• Title/Summary/Keyword: mRNA 발현

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Superoxide Dismutase Gene Expression Induced by Lipopolysaccharide in Alveolar Macrophage of Rat (폐포대식세포에서 내독소 자극에 의한 Superoxide Dismutase 유전자발현의 조절 기전)

  • Park, Kye-Young;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Hyun, In-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.522-534
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    • 1995
  • Background: In the pathogenesis of acute lung injury induced by lipopolysaccharide(LPS), oxygen radiclls are known to be involved in one part. Superoxide dismutase(SOD) protects oxygen radical-induced tissue damage by dismutating superoxide to hydrogen peroxide. In eukaryotic cells, two forms of SOD exist intracellularly as a cytosolic, dimeric copper/zinc-containing SOD(CuZnSOD) and a mitochondrial, tetrameric manganese-containing SOD(MnSOD). But there has been little information about SOD gene expression and its regulation in pulmonary alveolar macrophages(PAMs). The objective of this study is to evaluate the SOD gene expression induced by LPS and its regulation in PAMs of rat. Method: In Sprague-Dawley rats, PAMs obtained by broncholaveolar lavage were purified by adherence to plastic plate. To study the effect of LPS on the SOD gene expression of PAMs, they were stimulated with different doses of LPS($0.01{\mu}g/ml{\sim}10{\mu}g/ml$) and for different intervals(0, 2, 4, 8, 24hrs). Also for evaluating the level of SOD gene regulation actinomycin D(AD) or cycloheximide(CHX) were added respectively. To assess whether LPS altered SOD mRNA stability, the rate of mRNA decay was determined in control group and LPS-treated group. Total cellular RNA extraction by guanidinium thiocyanate/phenolfchlorofonn method and Northern blot analysis by using a $^{32}P$-labelled rat MnSOD and CuZnSOD cDNAs were performed. Results: The expression of mRNA in MnSOD increased dose-dependently, but not in CuZnSOD. MnSOD mRNA expression peaked at 8 hours after LPS treatment. Upregulation of MnSOD mRNA expression induced by LPS was suppressed by adding AD or CHX respectively. MnSOD mRNA stability was not altered by LPS. Conclusion: These findings show that PAMs of rat could be an important source of SOD in response to LPS, and suggest that their MnSOD mRNA expression may be regulated transcriptionally and require de novo protein synthesis without affecting mRNA stability.

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Effects of medicinal herb water extracts on expression of hepatic glucokinase, pyruvate dehydrogenase and acetyl-CoA carboxylase mRNA (한약재 물 추출물이 간세포 Glucokinase, Pyruvate Dehydrogenase, Acetyl-CoA Carboxylase mRNA 발현에 미치는 영향)

  • Kim, Hyun Sook;Kim, Tae Woo;Kim, Dae Jung;Lee, Jae Sung;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.46 no.2
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    • pp.119-125
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    • 2013
  • We studied the anti-diabetic effects of medicinal herb water extracts on expression of hepatic glucokinase (GCK), pyruvate dehydrogenase (PDH), and acetyl-CoA carboxylase (ACC) mRNA. The medicinal herbs used for experiments were Cornus officinalis (CO), Paeonia suffruticosa Andrews (PSA), Discorea japonica Thunb. (DJ), Rehmannia glutinosa (RG), Lycium chinense (LC), and Pyrus pyrifolia (PP). For GCK mRNA expression, CO, RG, and LC water extracts exhibited a more effective activity than other extracts. Cells treated with RG and LC water extracts showed an increase in expression of PDH mRNA to 191% and 124%, respectively, compared to control. Expression of ACC mRNA was significantly higher in LC water extract. These data indicate that CO, RG, and LC water extracts stimulates expression of hepatic GCK, PDH, and ACC mRNA.

Temporal Changes of c-fos, c-jun, and Heat Shock Protein 25 mRNA in Rat Uterus following Estradiol Treatment (Estrogen 처리에 따른 흰쥐 자궁조직내 c-fos, c-jun, hsp25 mRNA 발현 변화)

  • Lee, Young-Ki;Kim, Sung-Rye
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.149-156
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    • 1999
  • Steroid hormone is known to cause the dynamic changes of mammalian uterus during reproductive cycle, which are modulated via hypothalamus-pituitary -gonad reproductive endocrine axis. Although there were so many studies about estrogenic regulation of uterine growth and differentiation. There is little information about the effect of estrogen on the expression of various transcription factors involved in gene expression. Thus the present study was designed to demonstrate E induced expression of c-fos, c-jun, hsp25 mRNA in rat uterus. Employing Northern blot analysis, we studied the temporal expressions of c-fos, c-jun, and hsp25 messenger RNAs (mRNAs) elicited by a single 17beta-estradiol (E) treatment in the uteri of bilaterally ovariectomized adult rats. c-fos, c-jun, and hsp25 mRNA levels were increased and peaked at 3h after E administration, and then c-fos and c-jun mRNA levels were rapidly decreased to basal control level while, increased hsp25 mRNA levels were sustained till 12h post E treatment. To test the estrogenic effect on the increase of c-fos, c-jun, and hsp25 mRNA levels, we also examined the effects of antiestrogen (tamoxifen). Pretreatment with tamoxifen effectively blocked the E-induced increase of c-fos, c-jun, and hsp25 mRNA levels at 3h post E treatment. Present results suggest that transient increase of c-fos and c-jun protooncogene mRNA at the early time and simultaneous expression of hsp25 mRNA contribute to the response of uterine tissues to E in adult female rats.

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Exploring Cancer-Specific microRNA-mRNA Interactions by Evolutionary Layered Hypernetwork Models (진화연산 기반 계층적 하이퍼네트워크 모델에 의한 암 특이적 microRNA-mRNA 상호작용 탐색)

  • Kim, Soo-Jin;Ha, Jung-Woo;Zhang, Byoung-Tak
    • Journal of KIISE:Computing Practices and Letters
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    • v.16 no.10
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    • pp.980-984
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    • 2010
  • Exploring microRNA (miRNA) and mRNA regulatory interactions may give new insights into diverse biological phenomena. Recently, miRNAs have been discovered as important regulators that play a major role in various cellular processes. Therefore, it is essential to identify functional interactions between miRNAs and mRNAs for understanding the context- dependent activities of miRNAs in complex biological systems. While elucidating complex miRNA-mRNA interactions has been studied with experimental and computational approaches, it is still difficult to infer miRNA-mRNA regulatory modules. Here we present a novel method, termed layered hypernetworks (LHNs), for identifying functional miRNA-mRNA interactions from heterogeneous expression data. In experiments, we apply the LHN model to miRNA and mRNA expression profiles on multiple cancers. The proposed method identifies cancer-specific miRNA-mRNA interactions. We show the biological significance of the discovered miRNA- mRNA interactions.

Effect of 4-Nonylphenol on the Gene Expression of Retinol-Binding Protein in the Rockfish, Sebastes schlegeli (조피볼락(Sebastes schlegeli)의 Retinol-Binding Protein의 유전자 발현에 미치는 4-Nonylphenol의 영향)

  • Cho, Hyung-Koo;Jung, Jee-Hyun;Lee, Je-Yong;Kim, Myung-Hee;Han, Chang-Hee
    • Development and Reproduction
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    • v.10 no.3
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    • pp.177-184
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    • 2006
  • Retinol-binding protein(RBP) plays an important role in the specific transport of retinol to target cells through the blood stream in higher vertebrates. In order to clarify the effects of 4-nonylphenol(4-NP) on RBP mRNA expression in the rockfish, Sebastes schlegeli which is common in coastal waters of Korea and commercially important species, the cDNA library was constructed from the liver, and a partial fragment of the RBP gene was cloned. The deduced amino acid sequence from the RBP mRNA showed a high homology to the amino acid sequence from Sparus aurata(80%), Oncorhynchus mykiss(72%) or Anguilla anguilla(78%). Effects of 4-NP on RBP and vitellogenin(VTG) mRNA expression level in rockfish were examined by the northern blot analysis. In female and male rockfish injected with 4-NP(10 mg/kg BW, lower dose), there was no changes in the levels of VTG mRNA expression in the liver. The RBP mRNA levels, however, decreased at 48 hours after the injection in male. In the rockfish injected with 4-NP(25 mg/kg BW, higher dose), the level of VTG mRNA expression increased after 24 hours, regardless of sex. The level of RBP mRNA expression decreased at 48 hours after the injection in both sexes. These data indicate that estrogenic mimics such as 4-NP exhibit a contrasting effect on RBP and VTG gene expression in rockfish.

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The Mechanism of LDL Receptor Up-regulation by Ginsenoside-Rb2 in HepG2 Cultured under Enriched Cholesterol Condition (고콜레스테롤 조건하에 배양된 HepG2에서의 ginsenoside-Rb2에 의한 LDL receptor 억제 완화 기전)

  • Lim, G-Rewo;Lee, Hyun-Il;Kim, Eun-Ju;Ro, Young-Tae;Noh, Yun-Hee;Koo, Ja-Hyun
    • Journal of Ginseng Research
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    • v.28 no.2
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    • pp.87-93
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    • 2004
  • The effect of ginsenoside-Rb2, one of a major pharmacological component of Panax ginseng C.A. Meyer, on low density lipoprotein (LDL) receptor expression was investigated and compared with hypocholesterolemic drug lovastatin. In HepG2 cell, exogenous cholesterol decreased LDL receptor mRNA expression, but ginsenoside-Rb2 recovered this reduction of LDL receptor mRNA up to normal expression level. Lovastatin also increased LDL receptor mRNA expression as similar as ginsenoside-Rb2 did. The reduction of sterol regulatory element binding protein (SREBP) transcription by exogenous cholesterol was also similarly recovered by ginsenoside-Rb2 and lovastatin addition. Compound K, a metabolite of ginsenoside-Rb2 and -Rb1 by human intestinal bacteria also increased the SREBP mRNA expression in cholesterol-enriched condition. Ginsenoside-Rb2 seems to up-regulate LDL receptor mRNA expression through the induction of de novo SREBP transcription. Therefore, increased expression of SREBP mRNA by ginsenoside-Rb2 elevated the LDL receptor mRNA expression in HepG2 cells, and these inductions possibly drop the plasma cholesterol level in hypercholesterolemia patients, in vivo, as likely in case of lovastatin.

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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Differential Expression of Glycoprotein Hormones in Equine Placenta and Pituitary (말 태반과 뇌하수체에서 당단백질 호르몬의 특이적인 발현)

  • Min, Kwan-Sik
    • Development and Reproduction
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    • v.4 no.1
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    • pp.87-93
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    • 2000
  • Equine chorionic gonadotropin (eCG) consists of highly glycosylated noncovalently linked $\alpha$- and $\beta$-subunits and belongs to the glycoprotein hormone family that includes lutropin (LH), follitropin (FSH), and thyrotropin (FSH). eCG is a unique member of the gonadotropin family because it elicits response characteristics of both FSH and LH in other species than the hone. eCG is synthesized and secreted by trophoblastic cells of the endometial cups between 40 and 130 days of gestation. In the present study, mRNA expression ratio of eCG, eLH and eFSH $\alpha$- and $\beta$-subunints was investigated in the placenta and pituitary. mRNA was extracted from equine placenta on day 70 of gestation and from pituitary of male horse (27 month-old). When the expression of both subunit mRNAs of eCG in the equine placenta was compared by Northern blotting, the expression of the $\beta$ -subunit mRNA was relatively greater than that of the $\alpha$-subunit. And mRNA expression of $\alpha$-, LH $\beta$- and FSH $\beta$-subunits was analysed in the equine pituitary. An $\alpha$-subunit was revealed with a size of approximately 0.8 kb. FSH $\beta$-subunit mRNA also was detected out 1.8 kb. It is the same size of the FSH $\beta$ -subunit mRNA cloned. The intensity of $\alpha$-subunit mRNA was greater than that of the $\beta$-subunit suggesting that the expression of $\alpha$ -subunit was dominant in the equine anterior pituitary. Thus, the subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$- and $\beta$-subunits in the equine placenta and pituitary.

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Effects of Medicinal Plant Water Extracts on Expression of Anti-diabetic Enzymes mRNA (약용식물 물 추출물이 항당뇨 효소의 유전자 발현에 미치는 영향)

  • Kim, Hyun Sook;Kim, Tae Woo;Kim, Dae Jung;Kim, Kyoung Kon;Choe, Myeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1008-1014
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    • 2013
  • This study evaluated the anti-diabetic effects of medicinal plant water extracts on expression of hepatic glucokinase (GCK), pyruvate dehydrogenase (PDH) and acetyl-CoA carboxylase (ACC) mRNA. GCK, PDH and ACC mRNA expression levels were measured by RT-PCR. The medicinal plants used in our study were Cordyceps militaris (CM), Perilla sikokiana (PS), Salvia miltiorrhiza Bunge (SMB), Panax notoginseng (PN) and Angelica utilis Makino (AUM). We found that GCK mRNA expression was increased to about 181% at the 250 ppm of CM water extract. Furthermore, we also found that CM and AUM water extracts stimulated PDH mRNA expression level related to glucose metabolism, however, PS, SMB and PN did not stimulate PDH mRNA expression as expected. Expression of ACC mRNA was also significantly higher in both CM and AUM water extracts. Overall, the results of our study suggest that CM and AUM water extracts stimulate expression of hepatic GCK, PDH and ACC mRNA.

Effects of Relative Lysyl Oxidase and Hydrogen Peroxide on Odontoblastic Differentiation (인간치수세포 분화과정에서 과산화수소에 대한 Lysyl Oxidase의 역할)

  • Lee, Hwa-Jeong
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.321-329
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    • 2013
  • Although it has been reported that lysyl oxidase (LOX) is involved in odontoblastic differentiation, the role of LOX on odontoblastic differentiation by hydrogen peroxide ($H_2O_2$) have not been clarified. In the present study, we investigated whether $H_2O_2$, reactive oxygen species (ROS), is modulated the messenger RNA (mRNA) expression and activity of LOX during odontoblastic differentiation of human dental pulp (HDP) cells. The mRNA expression was quantified by reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and LOX enzyme activity was measured by high sensitive fluorescent assay. Expression of the odontoblastic differentiation marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX and LOXL. The $H_2O_2$-induced mRNA expression of LOX family was significant reduction of LOX, LOXL, and LOXL3 mRNA levels in HDP cells. LOX enzyme activity was increased at $H_2O_2$ 0.3 mM for 24 hours. The mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) was inhibited by LOX- and LOXL-specific siRNAs whereas the mRNA expression of dentin matrix protein1 (DMP1), and dentin sialophosphoprotein (DSPP) was inhibited by LOX-specific siRNA. In LOX enzyme activity, siRNA-induced knockdown of both LOX and LOXL inhibited the total amine oxidase activity in HDP cells, as in the case of mRNA expression. In conclusion, the essential role of $H_2O_2$ on odontoblastic differentiation suggests that its regulation by LOX may have pharmacologic importance in HDP cells.