• Title/Summary/Keyword: mRNA 발현

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The effects of antioxidant and anti-aging treatment of UVB-irradiated human HaCaT keratinocytes with ethanol extracts of colored rice varieties (유색미 에탄올 추출물의 산화방지 효과와 UVB에 유도된 인간유래 피부각질형성세포에서의 항노화 효과)

  • Choi, Eun-Young;Lee, Jin-Tae
    • Korean Journal of Food Science and Technology
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    • v.50 no.6
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    • pp.653-659
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    • 2018
  • In this study, we investigated the antioxidant activities and anti-aging efficacy in terms of suppression of matrix metalloproteinases (MMPs) in UVB-irradiated HaCaT cells by adding the ethanol extracts of Josaengheogchal (JE) and Shintoheug rice (SRE). In the electron-donating ability and ABTS radical-scavenging assays, we observed that both JE and SRE had scavenging activities and in a collagenase inhibition assay, both extracts showed inhibition effects of over 73% at $1,000{\mu}g/mL$ concentration. The expression of MMP-1 and -3, when the extracts were treated with UVB $50mJ/cm^2$, irradiated human HaCaT keratinocytes, was analyzed by western blotting and reverse-transcription polymerase chain reaction (RT-PCR). The results showed that MMP-1 and -3 proteins and mRNAs were downregulated in a concentration-dependent manner in response to both extracts. Therefore, we expect that these compounds have a potential for the use as functional ingredients with anti-aging effects in the cosmetic and food industries.

Control Mechanisms of Ovulation by Pituitary Adenylate Cyclase-Activating Polypeptide (Pituitary Adenylate Cyclase-Activating Polypeptide에 의한 배란 조절에 관한 연구)

  • Lee, Yu-Il;Kim, Hyoung-Choon;Kim, Mi-Young;Chun, Sang-Young
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.101-111
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    • 2005
  • 배 경: Pituitary adenylate cyclase-activating polypeptide (PACAP)은 양의 시상하부에서 추출된 신경펩타이드 호르몬으로 난소에도 존재하여 배양된 과립막 세포에서 스테로이드합성과 cyclic AMP 형성을 촉진함이 보고되었다. 목 적: 흰쥐 난소를 실험 모델로 사용하여 배란시 황체화호르몬 (luteinizing hormone; LH)에 의해 유도된 PACAP과 PACAP 수용체의 유전자 발현양상과 신호 전달경로를 규명하고자 하였다. 재료 및 방법: 미성숙 흰쥐의 배란전 난포를 체외 배양하면서 LH로 처리하고 PACAP 및 PACAP수용체의 유전자 발현을 보기 위해서는 Northern blot 분석과 in situ hybridization (ISH)을, 그리고 단백질 수준의 PACAP 검색을 위해서는 enzyme linked immunosorbent assay (ELISA) 분석을 이용하였다. 결 과: LH 처리 후 Northern blot상의 PACAP 유전자 발현은 6~9시간에 일시적으로 최고치에 도달하였으며 ISH로 보아 과립막 세포에서 발현됨을 알 수 있었다. ELISA 분석 상 PACAP 단백질도 LH처리 후 6~12시간에 최고치를 나타내었으며, PACAP 수용체 mRNA 역시 3~9시간에 최고치로 과립막 세포에서 발현되었다. Adenylate cyclase (AC) 억제제인 MDL12330A 처리시 LH로 발현된 PACAP mRNA가 감소되며, AC의 활성제인 forskolin 처리에는 LH시와 유사한 PACAP mRNA의 발현양상을 나타내었다. 그러나 protein kinase C (PKC)의 억제제인 chelerythrine과 2-0-tetradecanolphorbol-13-acetate (TPA) 처리로는 PACAP 의 유전자 발현에 영향을 주지 못하였다. 5-lipoxygenase의 억제제인 MK886이나 nordihydroguaiaretic acid (NDGA)로 처리한 결과 LH로 유도된 PACAP 유전자의 발현이 감소되었으나, cyclooxygenase의 억제제인 indomethacin은 별로 영향을 주지 못하였다. MEK와 p38의 억제제인 PD98059와 SB203580도 LH로 촉진 된 PACAP의 유전자 발현을 농도 의존적으로 억제하였다. 결 론 : 배란전 난포에서 PACAP과 PACAP 수용체의 유전자 발현은 모두 LH의 폭발적 분비에 의해 유도되어 일시적으로 과립막 세포에서 나타나 배란을 위한 국소적인 조절 작용을 할 것으로 추정되며, LH로 촉진된 PACAP 유전자 발현을 위한 신호전달은 cAMP-PKA, lipoxygenase 및 MAP kinase 경로를 통하는 것으로 사료된다.

Expression of Toll-like Receptor-2 on the Peripheral Blood Monocytes in Kawasaki Disease Patients (가와사끼병 환자에서 분리한 CD14양성 세포에서 Toll-like Receptor-2의 발현)

  • Hwang, Dae Hwan;Han, Jung Woo;Choi, Kyung Min;Shin, Kyung Mi;Kim, Dong Soo
    • Clinical and Experimental Pediatrics
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    • v.48 no.3
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    • pp.315-320
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    • 2005
  • Objective : Toll like receptor(TLR) is known to be involved in innate immunity. Many microbial antigens stimulate TLR, and as a result of intracellular signal transduction, they activate nuclear factor-kB which produces diverse inflammtory cytokines. Until now, many research topics in Kawasaki disease focused on cytokine increasement. In this study, we aim to reveal TLR increasement which might be associated with initiation of inflammatory response. Methods : We obtained the peripheral blood of ten patients who were diagnosed with Kawasaki disease in Yonsei University College of Medicine from March 2003 to August 2003, as well as those of a febrile control group and the same number of a normal control group. Flow cytometry was done in all samples for quantification of TLR-2 expression in CD14 positive monocyte. And we also extracted total RNA of periphral monocyte and quantificated expression of TLR-2 mRNA by RT-PCR. Results : The expression of TLR-2 in Kawasaki disease increased significantly compared with the normal control group but not when compared with the febrile control group. And the expression decreased slightly in the subacute phase of Kawasaki disease compared with the acute phase, but this was statistically insignificant. mRNA expression of TLR-2 in peripheral blood monocyte also increased in the acute phase of Kawasaki disease. Conclusion : Expression of TLR-2 in Kawasaki disease increased when compared with the normal control group, which means that innate immunity is associated with the pathogenesis of Kawasaki disease.

Effect of [6]-Gingerol on Bcl-2 and Bax Expression in MDA-MB-231 Human Breast Cancer Cell Line ([6]-Gingerol이 인체 유방암세포 MDA-MB-231에서 Bcl-2와 Bax 발현에 미치는 영향)

  • Seo, Eun-Young;Kim, Woo-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.6
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    • pp.671-676
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    • 2006
  • We investigated the effect of gingerol (Zingiber officinale Roscoe, Zingiberaceae) on Bcl-2 and Bax expression in MDA-MB-231 human breast cell lines. The oleoresin from rhizomes of ginger contains [6]-gingerol (1-[4'-hydroxy-3'-methoxyphenyl]-5-hydroxy-3-decanone). We previously reported that [6]-gingerol inhibits cell proliferation in MDA-MB-231 human breast cancer cell lines. In this study, we examined protein and mRNA expression associated with cell apoptosis in MDA-MB-231 human breast cancer cell lines. We cultured MDA-MB-231 cells in presence of various concentrations 0, 2.5, 5 and $10\;{\mu}M$ of [6]-gingerol. Bcl-2 protein and its mRNA levels were decreased dose-dependently in cells treated with [6]-gingerol, but Bax protein and its mRNA levels were unchanged by [6]-gingerol treatment. Bcl-2/Bax ratio was decreased in a dose dependent manner treated with [6]-gingerol. Caspase-3 activity was significantly increased dose-dependently in cell treated with [6]-gingerol (p<0.05). In conclusion, we have shown that [6]-gingerol induces apoptosis in MDA-MB-231 human breast cancer cell lines.

A Study on the Development of an Early Embryonic Gene of the Silkworm, Bombyx mori (누에 배형성기 초기 발현 유전자 개발 연구)

  • Choi, Kwang-Ho;Goo, Tae-Won;Kim, Seong-Ryul;Park, Seung-Won;Kim, Sung-Wan;Kang, Seok-Woo
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.122-125
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    • 2012
  • This study was aimed for a development of a useful gene promoter which has a transcript expressional specificity in the early embryonic period of the silkworm, Bombyx mori. To select a useful gene expressed in the early embryonic stage, we constructed and analyzed a PCR-base subtraction cDNA library. In subtractive hybridization analysis, we confirmed four clones as differently expressed genes(BmNanos-like, BmNanos-P, BmNanos-O, BmVasa mRNAs). Northern hybridization and real time PCR results reveled that the BmNanos-like gene promoter is suitable for the silkworm transgenic vector system. Further defined studies on molecular functions and biological roles of their promoters will give us well-fined information and its application.

Expression of Leptin and Its Receptor in Rat Ovary (흰쥐 난소내 Leptin 및 Leptin 수용체의 발현)

  • 김명신;양현원;권혁찬;황경주;윤현숙;박금자;김세광;윤용달
    • Development and Reproduction
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    • v.2 no.2
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    • pp.173-178
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    • 1998
  • Leptin, the product of the obese gene, is produced by adipose tissue and is known to be a hormone concerned with regulation of appetite and metabolism. Recent reports have shown that leptin is associated not only with obesity but also with female reproduction, but it has not yet been ascertained whether leptin acts directly on the ovaries or indirectly via the hypothalamus or pituitary pathway. The object of this study is to determine the expression of leptin and its receptor in the ovaries of 3 and 8 weeks old rats by immunohistochemistry and RT-PCR. In the ovaries of 3 and 8 weeks old rats, leptin was stained in the theca cells and portions of granulosa cells of atretic follicles, whereas leptin receptors was stained in interstitial cells and ova of preantral follicles. The RT-PCR results showed that leptin receptor mRNA was expressed in the ovaries of both immature and adult rats, while leptin mRNA was not. In conclusion, leptin mRNA was not expressed in the ovaries, however, leptin was detected by immunohistochemistry. Compared to leptin itself, leptin receptors in the ovaries were ascertained by both RT-PCR and immunohistochemistry. These results suggest that leptin is related to the regulation of the physiological functions of the ovaries.

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Alteration of Insulin-like Growth Factor(IGF)-I and IGF-Binding Proteins in Renal Development and Regeneration (신장발육 및 재생에 따른 insulin-like growth factor(IGF)-I 및 IGF-binding protein의 변화)

  • Park Sung-Kwang;Koh Gou-Young;Lee Dae-Yeol
    • Childhood Kidney Diseases
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    • v.3 no.2
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    • pp.109-116
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    • 1999
  • Purpose: Insulin-like growth factor(IGF)-I and -II are peptide growth factor whose activity is modulated by interaction with the family of six IGF-binding proteins(IGFBPs). IGF-I is detected in rat kidney and has metabolic and growth effects. This study was designed to examine temporal expression of IGFBPs in kidney during renal development and postischemic regeneration in rat. Method: The expression of IGFBPs in kidney during renal development from 15th day of gestation to adult life by using Northern blot analysis. We also examined the renal IGF-IGFBP axis in uremic rat by using Northern blot and immunohistochemistry. Results: The mRNA of IGFBP-1 and -3 were not or barely detected in fetal stages. However, the mRNA level of IGFBP-1 and -3 were increased gradually from day 7 after birth to adult. In contrast, the mRNA of IGFBP-2 and -5 were highly expressed in fetal stages and maintained almost same levels until day 7 (IGFBP-2) or day 30 (IGFBP-5) after birth, then their levels decreased markedly. The mRNA of IGFBP-4 were expressed moderately in fetal kidney and increased gradually after birth. Interestingly, the mRNA of IGFBP-1 and-4 were induced up to 3-5 fold during maximum regeneration period and were recovered to normal levels after acute ischemic injury. In contrast, the mRNA level of IGFBP-3 and-IGFBPrP-1 were decreased slightly at 1 day after ischemic injury, then recovered to normal level during maximum regeneration period. Conclusion: There were differential expressions of IGFBPs in kidney that can modulate IGF action on developing, differentiating, maintaining, and regenerating renal structure and function.

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Characterization of Mud Loach (Misgurnus mizolepis) Apolipoprotein A-I: cDNA Cloning, Molecular Phylogeny and Expression Analysis (미꾸라지(Misgurnus mizolepis) Apolipoprotein A-I cDNA의 구조, 분자계통 및 발현 특징 분석)

  • Lee, Youn-Ho;Noh, Jae-Koo;Kim, Keun-Yong;Cho, Young-Sun;Nam, Yoon-Kwon;Kim, Dong-Soo
    • Journal of Aquaculture
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    • v.20 no.1
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    • pp.65-72
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    • 2007
  • Full length complementary DNA encoding apolipoprotein A-I (apoA-I) was isolated and characterized in mud loach (Misgurnus mizolepis). Mud loach apoA-I cDNA encoding 24 bp of 5'-untranslated region (UTR), 762 bp of single open reading frame (ORF) consists of 254 amino acids and 293 bp of 3'-UTR excluding stop codon and poly (A+) tail. Two overlapping polyadenylation signals (AATAAAATAAA) was found 9 bp prior to the poly (A+) tail. Mud loach apoA-I represented considerable homology to those from other teleost species at amino acid level with conserving common features of vertebrate apoA-I. Molecular phylogenetic analysis inferred the phylogenetic hypothesis that was generally in accordance with the previous taxonomic relationship. Apolipoprotein A-I mRNA was detected in various tissues, but the mRNA levels were quite varied depending on tissues based on semi-quantitative RT-PCR. Liver and brain showed the significantly higher levels of apoA-I transcripts than other tissues. mRNA expression of apoA-I was quite low in very early stage of embryonic development, however dramatically enhanced from 8 hours post fertilization. This increased mRNA level was retained consistently up to 14 days post hatching.

The Effect of Interleukin-10 on KC Gene Expression in Mouse Peritoneal Macrophages (케모카인 KC 유전자 발현에 대한 Interleukin-10의 억제작용)

  • Kim, Hee-Sun
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.47-54
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    • 1998
  • Interleukin-10(IL-10) inhibits production of a wide range of cytokines in various cell types and transcriptionally inhibits lipopolysaccharide (LPS)-induced expression of proinflammatory mediators. Cytokine expression by macrophages is an important aspect to ochestrate inflammatory responses. As an approach to identify mechanistic targets of IL-10, it was examined the time course for expression of KC(murine homologue of Gro) gene in murine peritoneal macrophages stimulated with LPS with or without IL-10. The effect of IL-10 on LPS induced KC mRNA expression was delayed and only seen after 1 hour treatment. Pretreatment with IL-10 did not eliminate the delayed inhibitory response nor increase the magnitude of suppression. These effects did not depend upon time of IL-10 treatment but the time of LPS treatment. LPS-induced KC mRNA expression by inhibitory action of IL-10 was not controlled at the level of transcription. The result indicates that IL-10 acts late in the process of KC gene expression and that the prominant site of action may be mRNA stability or translation.

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Changes of Gill Structure and Identification of Genes by Muddy Water Exposure in Cyprinus carpio (잉어(Cyprinus carpio)에서 탁수 노출에 의한 아가미 미세구조 변화와 유전자 확인)

  • Shin, Myung-Ja;Lee, Jong-Eun;Seo, Eul-Won
    • Korean Journal of Ecology and Environment
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    • v.44 no.1
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    • pp.95-101
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    • 2011
  • Present study aimed to investigate morphological change and gene expression in the gill of Cyprinus carpio after exposure against muddy water caused by riverbed disturbance with various rearing condition. The gill of C. carpio showed abnormal shapes in its secondary lamellae and a rough surface with impure debris after exposed to muddy water for 80 days. In addition, the gills showed the edema, the exfoliation of epithelial cell, and the fusion of the secondary lamellae. Using 20 ACPs, 24 differentially expressed genes (DEGs) exposed to muddy water for 80 days were identified. 17 genes among them were up-regulated, while 7 genes were down-regulated in preterm deliveries. A BLAST searches revealed that 3 genes were revealed known genes as calcium transporter 1 (TRPV6) mRNA, macha mRNA for putative puroindoline b protein, and Efnb3 protein-like. Therefore, it is considered that Efnb3 gene from gill would be a useful indicator for neurobehavioral changes in fish influenced by muddy water.