• Title/Summary/Keyword: enhancer RNA

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Suppression of Lipid Accumulation in Differentiating 3T3-L1 Preadipocytes by a Standardized Commercial Juknyeok (죽력의 3T3-L1 지방전구세포에서 지방축적 억제 효과)

  • Jang, Byeong-Churl
    • Journal of Korean Medicine for Obesity Research
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    • v.22 no.1
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    • pp.38-46
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    • 2022
  • Objectives: Juknyeok (JN) is natural liquor extracted from bamboo stems (Phyllostachys bambusoides) and has been used as a traditional Korean medicine for improving vascular function, blood glucose, and treating stroke. Until now, the JN's lipid-lowering effect and underlying mechanism in adipocytes are poorly understood. The aim of this study was to scrutinize the effect of a standardized commercial JN on lipid accumulation during the differentiation of 3T3-L1 preadipocytes. Methods: Lipid and triglyceride (TG) accumulation in differentiating 3T3-L1 preadipocytes were measured by Oil Red O staining and AdipoRed assay, respectively. Cell count analysis was used to ascertain 3T3-L1 cytotoxicity. Immunoblotting and Reverse transcription polymerase chain reaction analysis were used to assess protein and messenger RNA (mRNA) expression levels in 3T3-L1 cells, respectively. Results: Treatment with JN at 25 𝜇l/ml after pH calibration with 6.35 significantly reduced lipid and TG accumulation in differentiating 3T3-L1 preadipocytes without significant cytotoxicity. On mechanistic levels, JN markedly suppressed protein expression levels of CCAAT/enhancer-binding protein (C/EBP)-𝛽 and fatty acid synthase (FAS) during the differentiation of 3T3-L1 preadipocytes. However, JN did not affect the protein expression levels of C/EBP-𝛼, peroxisome proliferator-activated receptor-𝛽/𝛾, and phosphorylation levels of signal transducer and activator of transcription-3/5 in differentiating 3T3-L1 preadipocytes. JN also reduced leptin mRNA expression levels in differentiating 3T3-L1 preadipocytes. Conclusions: JN at 25 𝜇l/ml lowers lipid accumulation and TG content in differentiating 3T3-L1 cells, mediated through the reduced expression levels of C/EBP-𝛽 and FAS.

Inhibition of Adipocyte Differentiation and Adipogenesis by Supercritical Fluid Extracts and Marc from Cinnamomum verum (초임계 추출 계피오일의 3T3-L1 지방전구세포의 분화 전사인자 억제에 의한 지방대사 조절)

  • Park, Sung-Jin;Lee, In-Seon;Lee, Sam-Pin;Yu, Mi-Hee
    • Journal of Life Science
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    • v.23 no.4
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    • pp.510-517
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    • 2013
  • This study was performed to evaluate the antiobesity effect of supercritical fluid extracts (SFC) and marc methanol extracts (SFM) from Cinnamomum verum in 3T3-L1 preadipocytes. In inducing the differentiation of 3T3-L1 preadipocytes in the presence of an adipogenic cocktail, iso-butylmethylanthine (IBMX), dexamathasone, and insulin, treatment with fraction residue SFC and SFM. SFC significantly reduced the mRNA expression of the transcription factor peroxisome proliferator-activate-dreceptor-${\gamma}$ ($PPAR{\gamma}$), the sterol regulatory-element-binding protein-1c (SREBP1c), and the CCAAT enhancer-binding-protein ${\alpha}$ ($C/EBP{\alpha}$) in a concentration-dependent manner. Moreover, SFC markedly down-regulated acyl-CoA synthetase-1 (ASC1), fatty acid synthesis (FAS), fatty acid transport-1 (FATP1), fatty acid binding protein 4 (FABP4), and perilipin. These findings suggest that SFC may be a potential therapeutic adjunct for obesity by targeting the differentiation of preadipocytes, as well as their functions.

A Review : On Exercise Performance Induction Gene Factors Change (운동이 유전자 조절물질에 미치는 영향에 관한 고찰)

  • Um, Ki-Mai;Yang, Yoon-Kwon;Kim, Tae-Woo
    • Journal of Korean Physical Therapy Science
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    • v.8 no.1
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    • pp.745-758
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    • 2001
  • The purpose of study to phenomenological examine and the mechanism regarding the gene(DNA, RNA, Protein) and sports to studied, analyzed. and evaluated. This review considers the evidence for genetic effects in several determinants of endurance performance and resistance performance, namely: body measurements and physique, body fat pulmonary functions, cardiac and circulatory functions, muscle characteristics. substrate utilization, maximal aerobic power and other. Moreover, the response to aerobic training of indicators aerobic work metabolism and endurance performance is reviewed, with emphasis on the specificity of the response and the individual differences observed in training ability. This study indicate that improvement of 'Enhancer Action' in RNA genes changed by exercise or sports. Moreover exercise was effect on Central Dogma with DNA makes RNA makes Protein. and think that occurred with exercise influence on skeletal muscle into cell have to Myosin Heavy Chain (MHC) changed was after exercise performance, which accompanied into skeletal muscle that were exercise-induces gene-modulation that is, take gene mutations. This study known that existed hormone(epinephrine)-immune system with interaction. Exercise were altered insulin binding and MAP Kinase signaling increased into immune cells. This review suggested that the high rate of glutamine utilization by cells of the immune system serves to maintain a high intra cellular concentration of the intermediates of biosynthetic pathways such that optimal rates of DNA, RNA and protein synthesis can be maintained. In the absence of glutamine, lymphocytes do not proliferate in vitro: proliferation increase greatly as the glutamine concentration increase. Glutamine is synthesized in skeletal muscle. Skeletal muscle and plasma glutamine levels are lowered by sepsis, injury, bums, surgery and endurance exercise and in the overtrained athlete. The study of result show that production of ET-1 is markedly increased tissue specifically in the heart by exercise without appreciable changes in endothelin-converting enzyme and endothelial receptor expressions, suggest that myocardial ET-1 may participate in modulation of cardiac function during exercise. Conclusionally, this study indicate that improvement of 'Enhancer Action' in RNA genes changed by exercise or sports. Moreover exercise was effect on Central Dogma with DNA makes RNA makes Protein. This study is expected to contribute the area of sports science, medicine, hereafter more effort is required to establish the relation between gene alters and exercise amount.

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Screening and functional validation of lipid metabolism-related lncRNA-46546 based on the transcriptome analysis of early embryonic muscle tissue in chicken

  • Ruonan, Chen;Kai, Liao;Herong, Liao;Li, Zhang;Haixuan, Zhao;Jie, Sun
    • Animal Bioscience
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    • v.36 no.2
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    • pp.175-190
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    • 2023
  • Objective: The study was conducted to screen differentially expressed long noncoding RNA (lncRNA) in chickens by high-throughput sequencing and explore its mechanism of action on intramuscular fat deposition. Methods: Herein, Rose crown and Cbb broiler chicken embryo breast and leg muscle lncRNA and mRNA expression profiles were constructed by RNA sequencing. A total of 96 and 42 differentially expressed lncRNAs were obtained in Rose crown vs Cobb broiler chicken breast and leg muscle, respectively. lncRNA-ENSGALT00000046546, with high interspecific variability and a potential regulatory role in lipid metabolism, and its predicted downstream target gene 1-acylglycerol-3-phosphate-O-acyltransferase 2 (AGPAT2), were selected for further study on the preadipocytes. Results: lncRNA-46546 overexpression in chicken preadipocyte 2 cells significantly increased (p<0.01) the expression levels of AGPAT2 and its downstream genes diacylglycerol acyltransferase 1 and diacylglycerol acyltransferase 2 and those of the fat metabolism-related genes peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α, fatty acid synthase, sterol regulatory element-binding transcription factor 1, and fatty acid binding protein 4. The lipid droplet concentration was higher in the overexpression group than in the control cells, and the triglyceride content in cells and medium was also significantly increased (p<0.01). Conclusion: This study preliminarily concludes that lncRNA-46546 may promote intramuscular fat deposition in chickens, laying a foundation for the study of lncRNAs in chicken early embryonic development and fat deposition.

Alteration of DNA Methylation in Oct-4 Gene in Mouse Preimplantation Embryos by the Interference RNA

  • Kim, Jong-Mu;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Chung, Hak-Jae;Chang, Won-Kyong;Kim, Nam-Hyung
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.21-28
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    • 2007
  • During early embryo development, Oct-4 is an important transcription factor for the early differentiation the present study was first examined methylation status in distal enhancer and promoter region of Oct-4 during mouse pre-implantation embryo development. In oocyte and sperm, high methylation was observed in both distal and proximal of promoter in Oct-4. Following fertilization relatively high methylation level remained until 8-cell stage embryos, but decreased at the morula and blastocyst stage. Specific gene knock down of Oct-4 by siRNA injection into zygote induced higher methylation rates of both distal and proximal region of promoter of Oct-4. These results suggest a functional link between the DNA methylation status of distal and promoter resign in the Oct-4 gene and the gene sequence-specific transcriptional silencing by exogenous siRNA injection during mouse preimplantation embryos.

Translational Enhancement by the 5' Leader of Tobacco Mosaic Virus and Soybean Glycinin Gene in Transgenic Tobacco Plants (담배 모자이크 바이러스와 대두 Glycinin 유전자의 5' Leader Sequence를 이용한 외래 유전자의 전이효율 증진)

  • Kang, Hong-Gu;Park, Jee-Won;Kim, Chung-Ho;Lim, Jae-Yun;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.38 no.3
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    • pp.224-231
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    • 1995
  • To increase the expression of a foreign protein in transgenic plant, the benefits of 5'-untranslated leader sequences of tobacco mosaic virus (TMV) RNA or soybean glycinin gene, Gy2, fused to a protein coding sequence were exploited. pGA643-derived plasmid contains 355 promoter of cauliflower mosaic virus, protein coding sequence of maize 10 kDa zein (10kZ) and Gy2 terminator. The leader from Gy2 or TMV RNA was inserted between the promoter and the coding sequence in each construct. The recombinant DNAs were introduced into tobacco plants by Agrobacterium mediated leaf disc transformation method. Although the transgene without the leader had more transcripts than the others, mRNAs containing the leader were translated more efficiently. It might be due to difference in the length of 5'-untranslated sequence and context surrounding the AUG codon, but could be sequence specific rather. These results suggest that the leader sequences of Gy2 and TMV play important roles as an enhancer in translational control of foreign gene in transgenic tobacco plant.

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Effects of Pueraria lobata Root Ethanol Extract on Adipogenesis and Lipogenesis During 3T3-L1 Differentiation into Adipocytes

  • Lee, Chae Myoung;Yoon, Mi Sook;Kim, Young Chul
    • Toxicological Research
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    • v.31 no.2
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    • pp.191-201
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    • 2015
  • We evaluated the inhibitory effect of Pueraria lobata root ethanol extract (PLREE) on lipid accumulation during 3T3-L1 differentiation to adipocytes by measuring the intracellular expression of adipogenic, lipogenic, and lipolytic markers and lipid accumulation. The total polyphenol and flavonoid content of PLREE were 47 and 29 mg/g, respectively. The electron donating capacity of PLREE at $1,000{\mu}g/mL$ was 48.8%. Treatment of 3T3-L1 preadipocytes with 100, 250, or $500{\mu}g/mL$ PLREE for 8 days dose-dependently promoted the differentiation of 3T3-L1 cells. In contrast, the lipid content of PLREE-treated cells was significantly reduced by 7.8% (p < 0.05), 35.6% (p < 0.001), and 42.2% (p < 0.001) following treatment with 100, 250, and $500{\mu}g/mL$ PLREE, respectively, as compared to differentiated control cells. PLREE upregulated peroxisome proliferator-activated receptor ${\gamma}$ mRNA and protein, and sterol regulator element-binding protein-1c mRNA levels, but did not affect CCAAT/enhancer binding-protein ${\beta}$ and ${\alpha}$ mRNA levels. PLREE also downregulated acetyl-CoA carboxylase mRNA and protein, fatty acid synthase (FAS) protein, and leptin mRNA levels, but did not affect FAS mRNA expression. PLREE upregulated adipose triglyceride lipase mRNA and protein expression, and hormone-sensitive lipase (HSL) protein expression, but did not affect HSL mRNA expression. In conclusion, we found that PLREE enhanced adipogenesis, but reduced lipogenesis, resulting in decreased lipid accumulation in 3T3-L1 cells.

Inhibition Effects of Galla Chinenisis Extract on Adipocyte Differentiation in OP9 Cells (오미자 추출물의 지방세포 분화 억제 효과)

  • Park, Sun-Young;Hwang, Hong-Yeon;Seo, Eun-A;Kwon, Kang-Beom;Ryu, Do-Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.4
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    • pp.455-461
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    • 2012
  • Obesity is associated with numerous diseases such as type 2 diabetes, hypertension and cancer. Inhibition of adipogenesis is a effectite strategy to anti-obesity. In this study, Galla Chinenisis extract (GCE) inhibited adipocyte differentiation in OP9 cells. There was no cytotoxicity when cells were treated with GCE in designated time intervals, unaffected by concentration. In this cell model, increases in fat storage were inhibited by 2 days treatment with various concentration of GCE, visualized by Oil red-O, BODIPY and DAPI staining. To understand the underlying mechanism at the molecular level, the effects of GCE were examined on the expression of the genes involved in adipogenesis by real-time PCR. In the progress of adipocyte differentiation with GCE-treated, the mRNA level of adipogenic genes such as peroxisome-proliferator-activated receptors gamma ($PPAR{\gamma}$), computer-assisted axial tomography/enhancer binding protein-alpha ($C/EBP{\alpha}$) were decreased. Also, GCE treatment inhibited increase of mRNA expression, which is adipogenic factor such as fatty acid synthase (FAS), hormone-sensitve lipase (HSL), lipoprotein lipase (LPL), and adipocyte-specific lipid binding protein (aP2). Therefore, the result of this study suggest that Galla Chinenisis extract can prevent adipocyte differentiation and GCE may have a great potential as a novel anti-adipogenic agent.

mRNA expression of myogenic-adipogenic makers and adipocyte in skeletal muscle of Hanwoo calves at newborn and 6 months of age

  • Ahn, Jun-Sang;Chung, Ki-Yong;Jang, Sun-Sick;Kim, Ui-Hyung;Hwang, So-Mi;Jin, Shil;Park, Bo-Hye;Kang, Dong-Hun;Kwon, Eung-Gi
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.893-902
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    • 2020
  • This study was conducted to compare the mRNA expression levels of myogenic-adipogenic makers in the skeletal muscle and adipocytes formation, body weight, rumen weight, and papilla length on Hanwoo calves at newborn and 6 months of age. Animals used three newborn Hanwoo calves (NC) and three Hanwoo calves 6 months of age (SC). Body weight and rumen weight were significantly increased in SC compared to NC (p < 0.01), and papilla length was longer about 10-fold in SC than NC. Adipocytes was possible to visually identify more adipocytes in SC compared to NC, and were mainly formed around the blood vessels. mRNA expression of myogenin, myosin heavy chain 1 and myosin heavy chain 2A in both longissimus dorsi (LD) and semimembranosus (SM) was found to increase with calves growth (p < 0.01), and it was confirmed that have higher levels of mRNA expression in SM than LD. In LD tissues, the mRNA expression of stearoyl-CoA desaturase (SCD, p < 0.03) and peroxisome proliferator activated receptor γ (PPARγ, p < 0.04) was significantly higher in SC than NC. In SM tissues, mRNA expression levels of SCD (p < 0.02) and CCAAT/enhancer binding protein β (C/EBPβ, p < 0.01) were higher in SC than NC, and also mRNA expression levels of PPARγ increased, but there was no significant difference. Thus, the calves period suggests that it is an important step in the development of the rumen and the myogenesis and adipogenesis.

An OTHBVS Cell Line Expresses the Human HBV Middle S Protein

  • Park, Sung-Gyoo;Guhung Jung
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.86-89
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    • 1999
  • An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.

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