• Title/Summary/Keyword: insulin binding

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Identification of intrinsic regulators in the secondary palate morphogenesis (이차구개 형태분화의 내적 조절유전자 규명)

  • Lee, Jae-Guk;Jang, Eun-Ha;Im, Yang-Hee;Kim, Ki-Byeung;Ko, Seung-O;Cho, Eui-Sic;Shin, Hyo-Keun
    • Korean Journal of Cleft Lip And Palate
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    • v.10 no.1
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    • pp.1-16
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    • 2007
  • 이차구개는 발생과정에서 구개선반의 형성과 성장, 거상과 융합의 과정을 통해 형성된다. 이와 같은 이차구개의 형성과정은 미세한 분자유전학적 신호전달기전에 의해 조절되는 것으로 알려져 있어서, 신호전달과정에 관여하는 유전인자의 발현이상이 되면 정상적인 이차구개가 형성되지 못하고 구개파열이라는 선천성 기형이 발생된다. 구개파열의 유발인자들에 대한 많은 연구에도 불구하고 현재까지 정상적인 이차구개의 형성을 조절하는 분자유전학적 기전에 대해서는 명확히 알려져 있지 않다. 따라서 본 연구에서는 이차구개의 형태분화를 조절하는 분자유전학적 기전을 알아보고자, 이차구개 형성의 내적 조절인자 중 핵심유전자로 알려져 있는 Osr2가 결손된 생쥐의 이차구개 형성과정에서 정상생쥐에 비해 발현의 변동이 나타나는 유전자를 확인하였다. 유전자 발현의 변동은 발생 14.5일(E14.5)의 구개선반으로부터 추출한 total RNA를 이용하여 ACP-based GeneFishing PCR을 시행하여 확인하였고, 각각의 변동된 유전자를 동정하여 정상생쥐의 이차구개 형성과정에서의 발현양상을 in situ hybridization을 시행하여 확인하였다. 총 120쌍의 primer를 이용한 검색을 통해서 정상생쥐의 구개선반에 비해 mutant에서 발현이 변동된 유전자는 7개가 검출되었고, 이들은 모두 정상생쥐에 비해 mutant에서 발현이 증가되는 것으로 확인되었다. 검출된 유전자는 vimentin(Vim), ${\beta}$-tropomyosin 2(Tpm2), thioredoxin-like 5(Txnl5), procollagen type II alpha 1(Col2a1), Insulin-like growth factor binding protein 7(IGFbp7), Sui 1 homologs(Sui 1), Defender against cell death1(Dad1)이었다. 검출된 유전자를 동정하여 정상생쥐의 구개 형성과정에서의 발현양상을 알아본 결과, Col2a1 을 제외한 유전자들은 모두 E13.5의 구개선반에서 특이적으로 발현되고 있었으나 구개선반이 융합된 E15.5에서는 Vim, Txnl5 그리고 Dad1 만이 봉합선을 따라 발현이 지속되고 있었다. 이상의 결과로 보아 검출된 유전자들은 구개선반의 형태분화과정에서 발현되어 이차구개의 형성과정에 관여할 것으로 여겨진다. 또한 이들은 이차구개 형성의 내적조절인자인 Osr2의 downstream target 으로 구개선반의 성장과 융합과정에 직접적으로 관여하는 유전물질일 것으로 추정된다.

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Effects of the KI-180 and KI-188 Calcium Formulae on the Growth and Development of Rats (KI-180과 KI-188 칼슘조성물이 어린쥐의 성장발육에 미치는 영향)

  • Park, Seung-Man;Seong, Ki-Seung;Lee, Jong-Seok;Lee, Ok-Hwan;Han, Chan-Kyu
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.358-363
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    • 2014
  • Herbs have active components that promote the growth rate of both animals and human. The KI-180 and KI-188 calcium food formulae contain Acanthopanacis cortex, Bombysis corpus and hoelen, seaweed calcium, chlorella extract, spirulina, colostrum powder, and other natural and functional components. We evaluated the growth-promoting effects of these formulae by analyzing the weight, femur and backbone, alkaline phosphatase, osteocalcin, testosterone, insulin-like growth factor (IGF)-1, and IGF binding protein-3 (IGFBP-3) of growing rats. Growing rats administered with KI-180 and KI-188 calcium showed the increase of body weight, body length, and femur weight and length of growing rats. In addition, administration of KI-180 and KI-188 calcium increased the alkaline phosphatase activity, the levels of osteocalcin and the growth hormones IGF-1 and IGFBP-3 of growing rats. The impact of KI-180 and KI-188 calcium on the physical development of growing rats suggests that the incorporation of these food formulae in the diets of growing children may promote the physical development.

Inhibitory Effect of Rumex Crispus L. Fraction on Adipocyte Differentiation in 3T3-L1 Cells (소리쟁이 분획물의 지방세포 분화 억제 효과)

  • Park, Sung-Jin;Choi, Jun-Hyeok;Jung, Yeon-Seop;Yu, Mi Hee
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.90-96
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    • 2013
  • The anti-obesity effect of ethanol xtract and their fractions from Rumex Crispus L. on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil red O assay, western blot and real-time PCR analysis. Ethyl acetate fraction of Rumex crispus L. significantly inhibited adipocyte differentiation when treated during the adipocyte differentiation process, as assessed by measuring fat accumulation using Oil red O staining. In inducing differentiation of 3T3-L1 preadipocytes in the presence of an adipogenic cocktail, isobutylmethylxanthine (IBMX), dexamethasone- and insulin-along with ethyl acetate fraction residue processing treatment significantly decreased protein expression of obesity-related proteins, such as peroxisome-proliferators-activated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). These results indicate that ethyl acetate fraction of Rumex crispus L. is the most effective candidate for preventing obesity. However further studies will be needed to identify the active compounds that confer the anti-obesity activity of ethyl acetate fraction from Rumex crispus L.

Effects of Ethanol Extract of Sargassum horneri on Adipocyte Differentiation and Adipogenesis in 3T3-L1 Preadipocytes (괭생이모자반 에탄올 추출물이 3T3-L1 지방전구세포의 분화 및 adipogenesis에 미치는 영향)

  • Kwon, Da Hye;Choi, Yung Hyun;Kim, Byung Woo;Hwang, Hye Jin
    • Journal of Life Science
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    • v.29 no.2
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    • pp.209-214
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    • 2019
  • Sargassum horneri (Turner) C. Agardh is a marine brown algae widely distributed in the North Pacific Ocean. It is known for its anti-inflammatory and anti-atopic effects. In this study, we determined the effects of ethanol extract of Sargassum horneri (Turner) C. Agardh (EESH) on anti-obesity activities in 3T3-L1 preadipocytes. Our results indicated that treatment with EESH decreased the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in lipid droplet content observed by oil red O staining. The concentrations of cellular triglycerides were also reduced in 3T3-L1 cells after treatment with EESH. Triglyceride content was inhibited by 13%, 16%, and 23% after treatment with 250, 500, and $1,000{\mu}g/ml$ of EESH in 3T3-L1 cells, respectively. Western blotting analysis showed that EESH suppressed adipogenic transcription factor expression in a dose dependent manner. Specifically, it suppressed cytidine-cytidine-adinosine-adenosine-thymidine (CCAAT) /enhancer binding proteins $(C/EBP){\alpha}$, $C/EBP{\beta}$ and peroxisome proliferator-activated receptor $(PPAR){\gamma}$. This indicated that EESH could control the expression of adipogenic transcription factors and inhibit the differentiation of adipocytes. Taken together, these findings demonstrated that EESH showed anti-obesity effects and could have potential uses in the field of nutraceuticals.

Analysis of Pregnancy-Associated Plasma Protein-A (PAPP-A) in Porcine Neonatal Testis (미성숙 돼지 정소 내에서의 pregnancy-associated plasma protein-A 특성 분석)

  • Lee, W.Y.;Cho, K.H.;Yeo, J.M.;Shin, Y.K.;Park, J.K.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.1
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    • pp.5-13
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    • 2020
  • The identify of biomarkers in living tissues is useful to understand the characteristics and functions of the cells. Proteins such as protein gene product 9.5, promyelocytic leukemia zinc finger, NANOG, and stage-specific embryonic antigen-1 have been identified as markers for porcine undifferentiated spermatogonia. In this study, the expression of insulin-like growth factor binding proteins (IGFBPs), a newly discovered porcine spermatogonia marker and pregnancy-associated plasma protein-A (PAPP-A), a protein regulator of IGFBPs, were characterized in 5-day-old porcine testis. To analyze the function of IGFBPs, RT-PCR was performed. IGFBP 2, 3, 4, and 6 were detected in porcine spermatogonia and PAPP-A was detected in basement regions in 5day old porcine seminiferous tubules. PAPP-A was not expressed in spermatogonia, but it was expressed in Sertoli cells. These results suggest that the expression of PAPP-A protein in Sertoli cells may regulate the development and differentiation of testicular cells through the IGF axis in porcine neonatal testis.

Effects of protein content and the inclusion of protein sources with different amino acid release dynamics on the nitrogen utilization of weaned piglets

  • Hu, Nianzhi;Shen, Zhiwen;Pan, Li;Qin, Guixin;Zhao, Yuan;Bao, Nan
    • Animal Bioscience
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    • v.35 no.2
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    • pp.260-271
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    • 2022
  • Objective: We aimed to investigate the effect of the differing amino acid (AA) release dynamics of two protein sources on the growth performance, nitrogen deposition, plasma biochemical parameters, and muscle synthesis and degradation of piglets when included in their diets at normal and low concentrations. Methods: Forty-eight piglets (Duroc×Landrace×Large White) with initial body weight of 7.45±0.58 kg were assigned to six groups and fed one of 6 diets. The 6 dietary treatments were arranged by 3×2 factorial with 3 protein sources and 2 dietary protein levels. They are NCAS (a normal protein content with casein), NBlend (a normal protein content with blend of casein and corn gluten meal), NCGM (a normal protein content with corn gluten meal), LCAS (a low protein content with casein), LBlend (a low protein content with blend of casein and corn gluten meal), LCGM (a low protein content with corn gluten meal). The release dynamics of AA in these diets were determined by in vitro digestion. The digestibility, utilization and biological value of nitrogen in piglets were determined by micro Kjeldahl method. Plasma insulin was measured by enzyme-linked immunosorbent assay kits. The protein expression of mediators of muscle synthesis and degradation was determined by western blotting. Results: Although the consumption of a low-protein diet supplemented with crystalline AA was associated with greater nitrogen digestion and utilization (p<0.05), the final body weight, growth performance, nitrogen deposition, and phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein 1 in the muscle of pigs in the low-protein diet-fed groups were lower than those of the normal-protein diet-fed groups (p<0.05) because of the absence of non-essential AA. Because of the more balanced release of AA, the casein (CAS) and Blend-fed groups showed superior growth performance, final body weight and nitrogen deposition, and lower expression of muscle ring finger 1 and muscle atrophy F-box than the CGM-fed groups (p<0.05). Conclusion: We conclude that the balanced release of AA from CAS containing diets and mixed diets could reduce muscle degradation, favor nitrogen retention, % intake and improve growth performance in pigs consuming either a normal- or low-protein diet.

Effect of Antibody Immobilization Method to Magnetic Micro Beads on its Immunobinding Characteristics (자성 미세입자에의 항체 고정화 방법이 면역결합반응에 미치는 영향)

  • Choi, Hyo Jin;Hwang, Sang Youn;Jang, Dae Ho;Cho, Hyung Min;Kang, Jung Hye;Seong, Gi Hun;Choo, Jae Bum;Lee, Eun Kyu
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.65-72
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    • 2006
  • Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

Inhibition of Adipocyte Differentiation and Adipogenesis by Aged Black Garlic Extracts in 3T3-L1 Preadipocytes (흑마늘 추출물에 의한 3T3-L1 지방전구세포의 분화 및 adipogenesis 억제에 관한 연구)

  • Park, Jung-Ae;Park, Cheol;Han, Min-Ho;Kim, Byung-Woo;Chung, Yoon-Ho;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.21 no.5
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    • pp.720-728
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    • 2011
  • Garlic (Allium sativum) has been used as a source food as well as a traditional folk medicine ingredient since ancient times. Aged black garlic is a type of fermented garlic and is expected to have stronger anticancer and antioxidant activities than raw garlic. However, the mechanisms of their inhibitory effects on adipocyte differentiation and adipogenesis are poorly understood. In the present study, the effects and mechanisms of water extracts of raw garlic (WERG) and aged black garlic (WEABG) on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes were investigated. Treatment with WEABG significantly suppressed terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner as confirmed by a decrease in lipid droplet number and lipid content through Oil Red O staining, however WERG had no such effect. In addition, WEABG reduced accumulation of cellular triglyceride, which is associated with a significant inhibition of key pro-adipogenic transcription factors including peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ (C/EBP${\alpha}$) and C/EBP${\beta}$. Taken together, these results provide important new insight that aged black garlic might inhibit adipogenesis by suppressing the pro-adipogenic transcription factors in 3T3-L1 preadipocytes, and further studies will be needed to identify the active compounds that confer the anti-obesity activity of aged black garlic.

Effects of Myelophycus Simplex Papenfuss Methanol Extract on Adipocyte Differentiation and Adipogenesis in 3T3-L1 Preadipocytes (바위수염 메탄올 추출물이 3T3-L1 지방전구세포의 분화에 미치는 영향)

  • Kim, Hyang Suk;Kwon, Da Hye;Cheon, Ji Min;Choi, Eun Ok;Kim, Ji Hyun;Han, Min Ho;Choi, Yung Hyun;Kim, Byung Woo;Hwang, Hye Jin
    • Journal of Life Science
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    • v.25 no.1
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    • pp.62-67
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    • 2015
  • Myelophycus simplex Papenfuss is distributed over the northern Pacific and southern coast of Korea, and is a member of the brown algae family. The objective of this study was to investigate the effect of M. simplex methanol extract on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes. Treatment with M. simplex methanol extract significantly suppressed terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in lipid droplet content observed by Oil Red O staining. Also, the M. simplex methanol extract significantly suppressed the triglyceride content of 3T3-L1 preadipocytes in a dose-dependent manner. Treatment with 300 and $500{\mu}g/ml$ of M. simplex methanol extract caused a 42% and 76% reduction in lipid droplet content, respectively. In order to understand the anti-adipogenic effects of M. simplex methanol extract, the changes in the expression of several adipogenic transcription factors, including peroxisome proliferator-activated receptor (PPAR) ${\gamma}$-cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding protein (C/EBP) ${\alpha}$ and ${\beta}$, were investigated using immunoblotting. M. simplex suppressed the expression of $PPAR{\gamma}$, $C/EBP{\alpha}$, and $C/EBP{\beta}$ proteins compared with control. Therefore, the results of this study suggest that M. simplex methanol extract inhibits adipocyte differentiation and thus may have applications as a potential source for an anti-obesity functional food agent.

Effects of Garcinia cambogia Extract on the Adipogenic Differentiation and Lipotoxicity (가르시니아 캄보지아 추출물의 지방세포 분화 및 지방 독성에 미치는 영향)

  • Kang, Eun Sil;Ham, Sun Ah;Hwang, Jung Seok;Lee, Chang-Kwon;Seo, Han Geuk
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.411-416
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    • 2013
  • This study aimed to examine the mechanisms underlying the effects of Garcinia cambogia extract on the adipogenic differentiation of 3T3-L1 cells and long-chain saturated fatty acid-induced lipotoxicity of HepG2 cells. 3T3-L1 preadipocytes, mouse embryonic fibroblast-adipose like cell line, were treated with MDI solution (0.5 mM IBMX, 1 ${\mu}M$ dexamethasone, 10 ${\mu}g/mL$ insulin) to generate a cellular model of adipocyte differentiation. Using this cellular model, the anti-obesity effect of Garcinia cambogia extract was evaluated. MDI-induced lipid accumulation and expression of adipogenesis-related genes were detected by Oil red O staining, Nile Red staining, and Western blot analysis. Effects Garcinia cambogia extract on palmitate-induced lipotoxicity was also analyzed by MTT assay, LDH release, and DAPI staining in HepG2 cells. Garcinia cambogia extract significantly suppressed the adipogenic differentiation of preadipocytes and intracellular lipid accumulation in the differentiating adipocytes. Garcinia cambogia extract also markedly inhibited the expression of peroxisome proliferator- activated receptor ${\gamma}2$ ($PPAR{\gamma}2$), CCAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), and adipocyte protein aP2 (aP2). In addition, Garcinia cambogia extract significantly attenuated palmitate-induced lipotoxicity in HepG2 cells. Palmitateinduced cellular damage and reactive aldehydes were also significantly reduced in the presence of Garcinia cambogia extract. These findings suggest that the Garcinia cambogia extract inhibits the adipogenic differentiation of 3T3-L1 preadipocytes, probably by regulating the expression of multiple genes associated with adipogenesis such as $PPAR{\gamma}2$, $C/EBP{\alpha}$, aP2, and thereby modulating fatty acid-induced lipotoxicity to reduce cellular injury in hepatocytes.