• Title/Summary/Keyword: mRNA 발현

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Mechanism of Anti-inflammatory Effect of Peucedanum japonicum Thunb (방풍의 항염 효과 기전)

  • Noh, Sung-Il;Kim, Sang-Don;Park, Sung-Cheul;Seo, Byung-Yun;Yeom, Seung-Ryong;Kwon, Young-Dal;Shin, Byung-Cheul;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.15 no.2
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    • pp.31-43
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    • 2005
  • 목적 : 방풍은 임상적으로 관절염을 포함한 다양한 염증성 질환 치료제로 사용되어 왔다. 본 연구에서는 인간 비만세포를 이용하여 세포 독성에 영향을 주지 않는 농도에서 방풍의 항염 효과 및 그 기전을 검토했다. 방법 : 인간의 HMC-1세포를 IMDM에서 페니실린, 스트렙토마이신, 모노티오글리세린을 첨가하여 배양하고 방품추출액을 투여하였다. 그 다음 MTT, CLISA, RT-PCR, 세포내 칼슘측정, 핵단백분석을 이용하여 TNF-${\alpha}$, IL-6, IL-8 각각의 형성과 mRNA발현, 세포내 칼슘 수준, NF-${\kappa}B$ 발현에 대한 방풍추출액의 반응을 측정하고 통계처리 하였다. 결과 : 방풍은 PMA와 calcium ionophore A23187로 활성화된 비민세포에서 세포내 칼슘 수준과 NF-${\kappa}B$, TNF-${\alpha}$와 IL-6의 발현을 억제시켰고 RT-PCR을 이용한 mRNA 발현에서 TNF-${\alpha}$와 IL-6의 발현을 억제하였다. 결론 : 방풍은 비만세포내 칼슘 수준 및 NF-${\kappa}B$의 활성을 억제하고 염증성 세포 활성 물질인 TNF-${\alpha}$와 IL-6의 분비도 억제하여 항염효과를 나타냄을 암시하고 있다.

Effect on Inhibition of Matrix Metalloproteinase-1 in Human Dermal Fibroblasts by Production of Exopolysaccharide from Mycelial Culture of Grifola frondosa (잎새버섯이 생산하는 세포외 다당체의 사람 섬유아세포에서 Matrix Metalloproteinase-1 발현저해 효과)

  • Sim Gwan Sub;Bae Jun Tae;Lee Dong Hwan;Kim Jin Hwa;Lee Bum Chun;Choe Tae Boo;Pyo Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.2 s.51
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    • pp.161-167
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    • 2005
  • We investigated the effect on inhibition of matrix metalloproteinase (MMP) in human dermal fibroblast (HDF) by production of exopolysaccharide (GF-glucan) from mycelial culture of Grifola frondosa HB0071. The photoprotective potential of GF-glucan was tested in HDF exposed to ultraviolet-A (UVA) light. It was revealed that GF-glucan had an inhibitory effect on MMP-1 expression in UVA-irradiated HDF without any significant cytotoxicity. The treatment of UVA-irradiated HDF with GF-glucan resulted in a dose-dependent degrease in the expression level of MMP-1 protein and mRNA (by maximum $54.4\%$ at an $0.5\%$ GF-glucan). These results suggest that GF-glucan obtained from mycelial culture of G. frondosa HB0071 may contribute to inhibitory action in photoaging by reducing the MMP-1 related matrix degradation system.

Isolation and Characterization of a Wound or UV Induced cDNA Fragment from Pleurotus sajor-caju (상처 및 자외선 자극에 반응하는 여름느타리 cDNA 단편의 분리 및 그 발현 특성)

  • Park, Soo-Chul;Jung, Uk-Jin;Jeong, Mi-Jeong;Kim, Bum-Gi;Yoo, Young-Bok;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.314-320
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    • 1998
  • A 0.4 kb cDNA fragment was isolated from mRNA of UV or mechanical wound damaged Pleurotus sajor-caju by the differential display method. Expression of the gene corresponding to this cDNA fragment was highly induced by mechanical wound damage or UV treatment. This gene showed only basal level expression in mycelia, stipe, and cap under normal growth conditions. Sequencing analysis showed that this cDNA fragment contains partial open reading frame. Homology search using genbank database revealed that although this gene do not have homology with already reported wound induced genes, it has a significant sequence homology in defined region with the cdc2-related protein kinase gene which is known to be involved in negative regulation of meiotic maturation in Xenopus oocytes.

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THE EFFECT OF Fam83h KNOCKDOWN ON THE AMELOGENIN GENE EXPRESSION IN THE AMELOBLAST CELL LINE (Fam83h 발현 억제에 의한 조법랑세포 Amelogenin 발현 변화)

  • Lee, Sook-Kyung;Lee, Kyung-Eun;Kim, Jung-Wook
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.4
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    • pp.467-471
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    • 2010
  • Amelogenesis imperfecta, one of the dental genetic disease, is clinically and genetically complex disease. Amelogenesis imperfecta can be classified into three major categories according to clinical phenotype; hypoplastic, hypomaturation, and hypocalcification. Recently a novel gene, Fam83h, was identified to cause autosomal dominant hypocalcification amelogenesis imperfecta, however its functional role in the pathogenesis of enamel defect is not known yet. So this study was aimed to identify the knockdown effect of Fam83h gene on the amelogenin mRNA expression via shRNA transfection into immortalized ameloblast cell line. The result showed that the knockdown of Fam83h did not influence the amelogenin expression. Further study of the functional role of Fam83h gene should be performed to understand the complex nature of amelogenesis as well as molecular pathogenesis of amelogenesis imperfecta.

Effect of Korean Ginseng on the Expression of Transferrin Receptor in the Liver Cell Membrane of Rat (백서의 간세포막에서 Transferrin Receptor의 발현에 미치는 인삼의 영향)

  • Lim Jong-Ho;Hong Jang-Hee;Hur Gang-Min;Seok Jeong-Ho;Lee Jae-Heun
    • Journal of Ginseng Research
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    • v.23 no.2 s.54
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    • pp.105-114
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    • 1999
  • To investigate the effects of korean ginseng on the expression of transferrin receptor (TfR) in the liver cell membrane, we had carried out the experiments of $[^{3}H]thymidine$ uptake, $^{125}I-transferrin$ binding, and TfR mRNA expression in the liver after partial hepatectomy of normal and 3'-methyl-4-dimethylaminoa-zobenzene (3'-Me-DAB) treated rat with or without treatment of korean gingseng. $[^3H]thymidine$ uptake was not changed in the liver of 3'-Me-DAB or ginseng treated rat compared to that of control rat, but increased in that of partial hepatectomy of normal or 3'-Me-DAB treated rat. And this increased $[^{3}H]thymidine$ uptake was lowered slightly by the treatment of ginseng. Transferrin binding sites in the liver plasma membrane of ginseng treated rat with or without partial hepatectomy were similar, but increased in that of 3'-Me-DAB treated rat with or without partial hepatectomy compared to those of each control rat and these increased binding sites were reduced by ginseng treatment. Transferrin binding affinity (l/kd) was not changed by ginseng treatment, but tended to decrease in the liver of 3'-Me-DAB treated rat or in those after partial hepatectomy of all groups and reverse by ginseng treatment in 3'-Me-DAB treated rat. The expression of TfR mRNA was increased in the liver of 3'-Me-DAB treated rat with partial hepatectomy (peak at 24 hours), but lowered by ginseng treatment in this rat. From these results, it is suggested that korean ginseng has no effect on the increased expression of TfR with decreased affinity in the cell membrane of regenerated liver after partial hepatectomy of rat, but could inhibt that of 3'-Me-DAB treated rat through the regulation of DNA synthesis or TfR mRNA in partial.

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The Effect of Endotoxin on Gene Expression and Total Amount of Surfactant Protein A (내독소가 Surfactant Protein A의 유전자 발현과 총단백량에 미치는 영향에 관한 실험적 연구)

  • Moon, Doo-Seop;Sohn, Jang-Won;Yang, Seok-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.6
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    • pp.703-714
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    • 2000
  • Background : Surfactant protein A (SP-A) is important in the regulation of surfactant secretion, synthesis and recycling. SP-A has important roles in regulating surfactant metabolism as well as in determining surfactant's physical properties. Since systemic sepsis is one of the common causes of acute respiratory distress syndrome (ARDS) and abnormalities in surfactant function have been described in ARDS, the authors investigated the effects of endotoxemia on the accumulation of mRNA encoding SP-A and SP-A protein content. Methods : Adult rats were given various doses of intraperitoneal endotoxin from Salmonella enteritidis and sacrificed at different times. SP-A mRNA was measured by filter hybridization method. Lung SP-A protein content was determined by double sandwich ELISA assay using a polyclonal antiserum raised in rabbits against purified rat SP-A. Results : 1) The accumulation of SP-A mRNA in the endotoxin treated group 24 hours after 2mg/kg and 5mg/kg endotoxin treatments was significantly increased 50.9% and 27.3%, respectively, compared to the control group (P<0.001, P<0.025). 2) The accumulation of SP-A mRNA 24 hours in the 5mg/kg endotoxin treated group was significantly increased by 26.5% compared to the control group (P<0.01). 3) Total amount of lung SP-A was not altered at 24 hours by various doses of treatment. Total lung SP-A content 144 hours after endotoxin administration was significantly decreased by 51.4% compared to the control group (P<0.01). Conclusions : The specific regulation of SP-A by various time course in vivo is evident. The late decline in SP-A protein content was unexpected and suggests that SP-A may be differentially regulated during lung inflammation. The functional significance of these alterations remains to be clarified.

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Co-Culture Model Using THP-1 Cell and HUVEC on AGEs-Induced Expression of Cytokines and RAGE (THP-1 Cell과 HUVEC을 이용한 Co-Culture Model System에서 최종당화산물에 의한 Cytokines와 RAGE 발현)

  • Lee, Kwang-Won;Lee, Hyun-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.3
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    • pp.385-392
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    • 2011
  • Although monoculture methods have been remarkably useful due to their simplicity, they have serious limitation because of the different types of cells communication with each other in many physiological situations. We demonstrated levels of markers of endothelial dysfunction such as tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$) as well as stimulation of receptor of advanced glycation endproducts (AGEs) on monoand co-culture system such as only monocyte (THP-1) cultivation system, only endothelial cell (HUVEC) cultivation system, and co-cultivation system of THP-1 and HUVEC. The mRNA levels of TNF-$\alpha$ and IL-1$\beta$ on HUVEC increased by the co-culture with monocyte after 4 hr at 100 ${\mu}g/mL$ glyceraldehyde-AGE. The secreted protein contents into medium of TNF-$\alpha$ and IL-1$\beta$ increased after 8 hr approximately 2~2.5 times compared to mono-cultivation. In contrast, the mRNA level of receptor of AGE (RAGE) was relatively insensitive on the co-culture system. The mediators by which monocytes activate endothelial cell have not been fully elucidated. In this study we confirmed production of soluble cytokines such as TNF-$\alpha$ and IL-1$\beta$ by monocytes. Use of monocyte conditioned medium, which contains both cytokines, can activate endothelial cell.

Molecular Characterization of Metallothionein Gene of the Korean Bitterling Acheilognathus signifer (Cyprinidae) (묵납자루 (Acheilognathus signifer; Cyprinidae) metallothionein 유전자의 클로닝 및 특징 분석)

  • Lee, Sang-Yoon;Bang, In-Chul;Nam, Yoon-Kwon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.10-20
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    • 2011
  • Genetic determinant for metallothionein (MT), a cysteine-rich protein playing essential roles in metal detoxification and homeostasis, was characterized in the Korean bitterling (Acheilognathus signifer, Cyprinidae), an endemic fish species. The full-length A. signifer MT (AsMT) cDNA (551 bp) is composed of a single open-reading frame (ORF) to encode a polypeptide of 60 amino acids containing 20 cysteine residues whose positions are conserved in most cypriniform MTs. At the genomic level, the AsMT (2,593 bp spanning the 5'-flanking region to the 3'-untranslated region) represented a conserved tripartite (three exons interrupted by two introns) structure with AT-rich introns. The upstream regulatory region (-1,914 bp from the ATG initiation codon) of AsMT displayed various sites and motifs for transcription factors involved in the metal-mediated regulation and stress/immune responses. The AsMT transcript was ubiquitously detected in various organs with variable expression levels, where the ovary and intestine showed the highest expression, while the heart and skeletal muscle represented the lowest level. During an exposure to copper (immersion in $0.5\;{\mu}M$ Cu for 48 h), the levels of AsMT transcripts were significantly elevated in the liver (more than 3.5-fold), moderately in the gill, kidney, and spleen (ranging from 1.5- to 2.5-fold), and barely in the brain and intestine. Results of this study could form a useful basis to explore the metal-related stress physiology of this endangered fish species.

A study of the lipoprotein lipase inhibitory mechanism of Poncirus trifoliata water extracts (탱자 (Poncirus trifoliata)의 lipoprotein lipase 억제메커니즘)

  • Lee, Sung Mee;Kang, Yun Hwan;Kim, Kyoung Kon;Kim, Tae Woo;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.48 no.1
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    • pp.9-18
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    • 2015
  • Purpose: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. Methods: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein ($C/EBP{\beta}$). Results: The total polyphenol and flavonoid content of PF-W was $52.15{\pm}4.02$ and $6.56{\pm}0.47mg/g$, respectively. PF-W treatment decreased LPL content in media to $58{\pm}5%$ of that in control adipocyte media, and increased LPL content to $117{\pm}3.5%$ of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of $C/EBP{\beta}$, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. Conclusion: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.

Characterization of a cDNA Encoding Transmembrane Protein 258 from a Two-spotted Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus)의 GbTmem258 cDNA 클로닝과 발현분석)

  • Kisang Kwon;Honggeun Kim;Hyewon Park;O-Yu Kwon
    • Journal of Life Science
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    • v.33 no.10
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    • pp.828-834
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    • 2023
  • The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.