophilic analogue of vitamin E, on the alteration of vasoregulatory gene expression during hepatic ischemia and reperfusion (I/R). Rats were subjected to 60 min of hepatic ischemia in vivo. The rats were treated intravenously with Trolox (2.5 mg/kg) or the vehicle as a control 5 min before reperfusion. Liver samples were obtained 5 h after reperfusion for a RT-PCR analysis on the mRNA for the genes of interest. These mRNA peptides are endothelin-1 (ET -1), potent vasoconstrictor peptide, its receptor $ET_A$ and $ET_B$, vasodilator endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and cyclooxygenase-2 (COX-2). It was seen that serum alanine aminotransferase and lipid peroxi-dation levels were markedly increased after I/R and Trolox significantly suppressed this increase. In contrast, the glutathione concentration decreased in the I/R group, and this decrease was inhibited by Trolox. ET-1 mRNA expression was increased by I/R, an increase which was prevented by Trolox. The mRNA levels for $ET_A$ receptor was significantly decreased, whereas ET$_{B}$ receptor transcript increased in the I/R group. The increase in $ET_A$ was prevented by Trolox. The mRNA levels for iNOS and HO-1 significantly increased in the I/R group and Trolox attenuated this increase. There were no significant differences in eNOS mRNA expression among any of the experimental groups. The mRNA levels for COX-2 and TNF-$\alpha$ significantly increased in I/R group and Trolox also attenuated this increase. Our findings suggest that I/R induces an imbalanced hepatic vasoregulatory gene expression and Trolox ameliorates this change through its free radical scavenging activity.y.

Anti-stress effects of ginseng via down-regulation of tyrosine hydroxylase (TH) and dopamine ${\beta}$-hydroxylase (DBH) gene expression in immobilization-stressed rats and PC12 cells

  • Kim, Yang-Ha;Choi, Eun-Ha;Doo, Mi-Ae;Kim, Joo-Yeon;Kim, Chul-Jin;Kim, Chong-Tai;Kim, In-Hwan
    • Nutrition Research and Practice
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    • v.4 no.4
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    • pp.270-275
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    • 2010
  • Catecholamines are among the first molecules that displayed a kind of response to prolonged or repeated stress. It is well established that long-term stress leads to the induction of catecholamine biosynthetic enzymes such as tyrosine hydroxylase (TH) and dopamine ${\beta}$-hydroxylase (DBH) in adrenal medulla. The aim of the present study was to evaluate the effects of ginseng on TH and DBH mRNA expression. Repeated (2 h daily, 14 days) immobilization stress resulted in a significant increase of TH and DBH mRNA levels in rat adrenal medulla. However, ginseng treatment reversed the stress-induced increase of TH and DBH mRNA expression in the immobilization-stressed rats. Nicotine as a ligand of the nicotinic acetylcholine receptor (nAChR) in adrenal medulla stimulates catecholamine secretion and activates TH and DBH gene expression. Nicotine treatment increased mRNA levels of TH and DBH by 3.3- and 3.1-fold in PC12 cells. The ginseng total saponin exhibited a significant reversal in the nicotine-induced increase of TH and DBH mRNA expression, decreasing the mRNA levels of TH and DBH by 57.2% and 48.9%, respectively in PC12 cells. In conclusion, immobilization stress induced catecholamine biosynthetic enzymes gene expression, while ginseng appeared to restore homeostasis via suppression of TH and DBH gene expression. In part, the regulatory activity in the TH and DBH gene expression of ginseng may account for the anti-stress action produced by ginseng.