• Title/Summary/Keyword: mRNA activation

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Regulation of Immediate Early Gene Expression by Glutamate Receptor Activation in C6 Rat Glioma Cells

  • Lee, Jin-Koo;Kim, Yung-Hi;Choi, Seong-Soo;Suh, Hong-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.19-25
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    • 2001
  • We have studied the effects of excitatory amino acids on the expression of the c-fos and c-jun mRNA in rat C6 glioma cells. The glutamate, $N-methyl-_D-aspartate$ (NMDA), and kainic acid (KA) increased c-fos mRNA level in a concentration-dependent manner. However, they did not affect c-jun mRNA level. In addition, forskolin and phorbol 12-myristate 13-acetate (PMA) increased c-fos mRNA level. Furthermore, PMA increased c-jun mRNA level whereas forskolin downregulated c-jun mRNA level. The glutamate, NMDA and KA, at a concentration of 0.25 mM, did not affect the basal c-fos and c-jun mRNA levels, and also did not affect forskolin- and PMA-induced responses. Furthermore, both forskolin and PMA itself increased the phosphorylation of ERK (extracellular signal regulated kinase) and CREB (cyclicAMP responsible element binding protein) proteins. The KA, NMDA, and glutamate did not affect forskolin- induced increase of ERK and CREB phosphorylation. The KA decreased PMA-induced increase of phosphorylation of ERK and CREB proteins, whereas glutamate and NMDA did not affect the phosphorylation of ERK and CREB proteins induced by PMA. These findings suggest that, in C6 glioma cells, c-fos mRNA induction induced by EAAs is not mediated by phosphorylation of ERK and CREB proteins.

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Induction of Nitric Oxide and Cytokines in Macrophages by Codonopsis lanceolata (대식세포에서 산더덕에 의한 NO 생성 및 싸이토카인 유도효과)

  • So, Mi-Sun;Lee, Jin-Sil;Yi, Seh-Yoon
    • Korean Journal of Food Science and Technology
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    • v.36 no.6
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    • pp.986-990
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    • 2004
  • The immunomodulatory effect of Codonopsis lanceolata based on the production of cytokines and the activation of macrophage was studied. The mRNA expression of nitric oxide synthase (iNOS) was gradually induced after 24 hr treatment of Codonopsis lanceolata, and NO production was a maximum after 24 hr treatment with 1 mg/mL. RAW 264.7 cell on in vitro treatment with Codonopsis lanceolata induced mRNA of cytokines such as interleukin-1(IL-1)${\beta}$, interleukin-6(IL-6), tumor necrosis $factor(TNF)-{\alpha}\;and\;interferon(IFN)-{\gamma}$; $IL-1{\beta}$ and IL-6 mRNA were gradually induced up to 24 hr, $TNF-{\alpha}\;mRNA$ was regularly induced up to 24 hr, and $IFN-{\gamma}\;mRNA$ level was a maximum within 1 hr. These results suggest that Codonopsis lanceolata exerts as an effective immunomodulator and enhances antitumor activity of macrophages.

Ginsenoside Rb2 Upregulates the Low Density Lipoprotein Receptor Gene Expression through the Activation of the Sterol Regulated Element Binding Protein Maturation in HepG2 Cells

  • Lim, Grewo;Lee, Hyunil;Kim, Eun-Ju;Noh, Yun-Hee;Ro, Youngtae;Koo, Ja-Hyun
    • Journal of Ginseng Research
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    • v.29 no.4
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    • pp.159-166
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    • 2005
  • Ginsenosides, a group of Panax ginseng saponins, exert the lowering effects of plasma cholesterol levels in animals. We had reported earlier that ginsenoside Rb2 upregulate low-density lipoprotein receptor (LDLR) expression via a mechanism that is dependent of the activation of sterol response element binding protein 2 (SREBP-2) expression. This study was conducted to determine the effects of ginsenoside Rb2 on the expression of the hepatic LDLR expression at cellular levels using HepG2 cells, and to evaluate whether the sterol response element binding protein 1 (SREBP-l) was involved in the regulation of LDLR expression. Incubation of HepG2 cells in serum-free medium supplemented with cholesterol $(10{\mu}g/ml)$ for 8 hours decreased the mRNAs of LDLR mRNA by $12\%$ and SREBP-l mRNA by $35\%$. Ginsenoside Rb2 antagonized the repressive effects of cholesterol and increased both LDLR and SREBP-l mRNA expression to 1.5- and 2-fold, respectively. Furthermore, Western blot and confocal microscopic analyses with SREBP-l polyclonal antibody revealed that ginsenoside Rb2 enhanced the maturation of the SREBP-1 from the inactive precursor form in ER membrane to the active transcription factor form in nucleus. These results suggest that ginsenoside Rb2 upregulates LDLR expression via a mechanism that is dependent of the activation of not only SREBP-2 expression, but also SREBP-1 expression and maturation, and also indicate that the pharmacological value of ginsenoside Rb2 may be distinguished from that of lovastatin which is reported that it upregulate LDLR through SREBP-2 only, not through SREBP-1.

RNA Metabolism in T Lymphocytes

  • Jin Ouk Choi;Jeong Hyeon Ham;Soo Seok Hwang
    • IMMUNE NETWORK
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    • v.22 no.5
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    • pp.39.1-39.18
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    • 2022
  • RNA metabolism plays a central role in regulating of T cell-mediated immunity. RNA processing, modifications, and regulations of RNA decay influence the tight and rapid regulation of gene expression during T cell phase transition. Thymic selection, quiescence maintenance, activation, differentiation, and effector functions of T cells are dependent on selective RNA modulations. Recent technical improvements have unveiled the complex crosstalk between RNAs and T cells. Moreover, resting T cells contain large amounts of untranslated mRNAs, implying that the regulation of RNA metabolism might be a key step in controlling gene expression. Considering the immunological significance of T cells for disease treatment, an understanding of RNA metabolism in T cells could provide new directions in harnessing T cells for therapeutic implications.

Expression of osteopontin and this role in hepatic stellate cell motility and wound healing migration

  • Lee, Sung-Hee;zhe, Zhao-Yu;Park, Eun-jeon;Min, Song-Hye;Hwan, Sohn-Dong
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.142.2-142.2
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    • 2003
  • The activation of the hepatic stellate cell (HSC) is a key step in liver fibrogenesis. We investigated the changes of global gene expression during activation in hepatic stellate cells using a rat cDNA microarray with 5, 000 sequence-verified clones. We identified osteopontin (OPN), a secreted matrix protein, as one of the upregulated factors. Northern analysis showed OPN mRNA was increasingly expressed during progressive activation of cultured rat HSCs and in models of experimental liver fibrosis. (omitted)

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Endoplasmic Reticulum (ER) Stress and Apoptosis in Parthenogenetic Porcine Embryos following Different Combination of Activation Methods

  • Park, Hye-Bin;Park, Yeo-Reum;Lee, Hwa-Yeon;Bae, Hyo-Kyung;Lee, Seunghyung;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Journal of Embryo Transfer
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    • v.32 no.1
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    • pp.25-31
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    • 2017
  • This study was conducted to investigate the effect of activation method on the endoplasmic reticulum (ER) stress induction, apoptosis and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by four activation methods; 1) electric stimulus (ES) (E), 2) $ES+10{\mu}M$ Ca-ionophore (A23187) treatment (EC), 3) ES+2 mM 6-dimethylaminopurine (6-DMAP) treatment (ED), or 4) ES+A23187 and 6-DMAP treatments (ECD). Parthenogenetic embryos were sampled to analyze x-box binding protein 1 (Xbp1) mRNA, ER stress-associated genes and apoptosis genes at 3 h after ES and the 1-cell and blastocyst stages. In the EC group, the band intensity of spliced Xbp1 (Xbp1s) mRNA was higher than those of the other groups at the 3 h and 1-cell stage, and higher than that of the E group at the blastocyst stage. Four ER stress-associated genes were expressed at the highest level in the EC group and weakly expressed in the ED group at 3 h after activation. However, most of the genes were highly expressed at the 1-cell and blastocyst stages with some variation in the EC and ECD groups. Expression of Bcl-2-associated X protein (Bax) and caspase-3 mRNA was significantly higher in the EC group than in the other groups at all development stages. The developmental rates to the blastocyst stage were higher in the ED and ECD groups than in the E and EC groups. These results suggest that the intracellular ER stress of parthenogenetic porcine embryos is affected by the activation method and subsequently lead to the apoptosis of embryos.

Effects of Fucoidan on Nitric Oxide Production and Activator Protein-1 Activation in Lipopolysaccharide-Stimulated Porcine Peripheral Blood Mononuclear Cells (LPS로 자극한 돼지 말초혈액 단핵구세포의 Nitric Oxide (NO) 생산 및 Activator Protein-1 (AP-1) 활성화에 있어 Fucoidan의 효과)

  • Park, Jongchan;Ahn, Changhwan;Kang, Byeong-Teck;Kang, Ji-Houn;Jeung, Eui-Bae;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.32 no.4
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    • pp.289-294
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    • 2015
  • Fucoidan which is sulfated polysaccharide extracted from brown seaweed has a wide variety of internal biological activities. The objectives of this study were to examine the effect of fucoidan on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to investigate whether this effect is involved in the expression of inducible nitric oxide synthase (iNOS) and the activation of activator portein-1 (AP-1). The levels of NO production and AP-1 activity in the culture supernatants from porcine PBMCs were measured by the enzyme-linked immunosorbent assay and the levels of iNOS and AP-1 mRNA were determined by real time polymerase chain reaction. Fucoidan in LPS-naïve PBMCs has no effects on the production of NO and activity of AP-1. Expressions of iNOS and AP-1 mRNA in LPS-naïve PBMCs were also not affected by treatment of fucoidan. However, NO production, AP-1 activity and expressions of iNOS and AP-1 mRNA were dramatically increased in PBMCs stimulated with LPS. Enhancing effects of NO production and AP-1 activity in PBMCs induced by LPS were reduced by addition of fucoidan. Fucoidan also inhibited an increase in expressions of iNOS and AP-1 mRNA in LPS-stimulated PBMCs. These results suggested that fucoidan exerts anti-inflammatory effect by down-regulating production of NO via suppressing expression of iNOS and activity of AP-1 in LPS-stimulated porcine PBMCs.

Suppressive Effects of GST on Cytokine-induced Activation of Human Fibroblast-like Sinoviocytes (가미사물탕(GST)의 사이토카인으로 유도된 인간 섬유아세포양 활막 세포 활성화 저해 작용)

  • Park, Jee-Young;Jin, Mi-Rim;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.14 no.2
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    • pp.45-54
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    • 2005
  • GST, an extract from 16 herbs, has been formulated and prescribed for the treatment of human rheumatoid arthritis(hRA) for many years. The present study was done to investigate whether GST has suppressive effects on activation of fibroblast-like sinoviocytes isolated from an RA patient. In tumor necrosis factor-a(TNF-a)/interleukin-1b(IL-1b) treated human sinoviocytes, The mRNA expression of molecular indicators related to pathologic changes of the sinoviocytes were examined using quantitative real-time PCR. The treatment of GST($100\;{\mu}g/ml$) suppressed the expression of proinflammatory cytokines and chemokines such as TNF-a, IL-1b, IL-6 and IL-8 compared with the control. The mRNA level of intracellular adhesion molecule-1(ICAM-1) which is known to increase in the activated sinoviocytes of RA patients, was slightly decreased by GST. The expression of NOS-II was considerably reduced, which was accompanied by a decrease in the production of nitric oxide(NO). In addition, GST considerably increased the mRNA levels of tissue inhibitors of matrix metalloproteinase-1(TIMP-1), while those of matrix metalloproteinase-3(MMP-3) were decreased. Taken together, these data suggested that GST might suppress the activation of sinoviocytes in hRA.

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Inhibitory Effects of GC, an Extract from Herbs, on $TNF-{\alpha}/IL-1{\beta}$-induced Activation of Human Fibroblast-like Sinoviocytes (계혈등복합방(GC)의 $TNF-\alpha$$IL-1{\beta}$로 유도된 인간 섬유아세포양 활막 세포 활성화 억제 작용)

  • Jang Kwang-Ho;Jin Mi-Rim;Park Hee-Ok;Kim Dong-Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1225-1232
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    • 2005
  • Based on traditional medicine theories, GC, an extract from 5 herbs, has been formulated and prescribed for the treatment of human rheumatoid arthritis(hRA) for many years. The present studies was done to investigate whether GC has inhibitory effects on activation of fibroblast-like sinoviocytes isolated from a RA patient. In tumor necrosis factor-${\alpha}(TNF-{\alpha}$)/ interleukin-IL-$1{\beta}$(IL-$1{\beta}$) treated human sinoviocytes, the mRNA expression of molecular indicators related to pathologic changes of the sinoviocytes were examined using quantitative real-time PCR. The treatment of GC($10{\mu}g/ml$) significantly suppressed the expression of proinflammatory cytokines and chemokines such as $TNF-{\alpha}$, IL-6 and IL-8 compared with the control, but not $IL-1{\beta}$, The mRNA level of intracellular adhesion molecule-1 (ICAM-1) which is known to increase in the activated sinoviocytes of RA patients, was slightly decreased by GC. The expression of NOS-II was considerably reduced, which was accompanied by a decrease in the production of nitric oxide(NO). Furthermore, GC dramatically raised the mRNA levels of tissue inhibitors of matrix metalloproteinase-1 (TIMP-1), while those of matrix metalloproteinase-3 were significantly lowered. Taken together, these data suggested that GC might suppress the activation of sinoviocytes in hRA.

Chunghyuldan Downregulates the Activation of Transcription Factors NF - kB and AP-1 of BV-2 Cells Induced by Lipopolysaccharide

  • WEE Sung-SooK;BAE Eun-Ah;PARK Jin-Sun;KIM Hee-Sun;CHo Hee Jae;Ryu Jong-Hoon;KIM Dong-Hyun
    • Biomolecules & Therapeutics
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    • v.13 no.4
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    • pp.214-219
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    • 2005
  • Chunghyuldan (Qingxuedan in Chinese) (CHD) has been used for patients with atherosclerosis and brain ischemia in Korea. To evaluate antiischemic activity of CHD, its antiinflammatory effect in lipopolysaccharide-induced BV-2 cells was investigated. CHD potently inhibited nitric oxide (NO) production in LPS-induced BV-2 cells with an $IC_{50}$ value of 4.8${\mu}g/ml$. CHD did not only inhibit mRNA and protein expression levels of inducible NO synthase and cyclooxygenase-2 in LPS-induced BV-2 cells, but also repressed mRNA expression levels of proinflammatory cytokines IL-l$\beta$ and TNF-$\alpha$. CHD also downregulated the activation of NF-kB and AP-l transcription factors induced by LPS. These results suggest that CHD may improve inflammatory brain ischemia by the downregulation the activation of NF-kB and AP-l transcription factors.