• Title/Summary/Keyword: mRNA 발현

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Expression Analysis of Matrix Metalloproteinases and Tissue Inhibitor of Matrix Metalloproteinases from In Vitro Maturation Oocytes Complexes in Porcine (돼지 체외성숙난자에서 MMPs와 TIMPs의 발현 분석)

  • Kim, Sang-Hwan;Kang, Hyun-Ah;Kim, Dae-Seung;Lee, Myeong-Seop;Seo, Kang-Suk;Min, Kwan-Sik;Yoon, Jong-Taek
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.55-62
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    • 2010
  • Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, DC and CDCs. Activity of MMP-2 in the DC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the CDCs 36 hr. Expression of TIMP-3 protein in the CDCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

Effects of High Fat Diet on Serum Leptin and Insulin Level and Brown Adipose Tissue UCP 1 Expression in Rats (흰쥐에서 고지방식이가 혈중 렙틴 및 인슐린과 갈색지방조직의 UCP 1 발현에 미치는 영향)

  • 홍경희;강순아;김소혜;조여원
    • Journal of Nutrition and Health
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    • v.34 no.8
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    • pp.865-871
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    • 2001
  • The adipose tissue hormone leptin has been proposed to be involved in the regulation of flood intake and energy expenditure via thermogenesis by uncoupling protein(UCP) in brown adipose tissue(BAT). The objective of the study was to examine the effects of high fat diet on the serum leptin levels, BAT UCPl expression and the body fat mass in rats after weaning. During experimental period of 12 weeks, 4 male Sprague-Dawley rats were killed for the baseline experiment at 4 weeks of age while the remaining rats were fed the two different diets: the control diet AIN-76A(n = 20), high fat(beef tallow) diet(n = 20) ad libitum, which provided 11.7% or 40% of calories as fat, respectively. At 16 weeks of age, the increase in the food efficiency ratio(FER) was related to fat mass in rats on high fat diet. Serum leptin level was increased by age and dietary high fat. There was no difference in serum insulin level between groups until 10 weeks of age, but rats fed high fat diet for 12 weeks showed hyperinsulinemia. The amount of body fat pads was increased significantly in high fat group compared to normal diet group. Visceral fat mass affected acutely by high fat diet, as a result, it was higher in rats fed high fat diet for 2 weeks than normal diet. At 16 weeks of age, BAT and visceral fat mass were significantly high in high fat group. Also, the serum leptin levels reflected the amount of body fat mass. BAT UCPI mRNA expression increased with age and dietary high fat. This study demonstrates that dietary high fat increased serum leptin levels, BAT UCPI expression and body fat mass. Futhermore, in rats fed high fat diets, the increases in leptin and UCPI expression counteracts only in part the excess adiposity and obesity.

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Expression of Tryptophan Hydroxylase in the Hypothalamus and Hippocampus of Fasting and Anorexia Mutant Mice (절식시킨 생쥐와 식욕부진 돌연변이 생쥐의 시상하부와 해마에서의 Tryptophan Hydroxylase의 발현)

  • 김미자;김영옥;정주호
    • Journal of Nutrition and Health
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    • v.33 no.1
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    • pp.5-12
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    • 2000
  • The control of food intake is a complex phenomenon caused by interactions between central and peripheral control mechanisms. The hypothalamic and brain stem regions have been identified as centers for food intake and energy expenditure in animals and humans. Of these, the ventromedial and lateral hypothalamic areas are involved in the control of food intake. Also, large amounts of neurotransmitters known to be involved in feeding are present in the hippocampus. Paricularly, tryptophan hydroxylase(TPH), known as a factor in the control of food intake, is present in high levels in the paraventricular nucleus of the hypothalamus and the hippocampus. In this study, TPH expression levels in the hypothalamic and hippocampal regions of fasting, anorexia mutant, and control mice were compared using RT-PCR and immunohistochemical methods. Differences in body weight among the fasting, anorexia mutant, and control groups wire observed. No statistical significance was noted in the number of TPH-immunoactivity in the hypothalamic nuclei, but relatively higher populations of such fibers were observed in the fasting group : the control group yielded samples with an overall value of 170.3${\pm}$3.5 in terms of immunoreactivity-induced optical density, whereas the fasting group yielded a value of 168.3${\pm}$2.6, and the anorexia mutant group 171.3${\pm}$0.8(lower values represent higher immunoreactivity), In fasting mice, stained neuronal bodies were observed in the CA3 and dentate gyrus regions of the hippocampus, which was different from the hippocampal regions of the control and anorexia mutant mice. The RT-PCR procedures were performed using whole brains, precluding any statistically noticeable findings in relation to specific regions, although the fasting and anorexia mutant groups showed 123.3% and 102.9%, respectively, of the TPH mRNA level in the control. The overall results present evidences of the role of TPH in the decrease in food intake during fasting caused by exogenic factors and in genetically acquired anorexia. (Korean J Nutrition 33(1) : 5-12, 2000)

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Prediction and Annotation of ABC Transporter Genes from Magnaporthe oryzae Genome Sequence (벼도열병균 게놈서열로부터 ABC transporter 유전자군의 예측 및 특성 분석)

  • Kim, Yong-Nam;Kim, Jin-Soo;Kim, Su-Young;Kim, Jeong-Hwan;Lee, Jong-Hwan;Choi, Woo-Bong
    • Journal of Life Science
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    • v.20 no.2
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    • pp.176-182
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    • 2010
  • Magnaporthe oryzae is destructive plant-pathogenic fungus and causes rice blast. The pathogen uses several mechanisms to circumvent the inhibitory actions of fungicides. ATP-binding cassette (ABC) transporters are known to provide protection against toxic compounds in the environment. PC facilitated bioinformatic analysis, particularly with respect to accessing and extracting database information and domain identification. We predicted ABC transporter genes from the M. oryzae genome sequence with computation and bioinformatics tools. A total of thirty three genes were predicted to encode ABC transporters. Three of thirty three putative genes corresponded to three known ABC transporter genes (ABC1, ABC2 and ABC3). Copy numbers of the ABC transporter genes were proven by Southern blot analysis, which revealed that twenty genes tested exist as a single copy. We amplified the DNA complementary to RNA corresponding to eleven of these by reverse transcriptase polymerase chain reaction.

Cloning of Growth Hormone Complementary DNA from Red-Spotted Grouper (Epinephelus akaara) and Its Expression in E. coli (붉바리(Epinephelus akaara)의 성장호르몬 cDNA의 Cloning과 E. coli에서의 발현)

  • 강거영;송춘복;이제희
    • Journal of Aquaculture
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    • v.16 no.2
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    • pp.110-117
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    • 2003
  • We have cloned and sequenced the cDNA encoding growth hormone (GH) from pituitary poly(A)$^{+}$ RNA of red-spotted grouper (Epinephelus akaara). The cDNA of red-spotted grouper GH is 883 base pairs (bp) consisting of 21 bp of 5'untranslated region (UTR), 615 Up of an open reading frame (ORF) and 247 Up of 3'UTR. The polyadenylation signal, AATAAA, was 20 bp upsteam of polyadenylation site. Based on the nucleotide sequences, the deduced putative polypeptide contains 204 amino acids (aa), representing 17 aa of a signal and 187 aa of a mature polypeptide. The putative GH cDNA encodes a polypeptide with four cysteine residues and only one N-gly- cosylation site. Comparative sequence alignment shows that red-spotted grouper GH exhibits high similarity with its corresponding other Perciformes species GH cDNAs.

Stromal cell-derived factor-1 (SDF-1) expression in the oral squamous cell carcinoma (구강편평상피암종에서 stromal cell-derived factor-1의 발현)

  • Kim, Kyung-Wook;Han, Se-Jin;Roh, Kyu-Seob
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.1
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    • pp.1-6
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    • 2010
  • Purpose: Chemokines are structurally related, small polypeptide signaling molecules that bind to and activate a family of transmembrane G protein-coupled receptors, the chemokine receptors. Recently, interaction between the chemokine receptor CXCR4 and its ligand, stromal cell-derived factor 1 (SDF-1 or CXCL12), has been found to play an important role in tumorigenicity, proliferation, metastasis and angiogenesis in many cancers such as lung cancer, breast cancer, melanoma, glioblastoma, pancreatic cancer and cholangiocarcinoma. Hence, the goal of this study is to identify the correlation of clinicopathological factors and the up-regulation of SDF-1 expression in oral squamous cell carcinoma. Material and methods: We studied the immunohistochemical staining of SDF-1, quantitative RT-PCR (qRT-PCR) of SDF-1 gene in 20 specimens of 20 patients with oral squamous cell carcinoma. Results: 1. In the immunohistochemical study of poor differentiated and invasive oral squamous cell carcinoma, the high level staining of SDF-1 was observed. And the correlation between immunohistochemical SDF-1 expression and tumor nodes metastases (TNM) classification of specimens was significant.($x^2$ test, P < 0.05) 2. In the SDF-1 gene qRT-PCR analysis, SDF-1 expression was more in tumor tissue than in carcinoma in situ tissue. Paired-samples analysis determined the difference of SDF-1 mRNA expression level between the cancer tissue and the carcinoma in situ tissue.(Student's t-test, P < 0.05) Conclusion: These findings suggest that up-regulation of the SDF-1 may play a role in progression and invasion of oral squamous cell carcinoma.

High-fat Intake is Associated with Alteration of Peripheral Circadian Clock Gene Expression (고지방식이에 의한 말초 생체시계 유전자 발현 변화)

  • Park, Hyun-Ki;Park, Jae-Yeo;Lee, Hyangkyu
    • Journal of Korean Biological Nursing Science
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    • v.18 no.4
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    • pp.305-317
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    • 2016
  • Purpose: Recent studies demonstrated disruption of the circadian clock gene is associated with the development of obesity and metabolic syndrome. Obesity is often caused by the high calorie intake, In addition, the chronic stress tends to contribute to the increased risk for obesity. To evaluate the molecular mechanisms, we examined the expression of circadian clock genes in high fat diet-induced mice models with the chronic stress. Methods: C57BL/6J mice were fed with a 45% or 60% high fat diet for 8 weeks. Daily immobilization stress was applied to mice fed with a 45% high fat for 16 weeks. We compared body weight, food consumption, hormone levels and metabolic variables in blood. mRNA expression levels of metabolic and circadian clock genes in both fat and liver were determined by quantitative RT-PCR. Results: The higher fat content induced more severe hyperglycemia, hyperlipidemia and hyperinsulinemia, and these results correlated with their relevant gene expressions in fat and liver tissues. Chronic stress had only minimal effects on metabolic variables, but it altered the expression patterns of metabolic and circadian clock genes. Conclusion: These results suggest that the fat metabolism regulates the function of the circadian clock genes in peripheral tissues, and stress hormones may contribute to its regulation.

Effects of Lithospermum erythrorhizon on the cytokine gene expression in human keratinocytes (자초(紫草)가 HaCaT 세포의 사이토카인 유전자 발현에 미치는 영향)

  • Kang, Sang-Hoon;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.50-62
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    • 2013
  • Objective : Propionibacterium acnes (P. acnes) is a major pathogenic bacteria for acne vulgaris. This study was performed to evaluate the effects of Lithospermum erythrorhizon extracts on the inflammatory cytokines gene expression by P. acnes in human keratinocytes, HaCaT cell line. Methods : Anti-bacterial activity and cytotoxicity of LE extracts was analyzed by agar plate culture and XTT assay. The cytokines gene expressions were assessed by real time RT-PCR for IL-8, MCP-1 and TNF-${\alpha}$. During the cell culture and treatments, amounts of secreted TNF-${\alpha}$ were measured by ELISA. Translocation of transcription factor NF-${\kappa}B$ from cytoplasm into nucleus was observed by immunocytochemistry and confocal microscopy. Results : There were no anti-bacterial effects and cytotoxicity as high as $1,000{\mu}g/ml$ of LE extracts in XTT assay. Transcription levels of inflammatory cytokines, IL-8, MCP-1 and TNF-${\alpha}$ were increased by P. acnes in HaCaT. LE extracts decreased the upregulated gene transcription levels. However, amounts of secreted TNF-${\alpha}$ were similar in HaCaT cells with P. acnes and LE extracts. Translocation of NF-${\kappa}B$ into nucleus by P. acnes was significantly inhibited by LE extracts. Conclusions : From the results of this study, LE extracts have anti-inflammatory effects on HaCaT cells by P. acnes that decreased the mRNA expressions of IL-8, MCP-1 and TNF-${\alpha}$. This anti-inflammatory effects of LE extracts could provide the potential of therapeutic substance for acne vulgaris.

Antioxidant and anti-inflammatory effects of the ethyl acetate fraction of the Agastache rugosa extract (배초향 에틸아세테이트 분획의 산화방지 및 항염증 활성)

  • Kim, Bomin;Han, Yeong Eun;Lee, Hwa Jin
    • Korean Journal of Food Science and Technology
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    • v.49 no.3
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    • pp.331-337
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    • 2017
  • To evaluate the antioxidant activity of hexane, ethyl acetate, and butanol fractions obtained from Agastache rugosa extract, we measured the total polyphenol levels, DPPH radical scavenging activity, and reducing power. The ethyl acetate fraction of A. rugosa (AREA) displayed high phenolic levels, potent DPPH radical scavenging effect, and powerful reducing power. In addition, we examined the ability of AREA to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated BV-2 microglia. AREA suppressed NO production and inducible nitric oxide synthase (iNOS) expression and downregulated interleukin-6 (IL-6) mRNA level in LPS-stimulated BV-2 microglia. Furthermore, we detected rosmarinic acid in AREA by HPLC, which suggested that rosmarinic acid could be one of the bioactive materials responsible for the antioxidant and anti-inflammatory activities of AREA. These results suggested that AREA may be a good source of functional foods with antioxidant and anti-inflammatory activities.

The Effect of Heparin on Gene Expression of Collagen, MMP, TGF-β Isoforms Using Cultured Fibroblast from Transverse Carpal Ligament of Carpal Tunnel Syndrome (헤파린이 수근관 증후 군 환자의 횡수근 인대에서 추출한 섬유아세포의 콜라겐, MMP 및 TGF-β 동위형 발현에 미치는 영향)

  • Jeong, Seong Ho;Na, Min Hwa;Lee, Jae Sun;Song, Sang Hun;Yoon, Eul Sik;Dhong, Eun Sang;Kim, Han Joong
    • Archives of Plastic Surgery
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    • v.33 no.4
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    • pp.427-432
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    • 2006
  • Purpose: Noninflammatory synovial fibrosis has been noted for main causal factor of carpal tunnel syndrome (CTS). Recently, there are some reports that heparin have not only anti-coagulative effect but also anti-inflammatory and anti-fibrotic potential and have an effect on interstitial pulmonary fiborosis. Authors examined whether heparin affects pathogenesis of CTS. Methods: First, heparin was administered to fibroblast that was cultured from patient's transverse carpal ligament. Secondly, we evaluated the expression from genes of type I, III collagen, TGF ${\beta}$ isoforms and MMP. Fibroblasts were isolated and cultured from transverse carpal ligaments of 5 patients with CTS. Heparin (0, 1, 10,$100{\mu}g/ml$) was administered to cultured fibroblast and reverse transcription PCR for mRNA expression of type I, III collagen, TGF-${\beta}$ isoforms and MMP was done. Results: Heparin suppressed gene expression of type I, III collagen and TGF-${\beta}1$, ${\beta}3$ but promoted gene expression of TGF-${\beta}2$ and MMP-2. Conclusion: Heparin directly suppress gene expression of type I, III collagen. But, It is undetermined that heparin can present it's effect mediated by TGF ${\beta}$ isoforms or MMP.