• Reproductive and Developmental Biology
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• 제36권3호
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• pp.193-198
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• 2012

This study was to examine expression of the recombinant full-length adiponectin (recombinant adiponectin) in insect ovarian cell culture system and to characterize structural properties of the recombinant adiponectin secreted in medium. Gene construct encoding the recombinant adiponectin contained N-terminal collagen-like domain (110 Amino Acids, AAs), C-terminal globular domain (137 AAs) and C-terminal peptides for detection with V5 antibody (26 AAs included adaptor peptide) and purification using the 6xHis tag (6 AAs). The approximate molecular weight of the product (monomer) was 35 kDa. Molecular mass species of the expressed recombinant adiponectin were monomer (~35 kDa), dimer (~70 kDa), trimer (~105 kDa) and hexamer (~210 kDa). The major secreted species were the LMW forms, such as monomer, dimer, and trimer. There was MMW of hexamer as minor form. HMW multimers (~300 kDa) were shown as a tracer or not detected on the SDS-PAGE in several experiments (data not shown). The multimer forms in this study were not compatible to those in animal or human serum and adipose tissue by other researcher's study in which the major multimer forms were HMW. By protein denaturing experiments with reducing reagent (${\beta}$-MeOH), anionic detergent (SDS) and heat ($95^{\circ}C$) on the SDS-PAGE, not all adiponectin multimers seemed to have disulfide bond linked structure to form multimers. The recombinant adiponectin which expressed in insect ovarian cell culture system seemed to have the limitation as full physiological regulator for the application to animal and human study.

• 생명과학회지
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• 제24권8호
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• pp.895-902
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• 2014

아디포넥틴은 이미 합성된 GLUT4의 translocation 증가를 통해 포도당의 세포내 유입을 촉진하며 인슐린 민감도를 증가시키는 것으로 알려져 있다. 본 연구에서는 장기간(6주령부터 16, 26, 36, 47, 및 77주령까지)의 고지방식이(HFD)를 섭취한 비만 C57BL/6 생쥐와, 칼로리제한(CR) 또는 thiazolidinedione (TZD) 섭취에 의해 인슐린 민감성이 회복된 생쥐들로부터 지방조직을 적출하여 아디포넥틴과 GLUT4 의 mRNA 발현의 변화를 조사하였으며, 선형회귀분석(linear regression analysis)을 통해 아디포넥틴과 GLUT4 유전자 발현량 사이의 상관관계를 평가하여 아디포넥틴이 GLUT4 유전자 발현의 전사단계에서도 영향을 미치는지의 가능성을 확인하고자 하였다. 지방조직에서의 유전자 발현량은 TaqMan probe를 이용한 real-time PCR로 정량되었다. 실험결과, 지방조직에서의 아디포넥틴 mRNA발현량은 여러 조건의 생쥐 그룹들 사이에 유의한 변화가 나타나지 않았지만, GLUT4의 유전자 발현량은 HFD군에서는 감소하고, CR군(p<0.05)과 TZD군(p=0.007)에서는 유의하게 증가하는 변화가 확인되었다. 또한, 아디포넥틴과 GLUT4 mRNA 발현량 사이에는 유의한 상관관계를 나타내고 있음이 확인되었다. ND군(p<0.0001), HFD군 p<0.0001), 또는 각각의 주령과 식이별 소그룹, 그리고 CR군(p=0.002) 에서도 두 유전자간의 발현량이 유의하게 연관되어 있었다. 그러나 TZD군(p=0.73)의 생쥐에서는 그 연관성이 사라짐을 관찰하였다. 이는 TZD가 아디포넥틴 유전자 발현에는 영향을 미치지 않지만, GLUT4유전자 발현은 촉진하기에 두 유전자 사이에 유의하지 않은 상관관계로 변화되었음을 시사한다. 이들 결과는 아디포넥틴과 GLUT4의 유전자 발현은 강하게 연관되어 있으며, 두 유전자 발현 조절에 대한 공통적인 작용기전의 존재 가능성 또는 아디포넥틴이 GLUT4 translocation뿐만 아니라 GLUT4의 유전자 발현에도 직접적으로 작용하고 있음을 시사한다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권6호
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• pp.779-783
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• 2006

The adiponectin gene is known to be involved in the regulation of energy homeostasis involving food intake, carbohydrate and lipid metabolism. Human adiponectin gene polymorphisms have been recently reported to be associated with obesity, insulin sensitivity and the risk of type 2 diabetes. The present study was carried out to investigate the porcine adiponectin gene as a candidate gene for fat deposition and carcass traits. A mutation of A178G of the porcine adiponectin gene that resulted in substitution of the amino acid Isoleucine to Valine was identified. AcyI PCR-RFLP was used to detect the polymorphism of the genotypes in five different pig populations (Large White, Landrace, Duroc, Chinese breeds Meishan and Qingping). The A allele frequency was significantly higher among subjects from Chinsese lard type breeds, while the G allele was the only one present in those from Western lean type breeds. To determine if there was an association of the polymorphism with phenotypic variation, the mutation was tested in 267 pigs of the "Large $White{\times}Meishan$" F2 resource population. The results of association analyses showed significant associations of the genotypes with fat deposition and carcass traits. Allele G was significantly associated with increase in loin eye height, loin eye area and lean meat percentage and bone percentage, and decrease in fat mean percentage, ratio of lean to fat, shoulder fat thickness, 6-7 rib fat thickness, thorax-waist fat thickness and buttock fat thickness. The substitution of A178G (Ile60Val) happened to be located at amino acid 60 in the collagenous domain of porcine adiponectin which might affect the association into higher-order structures, and accordingly affect the posttranslational modifications and optimal biological activity of the multimeric forms. The identified functional polymorphism provides new evidence of adiponectin as an important candidate gene affecting fat deposition and carcass traits in pigs.

• Nutrition Research and Practice
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• 제1권4호
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• pp.363-370
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• 2007

Single nucleotide polymorphism (SNP) in adiponectin gene has been associated with insulin resistance, diabetes, and cardiovascular disease (CVD). This study was performed to investigate the association of SNP 276G${\rightarrow}$T at adiponectin gene with CVD risk factors in Korean type 2 diabetes mellitus (DM) patients. The subjects were 351 type 2 DM patients visited a DM clinic in Seoul, and the patients with known CVD were excluded. The adiponectin SNP 276G${\rightarrow}$T was analyzed and dietary intakes were assessed by a Food Frequency Questionnaire. The prevalence of G/G, G/T, and T/T genotype was 47.6%, 43.3%, and 9.1%, respectively. Male subjects with T/T genotype showed significantly lower level of adiponectin and HDL-cholesterol and significantly higher C-reactive protein (CRP) level compared to G/G and G/T genotypes. In G/G genotype, protein intake was negatively correlated to body weight, BMI, and waist circumference, and there were positive correlation between carbohydrate intake and BMI, waist-hip ratio, and ApoB/apoA-1 ratio in G/T genotype. However, in T/T genotype, there was no significant association between macronutrient intakes and anthropometric and hematological values. In conclusion, CVD risk would be high in type 2 DM patients with T/T genotype, and the association of macronutrient intakes with anthropometric and hematologic factors was different among the three adiponectin genotypes. These results may imply the need for different dietary management regime according to adiponectin genotype to lower CVD complications in Korean type 2 DM patients.

• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.588-593
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• 2004

The gene expression of porcine adiponectin and stearoyl coenzyme A desaturase (SCD) was investigated in this study. The partial gene sequences for adiponectin and SCD were amplified by RT-PCR from subcutaneous adipose tissue and cloned by TA cloning techniques. Sequences of these genes were determined and found to be highly homologous to that of other species, suggesting similar function of these genes as in other species. The transcripts of these adipocyte-related genes in pig tissues were measured by Northern analysis. The transcripts for adiponectin and SCD were highly expressed in porcine subcutaneous adipose tissue; the transcripts for SCD were also barely detected in the liver, but the greatest concentrations were in the adipose tissue. In porcine stromalvascular cells (S/V cells) cultured in vitro, transcripts for adiponectin and SCD increased gradually during adipocyte differentiation. The level of adipocyte adiponectin mRNA was associated with late adipocyte differentiation, indicating the gene may not be involved in adipocyte differentiation but has great importance in porcine adipocyte functions. The SCD transcripts were not detectable until 2 d after induction of adipocyte differentiation. It was highly expressed in differentiating porcine adipocytes (2 to 10 d after the induction of adipocyte differentiation), indicating a significant role of SCD in adipocytes.

• Clinical and Experimental Pediatrics
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• 제52권2호
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• pp.187-193
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• 2009

목 적 : Adiponectin은 대사 증후군에서 중요한 역할을 하는 물질이며, 비만, 2형 당뇨, 관동맥 질환 등에서 그 수치가 감소하는 것으로 알려져 있다. 고혈압과 혈청 adiponectin치의 관계는 여러 저자에 의하여 발표되었으나 아직 논란의 여지가 남아있는 상태이다. 우리는 낮은 adiponectin치가 청소년 고혈압과 어떤 관계가 있는지, 또 영향을 끼치는 기전이 유전적인지 혹은 생물학적인 것인지 알고자 유전자 다형성 분석을 하였다. 방 법 : 16에서 17세 사이의 40명의 고혈압 청소년과 20명의 대조군 청소년을 대상으로 각 군의 혈청 adiponectin, insulin, renin, aldosterone, ACE를 측정하여 비교하였다. cIMT와 PWV도 측정하였다. PCR법으로 adiponectin I164T 유전자 다형성을 조사하였다. 결 과 : 고혈압 청소년군에서 cIMT와 PWV가 의미있게 높았다. 혈청 insulin, renin, aldosterone치도 고혈압군에서 의미있게 높았다. 두 군간에 adiponectin치는 의미있는 차이를 보이지 않았다. 본 연구에 참여한 청소년에서는 TC genotype은 발견되지 않았고 모두 TT genotype을 보였다. 결 론 : 결론적으로 adiponectin치는 고혈압 청소년 군에서나 정상군에서나 차이가 없었다. 두 군에서 유전적인 다형성도 보이지 않았다. 더 큰 규모의 연구가 필요할 것으로 생각된다.

• 생명과학회지
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• 제19권10호
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• pp.1495-1498
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• 2009

Adiponectin은 지방세포에서 분비되는 아디포카인 중의 하나로서 에너지 대사, 인슐린 저항성, 심혈관계에 조절역할을 하는 것으로 밝혀지고 있다. Adiponectin 유전자는 염색체상에서 제2형 당뇨병 감수성과 연관된 것으로 확인된 3q27 부위에 위치하는데[4], adiponectin 유전자의 일부 유전자 다형성이 adiponectin 농도, 제2형 당뇨병, 비만과 연관되어 있음이 보고된 바 있으며, 그 중 대표적인 것이 T45G와 G276T이다. 따라서 본 연구에서는 adiponectin 유전자 다형성과 제2형 당뇨병 사이에 어떠한 연관성이 한국인에서도 있는지 알아보았다. 연구 결과 T45G 유전자 다형성은 제2형 당뇨병과 유의성을 가짐을 확인하였으나, G276T의 경우에는 유의적인 연관성을 보이지 않는 것으로 확인되었다. 이는 여러 인구 집단과 비교하였을 때에 T45G 유전자 다형성의 경우에는 한국인에서 제2형 당뇨병의 marker로 사용하는 것이 가능하다고 판단된다. Adiponectin은 이미 항염증반응, 항동맥경화 작용이 있다는 것이 알려져 있을 뿐만 아니라 제2형 당뇨병과도 관련이 있다고 보고되어 있으므로 adiponectin을 증가시킴으로써 제2형 당뇨병이나 비만 등의 치료 및 예방 효과를 보일 수 있을 것으로 보고 있다. 따라서 adiponectin의 분자 수준에서의 연구를 위해서 유전자 다형성과 제2형 당뇨병과의 연관성에 대한 연구는 필수적이라 할 수 있으며, adiponectin 유전자 다형성에 관한 본 연구결과의 임상적인 의의를 확실하게 확인하기 위하여서는 향후 대상 환자 수와 대조군을 더욱 늘리고 다양한 adiponectin 유전자 다형성을 후보로 하여 추가적인 연구를 할 필요가 있을 것으로 생각된다.

• IMMUNE NETWORK
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• 제6권1호
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• pp.6-12
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• 2006

Background: The Hypothalamo-Pituitary-Adrenal (HPA) axis is an important regulator for the body's stress response. As a primary stress responsive system, HPA-axis secretes various neurotransmitters, hormones, and cytokines, which regulates the immune system. Natural killer (NK) cell which is plays an important role in the innate immune response, is specially decreased their numbers and loose cytolytic activity in response to stress. However, the effect of HPA-axis secreted proteins on NK cell activity has not been defined. Herein, we studied the effect of adrenal secreted adiponectin on NK cell cytotoxicity. Adiponectin which is well-known metabolic control protein, plays important roles in various diseases, including hypertension, cardiovascular diseases, inflammatory disorders, and cancer. Methods: Signal sequence trap was used to find stress novel secretory protein from HP A-axis. Selected adiponectin was treated mouse mature primary NK cells and then examined the effect of adiponectin to NK cell cytotoxicity and cytokine expression level. Results: We found that adiponectin which is secreted from adrenal gland, suppress IL-2 induced NK cell cytotoxicity. And also investigated cytolytic cytokines are suppressed by adiponectin. Conclusion: These data suggest that adiponectin inhibites NK cell cytotoxicity via suppression of cytotoxicity related target gene.

• Asian-Australasian Journal of Animal Sciences
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• 제25권9호
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• pp.1316-1321
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• 2012

Adipokines, adipocyte-derived protein, have important roles in various kinds of physiology including energy homeostasis. Chemerin, one of adipocyte-derived adipokines, is highly expressed in differentiated adipocytes and is known to induce macrophage chemotaxis and glucose intolerance. The objective of the present study was to investigate the changes of chemerin and the chemokine-like-receptor 1 (CMKLR1) gene expression levels during differentiation of the bovine adipocyte and in differentiated adipocytes treated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), adiponectin, leptin, and chemerin (peptide analog). The expression levels of the chemerin gene increased at d 6 and 12 of the differentiation period accompanied by increased cytoplasm lipid droplets. From d 6 onward, peroxisome proliferator-activated receptor-${\gamma}2$ (PPAR-${\gamma}2$) gene expression levels were significantly higher than that of d 0 and 3. In contrast, CMKLR1 expression levels decreased at the end of the differentiation period. In fully differentiated adipocytes (i.e. at d 12), the treatment of TNF-${\alpha}$ and adiponectin upregulated both chemerin and CMKLR1 gene expression levels, although leptin did not show such effects. Moreover, chemerin analog treatment was shown to upregulate chemerin gene expression levels regardless of doses. These results suggest that the expression of chemerin in bovine adipocyte might be regulated by chemerin itself and other adipokines, which indicates its possible role in modulating the adipokine secretions in adipose tissues.

• Biomolecules & Therapeutics
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• 제26권5호
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• pp.446-457
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• 2018

Adiponectin, a hormone predominantly originated from adipose tissue, has exhibited potent anti-inflammatory properties. Accumulating evidence suggests that autophagy induction plays a crucial role in anti-inflammatory responses by adiponectin. However, underlying molecular mechanisms are still largely unknown. Association of Bcl-2 with Beclin-1, an autophagy activating protein, prevents autophagy induction. We have previously shown that adiponectin-induced autophagy activation is mediated through inhibition of interaction between Bcl-2 and Beclin-1. In the present study, we examined the molecular mechanisms by which adiponectin modulates association of Bcl-2 and Beclin-1 in macrophages. Herein, we demonstrated that globular adiponectin (gAcrp) induced increase in the expression of AUF1 and ZFP36L1, which act as mRNA destabilizing proteins, both in RAW 264.7 macrophages and primary peritoneal macrophages. In addition, gene silencing of AUF1 and ZFP36L1 caused restoration of decrease in Bcl-2 expression and Bcl-2 mRNA half-life by gAcrp, indicating crucial roles of AUF1 and ZFP36L1 induction in Bcl-2 mRNA destabilization by gAcrp. Moreover, knock-down of AUF1 and ZFP36L1 enhanced interaction of Bcl-2 with Beclin-1, and subsequently prevented gAcrp-induced autophagy activation, suggesting that AUF1 and ZFP36L1 induction mediates gAcrp-induced autophagy activation via Bcl-2 mRNA destabilization. Furthermore, suppressive effects of gAcrp on LPS-stimulated inflammatory mediators expression were prevented by gene silencing of AUF1 and ZFP36L1 in macrophages. Taken together, these results suggest that AUF1 and ZFP36L1 induction critically contributes to autophagy induction by gAcrp and are promising targets for anti-inflammatory responses by gAcrp.