• 제목/요약/키워드: mRNA 발현

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

  • 김상환;강현아;김대승;이명섭;서강석;민관식;윤종택
    • Reproductive and Developmental Biology
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    • 제34권1호
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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.

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

  • 홍경희;강순아;김소혜;조여원
    • Journal of Nutrition and Health
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    • 제34권8호
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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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절식시킨 생쥐와 식욕부진 돌연변이 생쥐의 시상하부와 해마에서의 Tryptophan Hydroxylase의 발현 (Expression of Tryptophan Hydroxylase in the Hypothalamus and Hippocampus of Fasting and Anorexia Mutant Mice)

  • 김미자;김영옥;정주호
    • Journal of Nutrition and Health
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    • 제33권1호
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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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벼도열병균 게놈서열로부터 ABC transporter 유전자군의 예측 및 특성 분석 (Prediction and Annotation of ABC Transporter Genes from Magnaporthe oryzae Genome Sequence)

  • 김용남;김진수;김수영;김정환;이종환;최우봉
    • 생명과학회지
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    • 제20권2호
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    • pp.176-182
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    • 2010
  • 벼의 생산에 있어 가장 큰 문제 요인 중 하나인 벼도열병의 발생 원인균인 벼도열병균은 다양한 기작에 의해 방제 약제에 대한 내성을 가지는 것으로 알려져 있다. 막 운반단백질인 ABC transporter의 경우 환경으로부터의 다양한 독성 물질들을 배출하는 것으로 알려져 있다. 이미 알려진 벼도열병균의 게놈 서열로부터 생물정보학적 분석을 통하여 ABC transporter 단백질의 도메인 특성을 보이는 33개의 유전자군 서열을 예측하였다. 이중 3개의 경우는 이미 알려진 유전자로 판명되었다. Southern Hybridization 분석에 적용한 20개의 유전자들이 모두 게놈상에 단일 copy로 존재함을 확인하였다. 새로 예측된 30개의 유전자중 11개는 RT-PCR을 통하여 전사단계에서의 유전자 발현이 확인되었다.

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

  • 강거영;송춘복;이제희
    • 한국양식학회지
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    • 제16권2호
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    • pp.110-117
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    • 2003
  • 붉바리(E. akaara)의 뇌하수체에서 추출한 mRNA로부터 RACE 방법으로 cDNA를 cloning하고 염기서열을 분석하였다. 붉바리의 성장호르몬 cDNA는 5' UTR, open reading frame, 3' UTR이 각각 21 bp, 615 bp, 247 bp인 전체 883 bp로 구성되어있다. Adenylation signal로 작용하는 sequence인 AATAAA는 poly(A)로부터 20 bp upstream에 위치하는 것을 확인하였다. 염기서열을 바탕으로 추정한 성장호르몬은 204개의 아미노산으로 구성된 단백질로서 signal peptide(17 aa)와 mature protein(187 aa)을 포함하고 있으며 mature protein의 분자량은 21.3 kDa으로 나타났다. 이 호르몬은 단백질의 3차 구조에 중요한 이황화 결합을 할 수 있는 4개의 cysteine 잔기와 1개의 N-glycosylation site를 가지고 있었다. 붉바리 성장호르몬의 염기서열은 농어목에 속하는 다른 어류의 성장호르몬 염기서열과의 유사도가 높게 나타났으며 orange-spotted grouper, gilthead seabream과 각각 96.9%, 88.6%의 상동성을 보였다.

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

  • 김경욱;한세진;노규섭
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제36권1호
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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.

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

  • 박현기;박재여;이향규
    • Journal of Korean Biological Nursing Science
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    • 제18권4호
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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.

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

  • 강상훈;김경준
    • 한방안이비인후피부과학회지
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    • 제26권1호
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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.

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

  • 김보민;한영은;이화진
    • 한국식품과학회지
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    • 제49권3호
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    • pp.331-337
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    • 2017
  • 배초향 추출물 및 분획별 생리활성 물질의 분포양상을 검정하기 위해 파이토케미컬 검색을 한 결과, 플라보노이드 확인시험에서 헥세인을 제외한 에틸아세테이트 및 부탄올 분획에서 양성반응을 확인하였다. 배초향 추출물의 극성에 따른 분획-헥세인, 에틸아세테이트 및 부탄올 분획의 산화방지 정도를 평가한 결과, 폴리페놀 함량, DPPH 라디칼 소거 및 환원력 실험에서 에틸아세테이트 분획이 다량의 폴리페놀을 함유하며 강력한 라디칼 소거 및 환원능력을 나타냈다. 또한 배초향 에틸아세테이트 분획의 항염 활성을 평가한 결과, 지방질다당류로 활성화된 BV-2 microglia에서 발생하는 과량의 산화질소(II) 생성을 억제하였을 뿐 아니라, 산화질소(II) 생성 효소인 iNOS 및 염증성 사이토카인 IL-6의 발현을 억제함을 확인하였다. 강력한 산화방지 및 항염증 활성을 나타내는 배초향 에틸아세테이트 분획내의 활성물질을 확인하고자 HPLC를 이용하여 분석한 결과, 배초향의 산화방지 물질로 잘 알려져 있는 로스마린산이 배초향 에틸아세테이트 분획에 함유되어 있음을 확인하였다.

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

  • 정성호;나민화;이재선;송상훈;윤을식;동은상;김한중
    • Archives of Plastic Surgery
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    • 제33권4호
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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.