• Title/Summary/Keyword: mRNA 발현

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Ethanol Extract of Hippophae Rhamnoides L. Leaves Inhibits Adipogenesis through AMP-activated protein kinase (AMPK) Activation in 3T3-L1 Preadipocytes (비타민나무 잎 에탄올추출물의 AMPK 활성화를 통한 3T3-L1 지방전구세포의 adipogenesis 억제효과)

  • Jeong, Hyeon Ju;Park, Ju Hee;Kim, Myong-Jo
    • Korean Journal of Plant Resources
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    • v.28 no.5
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    • pp.582-590
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    • 2015
  • In the present study, we investigated the effect of 70% EtOH extract from Hippophae Rhamnoides L. leaves (HRL) on the anti-obesity effect in 3T3-L1 cells. The effects of HRL on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. In addition, we examined the gene expression levels by using RT-PCR and western blot. The results of this analysis showed that 100 ㎍/㎖ HRL significantly increased the inhibition of lipid accumulation by 82.25%; significantly decreased the mRNA expression of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα), and fatty acid synthase (FAS) in 3T3-L1 cells as well as the stimulated protein expression of AMP-activated protein kinase (AMPK); and suppressed the expression level of PPARγ. These results suggest that HRL can prevent adipogenesis through activation of AMPKα and inhibition of adipogenesis transcription factors.

A Study on the Expression of Thymosin-β4 and c-Myc mRNA in the Model of liver cirrhosis with fibrosis (섬유화 진행 간경변 조직 모델에서 Thymosin β4와 C-myc mRNA 융합 발현 연구)

  • Kim, Jean-Soo;Park, Un-Kyu
    • Journal of the Korea Convergence Society
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    • v.10 no.6
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    • pp.65-71
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    • 2019
  • The propose of this study has been conducted to examine expression of c-Myc and Thymosin-${\beta}4$ in liver cirrhosis model from liver fibrosis and For the method of study, the experiment was conducted in 2 groups; liver cirrhosis model experiment group due to liver fibrosis and control group with distilled water. This study outcome showed that liver cirrhosis model experiment group had significantly higher expression of c-Myc and Thymosin-${\beta}4$. with changes to hepatic tissue of special staining and electron microscopy. In conclusion, in clinical tests regarding liver function, molecular evaluation of c-Myc and Thymosin-${\beta}4$ and their expression along with serological change and histological assessment can be utilized as a reference for diagnosing liver disease for prevention and diagnosis of the disease, Based on this research in the future, we will carry out an in-depth study by adding the types of experimental groups and related genes.

Export of Human Proinsulin in E. coli : High Export of Proinsulin Fusion Protein but not of Proinsulin Itself (대장균에서 인체 프로인슐린의 분비 발현 : 프로인슐린 융합체의 고분비 발현과 프로인슐린의 저분비 발현)

  • Yup Kang
    • KSBB Journal
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    • v.11 no.2
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    • pp.165-172
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    • 1996
  • To obtain a correctly folded human proinsulin, export of proinsulin using Staphylococcal protein A signal sequence-mediated secretion pathway has been attempted in E.coli. A secretion operon for proinsulin was constructed by consecutively connecting T7 promoter, SPA ribosome binding site, SPA signal sequence gene, and human proinsulin gene. Little immunoreactive proinsulin was detected in the periplasmic space and. culture medium, and not even in cytoplasmic space. The qualitative analysis of transcribed proinsulin mRNA and the in vitro transcription/translation experiment suggests that the negligible level of proinsulin export appears to be due to intracellular degradation of proinsulin, rather than due to the blockage during translocation. However, expression of proinsulin fusion protein such as MBP-proinsulin could dramatically increase export of proinsulin in E.coli.

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Pro-inflammatory Cytokine Expression Through NF-${\kappa}B/I{\kappa}B$ Pathway in Lung Epithelial Cells (폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로에 의한 염증매개 사이토카인의 발현)

  • Park, Gye-Young;Lee, Seung-Hee;HwangBo, Bin;Yim, Jae-Joon;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.332-342
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    • 2000
  • Background : The importance of pro-inflammatory cytokines, especially tumor necrosis factor $\alpha$ (INF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$), have been extensively documented in the generation of inflammatory lung disease. Lung epithelial cells are also actively involved in initiating and maintaining inflammation by producing pro-inflammatory mediators. Understanding the mechanism of pro-inflammatory cytokine expression in lung epithelial cells is crucial to the development of new therapeutic modalities for inflammatory lung disease. Transcription of most pro-inflammatory cytokines is dependent on the activation of NF-${\kappa}B$. However, the relationship between pro-inflammatory cytokine expression and NF-${\kappa}B/I{\kappa}B$ pathway in lung epithelial cells is not clear. Methods : BEAS-2B, A549, Na-H157, NCI-H719 cells were stimulated with IL-$1{\beta}$ or TNF-$\alpha$ at various times, and then IL-8 and TNF-$\alpha$mRNA expressions were assayed by Northern blot analysis. IL-$1{\beta}$ or TNF-$\alpha$-induced NF-${\kappa}B$ activation was assessed by the nuclear translocation of p65 NF-${\kappa}B$ subunit. The degradation of $I{\kappa}B{\alpha}$ and $I{\kappa}B{\beta}$ by IL-$1{\beta}$ or TNF-$\alpha$stimulation was assayed by Western blot analysis. The phosphorylation of $I{\kappa}B{\alpha}$ was evaluated by Western blot analysis after pre-treating cells with proteasome inhibitor followed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation. The basal level of IKK $\alpha$ expression was evaluated by Western blot analysis. Results: $I{\kappa}B{\alpha}$ and $I{\kappa}B{\alpha}$ was rapidly degraded after 5 minutes of incubation with IL-$1{\beta}$ or TNF-$\alpha$ in BEAS-2B, A549, and NCI-H157 cells. The activation of NF-${\kappa}B{\alpha}$ and the induction of IL-8 and TNF-$\alpha$ mRNA expression were observed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in these cells. In contrast, neither the changes in NF-${\kappa}B/I{\kappa}B$ pathway nor IL-8 and TNF-$\alpha$mRNA expression was induced by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in NCI-H719 cells. IL-$1{\beta}$ and TNF-$\alpha$-induced $I{\kappa}B$ phosphorylation was observed in BEAS-2B, A549, and NCI-H157 cells, but not in NCI-H719 cells. The basal level of IKK$\alpha$ expression was not different between cell. Conclusion : NF-${\kappa}B/I{\kappa}B$ pathway plays an important role in the expression of pro-inflammatory cytokine in most lung epithelial cells. The absence of the effect on NF-${\kappa}B/I{\kappa}B$ pathway in NCI-H719 cells sæms to be due to the defect in the intracellular signal transduction pathway upstream to IKK.

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Changes in enzyme activity and expression of DHFR of Toxoplumc gondii by antifolates (Antifolate 약제에 대한 톡소포자충의 DHFR 효소활성 및 유전자 발현의 변화)

  • 백은정;남호우
    • Parasites, Hosts and Diseases
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    • v.36 no.3
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    • pp.191-198
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    • 1998
  • The responses to antifolales of ToxopLasmc Bondii were investigated by measuring the dihydrorolate redLlctase (DHFR) activity. quantity of DHFR mRNA, and single-strand conformational polymorphism (SSCP) pattern. Pyrimethamine (PYM) and methotrexate (MTX) were tested ds anlifolates. When T. gondii was treated wish PYM, the viability was decreased by the increasing concentration of PYM. DHFR activity tended to increase as the passage proceeded. and the quantity of mRNA expressed was also increased according to passages. The viability of 7. gonnii was decreased by the increasing concentration of MTX, but it was maintained over 40% up to $100{\;}{\mu\textrm{m}}$ MTX. DHFR activity was 77.4% in the 1st passage ($1{\;}{\mu\textrm{m}}$). 82.2% in the 4th passage ($10{\;}{\mu\textrm{m}}$), and 141.3% in the 7th passage ($100{\;}{\mu\textrm{m}}$) But no changes were detected in SSCP pattern of T gondii rxposvd to FYM and MTX. both. These results suggested that the response of T gondii to FYN was rofulalcd by transrriptional level and that, in MTX. the viability of T. gonnii was derived from increasing DHFK activity.

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Smad6 Gene and Suppression of Radiation-Induced Apoptosis by Genistein in K562 Cells (K562 세포주에서 Genistein에 의해 억제되는 Radiation-induced Apoptosis의 조절 유전자)

  • Jeong, Soo-Jin;Jin, Young-Hee;Yoo, Yeo-Jin;Do, Chang-Ho;Jeong, Min-Ho;Huh, Gi-Yeong;Bae, Hye-Ran;Yang, Kwang-Mo;Moon, Chang-Woo;Oh, Sin-Geun;Hur, Won-Joo;Lee, Hyung-Sik
    • Radiation Oncology Journal
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    • v.19 no.3
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    • pp.245-251
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    • 2001
  • Prupose : The genes involved on the suppression or radiation-induced apoptosis by genistein in K562 leukemia cell line was investigated. Materials and methods : K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For X-ray irradiation and drug treatment, cultures were prepared at $2\times10^5\;cells/mL$. The cells were irradiated with 10 Gy (Clinac 1800C, Varian, USA), Stock solutions of herbimycin A (HMA, Calbiochem, UK) and genistein (Calbiochem, UK) were prepared in dimethylsulfoxide (DMSO, Sigma, UK). After incubation at $37^{\circ}C$ for 24 h, PCR-select cDNA subtractive hybridization, dot hybridization, DNA sequencing and Northern hybridization were examined. Results : Smad6 gene was identified from the differentially expressed genes in K562 cells incubated with genistein which had been selected by PCR-select cDNA subtractive hybridization. The mRNA expression of Smad6 in K562 cells incubated with genistein was also higher than control group by Northern hybridization analysis. Conclusion : We have shown that Smad6 involved on the suppression of radiation-induced apoptosis by genistein in K562 leukemia cell line. It is plausible that the relationship between Smad6 and the suppression of radiation-induced apoptosis is essential for treatment development based on molecular targeting designed to modify radiation-induced apoptosis.

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Physiological Studies on Acute Water-temperature Stress of Juvenile Abalone, Haliotis discus hannai (급격한 수온 스트레스에 따른 전복, Haliotis discus hannai 치패의 생리학적 연구)

  • Kim Tae-Hyung;Yang Moon-Hyu;Choe Mi-Kyung;Han Seok-Jung;Yeo In-Kyu
    • Journal of Aquaculture
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    • v.18 no.1
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    • pp.7-12
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    • 2005
  • This study was conducted to investigate antioxidant enzyme activity (catalase and superoxide dismutase) and Heat Shock Protein 70 (HSP70) mRNA variation in hepatopancreas of abalone (Haliotis discus hannai) cultured under several acute water temperatures. Abalones were cultured at 10, 15, 20, 25 and $30^{\circ}C$, for 0, 6, 12, 24 and 48 hours, respectively. The HSP70 mRNA expression in hepatopancreas was more increased at $30^{\circ}C$ compared to those at 10. 15, 20 (control) and $25^{\circ}C$. The superoxide dismutase (SOD) activity was increased in hepato-pancreas at all water temperature conditions compared to the control ($20^{\circ}C$). The SOD activity at high water temperature (25 and $30^{\circ}C$) tended to be increased after 12 hours, and was increased immediately after exposure to low water temperature (10 and $15^{\circ}C$). and then was recovered to starting level after the increase. Also, catalase (CAT) activity in hepatopancreas was increased in all the groups except for at $10^{\circ}C$ than the control ($20^{\circ}C$). Survival rate of abalone was $100\%$ at 10, 15, 20 and $25^{\circ}C$, but $92\%$ at $30^{\circ}C$. Thus, according to our study, when abalone is appeared at $20^{\circ}C$, defense mechanism against stress at low water temperature can be accelerated to be stabilized at about $5^{\circ}C$. In the case of exposure of abalone to high water temperature, antioxidant enzyme and HSP70 expression were increased due to elevated physiological stimulation factor, such as temperature.

Effect of Submerged Culture of Ceriporia lacerata Mycelium on Insulin Signaling Pathway in 3T3-L1 Cell (Ceriporia lacerata 균사체 배양물이 3T3-L1 세포에서 인슐린 신호 전달에 미치는 영향)

  • Shin, Eun Ji;Kim, Ji-Eun;Kim, Ji-Hye;Park, Yong Man;Yoon, Sung Kyoon;Jang, Byeong-Churl;Lee, Sam-Pin;Kim, Byoung-Cheon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.325-330
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    • 2016
  • In this study, we evaluated the antidiabetic effect of submerged culture of Ceriporia lacerata mycelium (CL01) on glucose uptake and the expression of mRNA and protein of major signal markers of insulin signaling pathway in 3T3-L1 adipocytes. After 3T3-L1 adipocytes were pre-treated by CL01 (0, 2, 10 mg/ml) for 8 hours, followed with treatment of insulin, the glucose uptake levels significantly increased by more 55.1%, 94.4% than negative control respectively (p<0.01, 0.001) in a dose-dependent manner. However, in case of CL01 pre-treatment without insulin, the glucose uptake did not increase compared with insulin-treated 3T3-L1. Also we demonstrated that the protein expression levels of pIR β, pAkt, pPI3K and pAMPK and the mRNA expression levels of GLUT4 in adipocytes inducing insulin resistance increased in CL01-treated group compared with negative control. These results demonstrated that CL01 affected glucose metabolism and the protein and gene expression through insulin signaling pathway, and increased glucose uptake levels effectively. More than 90% of those who have suffered for type 2 diabetes are more likely to have from hyperinsulinemia, hypertension, obesity and etc. because of altered insulin signaling pathway. So, it is probably considered that intake of CL01 may treat type 2 diabetes by normalization of insulin signaling pathway, and it will provide useful evidences regarding a mechanism for cure of type 2 diabetes.

In vitro Anticancer Effect of Salt on HepG2 Human Hepatocellular Carcinoma Cells (소금의 HepG2 인체 간암세포에서의 in vitro 항암 효과)

  • Kim, Hee-Young;Ju, Jaehyun;Lee, Kyung Hee;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.137-142
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    • 2016
  • We investigated the anti-proliferative effects of solar salt and purified salt (PS) on HepG2 human hepatocellular cancer cells as well as their effects on mRNA and protein expression of apoptosis- and cell cycle-related genes, including Bcl-2, Bax, p53, and p21. Each salt sample suppressed cancer cell proliferation when treated at a concentration of 0.5% or 1%. Especially solar salt from T salt field (SS-T) and solar salt from Y salt field (SS-Y) significantly suppressed proliferation of cancer cells in comparison with PS. Treatment of HepG2 cells with salt samples at a concentration of 1% suppressed expression of Bcl-2 and promoted expression of Bax, p53, and p21 at the mRNA and protein levels in comparison with the control group. Inductively coupled plasma optical emission spectrometry (ICP-OES) showed that SS-T and SS-Y had higher concentrations of Ca, Mg, S, and K than PS, and SS-T contained higher concentrations of these minerals than SS-Y. It seems that Na and mineral contents in solar salt may contribute to regulation of the genes. Taken together, salt, especially mineral rich solar salt, inhibits cancer cell growth by regulating apoptosis and cell cycle-related genes.

Anti-inflammatory effects of ethanolic mulberry extract on the murine macrophage cell line, RAW 264.7 (RAW 264.7 큰포식세포에서 상백피 및 상지 에탄올 추출물의 항염증 활성 비교)

  • Kim, Yunyoung;Yang, Yoon Kyoung;Kim, Dongmin;Kim, Ji Yeon
    • Korean Journal of Food Science and Technology
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    • v.49 no.3
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    • pp.343-348
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    • 2017
  • The aim of this study was to compare the anti-inflammatory effects of ethanol extracts of root peel and spear of mulberry (RME and SME, respectively) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Both extracts significantly inhibited the production of reactive oxygen species (ROS), nitric oxide (NO), and interleukin-6 (IL-6). However, prostaglandin $E_2$ ($PGE_2$) levels decreased in LPS-stimulated RAW 264.7 cells treated with SME. Additionally, the extracts reduced inducible NO synthase (iNOS) expression and cyclooxygenase-2 (COX-2) in mRNA levels. Although ROS production was lower in the RME-treated cells than in the SME-treated cells, the levels of other inflammatory parameters, including IL-6 and $PGE_2$, and mRNA levels of iNOS and COX-2 reduced more in the SME-treated cells. These results indicate that SME showed higher anti-inflammatory activities than RME. Therefore, SME can be used as a functional food ingredient to enhance health.