• Journal of Veterinary Science
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• 제25권2호
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• pp.31.1-31.8
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• 2024

Background: Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy. Objectives: This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype. Methods: Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry. Results: fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 ^-, CD34 ^-, CD45 ^-, CD9⁺, and CD44⁺. Conclusions: In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

• Nutrition Research and Practice
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• 제9권6호
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• pp.599-605
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• 2015

BACKGROUND/OBJECTIVES: Citrus flavonoids have a variety of physiological properties such as anti-oxidant, anti-inflammation, anti-cancer, and anti-obesity. We investigated whether bioconversion of Citrus unshiu with cytolase (CU-C) ameliorates the anti-adipogenic effects by modulation of adipocyte differentiation and lipid metabolism in 3T3-L1 cells. MATERIALS/METHODS: Glycoside forms of Citrus unshiu (CU) were converted into aglycoside forms with cytolase treatment. Cell viability of CU and CU-C was measured at various concentrations in 3T3L-1 cells. The anti-adipogenic and lipolytic effects were examined using Oil red O staining and free glycerol assay, respectively. We performed real time-polymerase chain reaction and western immunoblotting assay to detect mRNA and protein expression of adipogenic transcription factors, respectively. RESULTS: Treatment with cytolase decreased flavanone rutinoside forms (narirutin and hesperidin) and instead, increased flavanone aglycoside forms (naringenin and hesperetin). During adipocyte differentiation, 3T3-L1 cells were treated with CU or CU-C at a dose of 0.5 mg/ml. Adipocyte differentiation was inhibited in CU-C group, but not in CU group. CU-C markedly suppressed the insulin-induced protein expression of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) as well as the mRNA levels of $CEBP{\alpha}$, $PPAR{\gamma}$, and sterol regulatory element binding protein 1c (SREBP1c). Both CU and CU-C groups significantly increased the adipolytic activity with the higher release of free glycerol than those of control group in differentiated 3T3-L1 adipocytes. CU-C is particularly superior in suppression of adipogenesis, whereas CU-C has similar effect to CU on stimulation of lipolysis. CONCLUSIONS: These results suggest that bioconversion of Citrus unshiu peel extracts with cytolase enhances aglycoside flavonoids and improves the anti-adipogenic metabolism via both inhibition of key adipogenic transcription factors and induction of adipolytic activity.

• Journal of Nutrition and Health
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• 제52권1호
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• pp.17-25
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• 2019

본 연구에서는 댕댕이나무 열매 추출물이 3T3-L1과 마우스 지방유래줄기세포의 지방 분화유도 및 지방생성에 미치는 영향을 살펴보았다. 3T3-L1에 댕댕이나무 열매 추출물을 처리하였을 때, 농도의존적으로 지방구의 생성을 줄였고 지방세포 분화에 있어서 중요한 전사인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$, SREBP1의 발현을 억제시켜 지방 합성이 감소됨을 확인하였다. 또한, 마우스 지방에서 분리한 줄기세포의 지방 분화과정에서도 댕댕이나무 열매 추출물이 $PPAR{\gamma}$, $C/EBP{\alpha}$, SREBP1의 단백질 발현을 감소시켜 지방 축적을 농도 의존적으로 억제하였다. 이상의 결과로 댕댕이나무 열매 추출물은 세포독성이 없는 농도에서 지방 세포의 분화를 억제 하는 것으로 확인되어 항비만 기능성 소재로서의 활용 가능성이 있을 것으로 사료된다.

• 한국컴퓨터정보학회논문지
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• 제29권1호
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• pp.187-194
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• 2024

본 연구에서는 산딸기 잎의 항비만 기능성 소재로서의 가능성을 확인하기 위하여 3T3-L1 지방전구세포에 산딸기 잎 추출물을 처리한 후 중성지방 함량을 측정하고 지방생성과 관련된 여러 단백질의 발현 양상을 측정하였다. 산딸기 잎은 열수로 추출·농축하여 시료로 사용하였으며 추출물의 수율은 4.76%였다. 산딸기 잎 추출물에 의한 3T3-L1 세포 생존율을 측정한 결과, 1,000 ㎍/mL 농도에서 13%의 성장 저해를 확인할 수 있었다. 이에 본 연구에서 시료 처리 농도를 500 ㎍/mL까지 처리하여 실험을 진행하였다. 세포 내 중성지방의 생성은 농도의존적으로 감소하는 경향을 나타내었으며, 감소율은 대조군에 비하여 농도 100, 300, 500 ㎍/mL에서 각각 28.7, 40.8, 51.6%를 나타내었다. 또한 PPARγ, C/EBPα 및 SREBP-1c, FAS 및 ACC의 발현 억제, p-AMPK 발현 증가로 지방합성이 억제되는 것을 확인하였다. 이상의 결과, 산딸기 잎 추출물이 지방세포 분화 및 지방생성 관련 인자들을 조절함으로서 비만 개선 소재로서의 활용이 가능할 것으로 판단된다.

• Molecules and Cells
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• 제24권3호
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• pp.388-393
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• 2007

Osteonecrosis of the femoral head (ONFH) is a multifactorial disease to which certain individuals are more at risk. Altered lipid metabolism is one of the major risk factors for osteonecrosis, especially corticosteroid therapy and alcoholism. Peroxisome Proliferator-Activated Receptor-${\gamma}$ ($PPAR{\gamma}$) plays a crucial role in differentiation of mesenchymal cells to adipocytes, lipid homeostasis, and bone metabolism. To investigate the possible association between $PPAR{\gamma}$ gene variants and susceptibility to ONFH, we genotyped three common polymorphisms (-796A > G, +34C > G[Pro12Ala], and +82466C > T[His477His]) in 448 ONFH patients and 336 control subjects. Genotypes, allele frequencies, and haplotypes of the polymorphisms in the complete set of patients as well as in subgroups by sex or etiology were not significantly different from those in the control group. This suggests that the examined polymorphisms and haplotypes of the $PPAR{\gamma}$ gene are unlikely to be associated with susceptibility to ONFH.

• Journal of Periodontal and Implant Science
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• 제37권3호
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• pp.479-488
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• 2007

In spite of the attention given to the study of mesenchymal stem cells derived periodontal ligament (PDL), there is a lack of information about canine PDL cells. In this study, we characterized canine PDL cells to clarify their stem cell properties, including self renewal, proliferate rate, stem cell markers and multipotency. PDL cells were obtained from extracted premolars of canines, following a colony forming assay and proliferation rate of sub-confluent cultures of cells for self-renewal, immunostaining for STRO-1 and CD146/MUC18 and a differentiation assay for multipotency. Canine PDL cells formed single-cells colonies and 25% of the PDL cells displayed positive staining for BrdU. The cells expressed the mesenchymal stem-cell markers, STRO-1 and CD146/MUC18. Under defined culture conditions, the cells differentiated into osteoblasts and adipocytes, but the cells didn't differentiated into chondrocytes. The findings of this study indicated that the canine PDL cells possess crucial stem cells properties, such as self-renewal and multipotency, and express the mesenchymal stem cell markers on their surface. The isolation and characterization of canine PDL cells makes it feasible to pursue preclinical models of periodontal regeneration in canine.

• 한국식품영양학회지
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• 제30권6호
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• pp.1292-1298
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• 2017

The purpose of this study was to evaluate the antioxidant and anti-adipogenic activities of Ligularia stenocephala (L. stenocephala) extract. The contents of the total polyphenol of the extract was 55.950 mg GAE/g residue. Antioxidant activities of L. stenocephala were evaluated by free radical scavenging ability and a reducing power test. 2,2'azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ${\alpha}$-${\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) free radical scavenging activities of the extract were approximately 90% and 70%, respectively. Reducing power of the extract was 258.833 mg TE/g residue. The anti-adipogenic activity of L. stenocephala extract was examined in 3T3-L1 cells. During adipocyte differentiation, the 3T3-L1 cells were treated both with and without the extract. L. stenocephala extract suppressed the lipid accumulation in a concentration-dependent manner in the 3T3-L1 cells. The L. stenocephala extract inhibited the expression of peroxisome proliferator activated receptor ${\gamma}$ ($PPAR{\gamma}$) and adipocyte protein 2 (aP2) proteins, compared with control adipocytes. These results indicate that L. stenocephala could be regarded as a potential source natural antioxidant and an anti-obesity agent.

• 생약학회지
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• 제42권4호
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• pp.302-308
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• 2011

As a part of our ongoing program on finding biologically active components from natural source we found three known constituents from the EtOH extract of the wheat bran. The known compounds were identified as tachioside (1), pinellic acid (2) and tryptophan (3). The structure and relative stereochemistry were determined from MS, 1D and extensive 2D NMR techniques as well as by comparison of their data with the published values. All isolates were tested their inhibitory effects on the adipogenesis in 3T3-L1 cells. The effect of compounds from wheat bran on 3T3-L1 adipocyte differentiation were measured by Oil Red O staining. These results demonstrate that tachioside (1) and pinellic acid (2) decreased lipid content in 3T3-L1 adipocytes by inhibiting lipogenesis. These compounds had shown antiobesity activities.

• Natural Product Sciences
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• 제17권4호
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• pp.273-278
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• 2011

In this study, the effects of a methanol (MeOH) extract of Carduus crispus L. (Asteraceae) on adipogenesis was investigated in 3T3-L1 cells. To differentiate preadipocytes to adipocytes, confluent 3T3-L1 preadipocytes were treated with a hormone mixture, which included isobutylmethylxanthine, dexamethasone, and insulin (MDI). The methanol extract of C. crispus significantly decreased fat accumulation by inhibiting adipogenic signal transcriptional factors in MDI-induced 3T3-L1 cells in a dose-dependent manner. In MTT assays and on PI-staining, methanol extract of C. crispus inhibited the proliferation of 3T3-L1 cells during mitotic clonal expansion (MCE). The anti-adipogenic effect of the Carduus extract seemed to be associated with the upregulation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways within the first 2 days after MDI treatment. These results suggest that methanol extract of C. crispus might be beneficial for the treatment of obesity.

• 한국가축번식학회지
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• 제26권4호
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• pp.321-328
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• 2002

The hormone resistin is associated with type II diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downreguates a resistin. Resistin gene expression is induced during adipocyte differentiation and resistin polypeptide is secreted by adipocytes. But, the correlation between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin CDNA and to generate transgenic mice overexpressing mouse resistin gene. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$^{TM}$-mus/resistin by digestion of Bgl II, and has used for microin- jection into pronuclei of one cell embryos. Mouse resistin expression was detected in transgenic F$_1$mice by RT-PCR. The transgenic mouse with resistin gene expression has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, in fasting group, resistin expression was higher than that of re-feeding group. This result demonstrates that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes caused by insulin resistance of resistin.n.