• Archives of Plastic Surgery
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• 제32권4호
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• pp.533-538
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• 2005

Due to its safety and softness, autologous fat transplantation has been commonly performed for soft tissue correction. However, the injected fat is absorbed resulting in the reduction of volume of the graft by 40-60% within a few months. Thus, there was an attempt to use adipocytes differentiated from preadipocytes in vitro for transplantation. Differentiated adipocytes were biocompatible and matured with gradual volume increase at transplantation site in clinical study(unpublished data). In addition, they did not induce immune rejection in response to nonself lymphocytes in a mixed lymphocyte reaction(MLR)(unpublished data). The purpose of this study is to differentiate mouse preadipocytes following labeling into adipocytes to establish an animal model for allogenic transplantation. Preadipocytes isolated from inguinal and retroperitoneal fat pad of C57BL/6 mice were proliferated with growth medium by passage 3 and differentiated into adipocytes with different culture conditions after labeled with BrdU. At most suitable conditions, above 90% of preadipocytes were differentiated and BrdU labeling did not affect differentiation rate and function of differentiated adipocytes. These results demonstrate that BrdU-labeled adipocytes resulting from this in vitro differentiation protocol are useful for allogenic transplantation study.

• Asian-Australasian Journal of Animal Sciences
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• 제13권4호
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• pp.446-450
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• 2000

The experiment was conducted to study the effects of water soluble vitamins and retinoic acid on the differentiation of preadipocyte from omental, subcutaneous, intermuscular and intramuscular adipose tissue of Hanwoo. Differentiation was assessed by the change in enzyme activity, glycerol-3 phosphate dehydrogenase in serum free cell culture system. Preadipocytes treated with biotin ($10{\mu}M$) and pantothenic acid ($100{\mu}M$) were significantly (p<0.05) less differentiated than those from the control in all adipose tissue depots except intramuscular tissue. Although there was no significance, vitamin C was shown to stimulate the adipocyte conversion in omental and subcutaneous, but not in intermuscular and intramuscular adipose tissues. Lower values of GPDH activity in intermuscular preadipocyte were interpreted to be caused by relatively higher amounts of protein. In this experiment vitamin C did not stimulate fat deposition in intramuscular adipose tissue but further experiments are needed on the role of vitamin C in preadipocyte differentiation. When treated with different levels of retinoic acids, differentiation of preadipocytes was significantly (p<0.05) reduced from the level of $0.5{\mu}g/ml$ in omental and intermuscular, from $50{\mu}g/ml$ in subcutaneous, and in intramuscular at $500{\mu}g/ml$, thus showing that intramuscular preadipocytes were least responsive to retinoic acid in differentiation. All-trans retinoic acid appeared to inhibit the differentiation in a dose dependent manner, regardless of adipose tissues type.

• 동의생리병리학회지
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• 제23권6호
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• pp.1409-1415
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• 2009

Obesity is especially a serious health problem in industrialized countries, because it is considered to be a risk factor associated with the genesis or development of various metabolic diseases, including cardiovascular disease and type 2 diabetes mellitus. The purpose of this study was to investigate the effects of tanshinone IIA from Salvia miltiorrhiza Bunge on induction of apoptossis and inhibition of adipogenesis in in 3T3-L1 preadipocytes and adipocytes. The results demonstrated that tanshinone IIA decreased cell population growth of 3T3-L1 preadipocytes, assessed with the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and LDH (lactate dehydrogenase) assay. Flow cytometric analysis of 3T3-L1 preadipocytes exposed to tanshinone IIA showed that apoptotic cells increased in a timeand dose-dependent manner. Treatment with tanshinone IIA decreased the number of normal cells and increased the number of apoptotic cells in a dose-dependent manner. The induction of apoptosis in 3T3-L1 preadipocytes by tanshinone IIA was mediated through the activation of caspase-3 and Bax, and then through the cleavage of PARP and the down-regulation of Bcl-2. Moreover, tanshinone IIA significantly decreased the amount of intracellular triglycerides and GPDH (glycerol-3-phosphate dehydrogenase) activity in 3T3-L1 adipocytes. Our results suggest that tanshinone IIA efficiently induces apoptosis and inhibits adipogenesis in 3T3-L1 preadipocytes and adipocytes.

• 한방비만학회지
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• 제23권2호
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• pp.69-77
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• 2023

Objectives: Atriplex gmelinii C. A. Meyer is a halophyte belonging to the Chenopodiaceae family, and its young leaves and stems are used as fodder for livestock. The aim of the present study was to investigate the effects of A. gmelinii extract and its solvent fractions on lipid accumulation during adipogenesis of 3T3-L1 preadipocytes. Methods: The samples of A. gmelinii were separately extracted using methylene chloride and methanol. Subsequently, they were combined to formulate the initial extract, which was then partitioned based on polarity to prepare solvent fractions. Oil Red O staining was employed to measure lipid accumulation during the differentiation of 3T3-L1 preadipocytes. To verify cytotoxicity in 3T3-L1 cells, MTT assays were conducted. The expression levels of transcription factors in 3T3-L1 preadipocytes were measured through Western blotting analysis. Results: At 50 ㎍/mL, treatment of A. gmelinii extract and its solvent fractions during the differentiation of 3T3-L1 preadipocytes significantly diminished lipid accumulation with no noteworthy cytotoxicity on cell viability. Additionally, when investigating the biochemical pathways that underlie the prevention of lipid accumulation using solvent fractions, it was found that the n-BuOH and n-hexane fractions significantly decreased the expression of key transcription factors involved in the generation of fat, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), and sterol regulatory element-binding protein-1c (SREBP1c). Conclusions: These findings indicate that A. gmelinii can effectively reduce the accumulation of fat in 3T3-L1 adipocytes, making it a potentially valuable material for mitigating and preventing obesity.

• IMMUNE NETWORK
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• 제16권3호
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• pp.189-194
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• 2016

Obesity is characterized as an accumulation of adipose tissue mass represented by chronic, low-grade inflammation. Obesity-derived inflammation involves chemokines as important regulators contributing to the pathophysiology of obesity-related diseases such as cardiovascular disease, diabetes and some cancers. The obesity-driven chemokine network is poorly understood. Here, we identified the profiles of chemokine signature between human preadipocytes and adipocytes, using PCR arrays and qRT-PCR. Both preadipocytes and adipocytes showed absent or low levels in chemokine receptors in spite of some changes. On the other hand, the chemokine levels of CCL2, CCL7-8, CCL11, CXCL1-3, CXCL6 and CXCL10-11 were dominantly expressed in preadipocytes compared to adipocytes. Interestingly, CXCL14 was the most dominant chemokine expressed in adipocytes compared to preadipocytes. Moreover, there is significantly higher protein level of CXCL14 in conditioned media from adipocytes. In addition, we analyzed the data of the chemokine signatures in adipocytes obtained from healthy lean and obese postmenopausal women based on Gene Expression Omnibus (GEO) dataset. Adipocytes from obese individuals had significantly higher levels in chemokine signature as follows: CCL2, CCL13, CCL18-19, CCL23, CCL26, CXCL1, CXCL3 and CXCL14, as compared to those from lean ones. Also, among the chemokine networks, CXCL14 appeared to be the highest levels in adipocytes from both lean and obese women. Taken together, these results identify CXCL14 as an important chemokine induced during adipogenesis, requiring further research elucidating its potential therapeutic benefits in obesity.

• Journal of Animal Science and Technology
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• 제50권5호
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• pp.649-656
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• 2008

본 연구는 insulin-like growth factors(IGFs)가 돼지 지방전구세포의 증식과 분화에 미치는 작용기전을 구명하기 위해서 수행하였다. 지방전구세포는 갓난 암퇘지의 등지방에서 분리하였고, serum-deprived 조건하에서 IGFs와 mutant IGFs를 함유시켜 배양했는데 이 mutant IGFs는 IGF-Ⅰ에 비해 type-1 IGF receptor와 insulin receptor에 대한 친화력이 낮다. 50ng/ml의 IGF-Ⅰ, [Leu60]IGF-I, IGF-Ⅱ 및 [Leu27]IGF-Ⅱ를 배양중인 세포에 4일동안 처리했다. IGF-Ⅰ, [Leu60]IGF-I, IGF-Ⅱ 및 [Leu27]IGF-Ⅱ는 돼지 지방전구세포의 증식을 각각 39%, 8%, 25% 및 2% 촉진했다(증가된 세포수에 의해 측정). 이 사실은 IGF-Ⅰ과 IGF-Ⅱ는 type-1 IGF receptor 또는 insulin receptor에 결합을 통해서 지방세포의 증식 촉진을 가져왔음을 나타낸다. 그리고 IGF-Ⅰ, [Leu60]IGF-I, IGF-Ⅱ 및 [Leu27] IGF-Ⅱ는 지방전구세포의 분화를 50%, 17%, 37% 및 30% 각각 촉진시켰다(세포 분화는 glycerol- phosphate dehydrogenase 활성도에 의해 측정했다). IGF-Ⅰ의 type-1 IGF receptor 또는 insulin receptor에의 친화력이 낮아져서 세포 분화 촉진작용을 감소시킨 것이다. 그러나 [Leu27] IGF-Ⅱ의 분화촉진 작용은 IGF-Ⅱ의 그것에 비해 크게 차이가 나지 않았는데, 이 사실은 IGF-Ⅰ과 IGF-Ⅱ는 서로 다른 수용체－매개 작용기전에 의해 세포분화를 촉진시킴을 나타낸다. 즉 IGF-Ⅱ는 type-1 IGF receptor 또는 insulin receptor가 관여하지 않는 작용을 통해 돼지 지방전구세포의 분화를 촉진시켰다. 이 작용은 IGF-Ⅱ가 type-2 IGF receptor(또는 cation- independent mannose-6 phosphate receptor [CIM6P /IGF2 receptor])에 결합을 통해서 이뤄지는 것으로 여겨진다. 위의 결과는 IGF-Ⅱ가 CIM6P/ IGF2 receptor에의 결합을 통해 동물 지방전구세포의 분화를 촉진시킨다는 것을 밝혀낸 최초의 연구이다. 요약하면 이 본 연구는 IGF-Ⅰ과 IGF-Ⅱ는 서로 다른 세포내 receptor가 관여하는 작용기전을 통해 돼지 지방전구세포의 분화를 촉진함을 보여준다.

• 대한약침학회지
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• 제10권3호
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• pp.29-35
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• 2007

Objectives The purpose of this study is to investigate the effects of cultivated wild ginseng extract on primary cultured preadipocyte and adipocytes. Methods Diminish preadipocyte proliferation does primary role to reduce obesity. So, preadipocytes and adipocytes were performed on cell cultures with using Sprague-Dawley rats and treated with 0.01-1mg/ml cultivated wild ginseng extract. Result At all concentrations, cultivated wild ginseng extract wasn't show the suppress proliferation of preadipocytes significantly and failed to show effects on decomposition of adipocytes except high dosage. Conclusion Based on these findings, cultivated wild ginseng is not a suitable choice for the treatment of localized obesity.

• 생명과학회지
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• 제27권10호
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• pp.1104-1110
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• 2017

히스톤 H3K4의 메칠화는 3T3-L1의 지방세포의 분화를 촉진하는 것으로 알려져 있으나, 히스톤 H3K4 메칠화 효소인 SET1A가 지방세포 분화를 조절하는지에 대해서는 보고된 바가 없다. 그러므로 본 연구에서는 SET1A의 3T3-L1 지방세포의 분화조절과 기전을 연구하였다. SET1A의 발현은 3T3-L1 지방세포 분화과정에서 증가함을 관찰하였다. 3T3-L1 지방전구세포에서 siRNA을 이용하여 SET1A의 발현을 감소시키면 3T3-L1 지방전구세포의 분화가 억제됨을 관찰하여 SET1A가 3T3-L1 지방전구세포의 분화를 촉진함을 알 수 있었다. 이에 대한 조절기전을 알기 위해, SET1A의 발현을 감소시킨 3T3-L1 지방전구세포의 세포증식을 측정한 결과, 분화 초기 단계인 분화 후 2일 동안 3T3-L1 지방세포의 증식이 감소하였다. 또한 분화 후 7일 동안 지방세포세포 분화 조절인자들의 발현을 측정한 결과, SET1A의 발현을 감소시킨 3T3-L1 지방세포에서 $PPAR{\gamma}$의 발현이 감소하였다. 위와 같은 연구결과를 바탕으로, SET1A는 분화초기단계에서는 mitotic clonal expansion 단계를 촉진하고, 분화후기단계에서는 $PPAR{\gamma}$의 발현을 증가시켜 3T3-L1 지방세포의 분화를 촉진함을 알 수 있었다.

• 생명과학회지
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• 제18권4호
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• pp.537-541
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• 2008

백색지방조직(white adipose tissue)은 에너지 저장뿐만 아니라 다양한 adipokines을 분비하는 중요한 내분비 기관이다. 급성기반응 단백질로 알려져 있는 합토글로빈(haptoglobin)도 adipokine의 한 종류로서 지방세포에서 합성되고 분비된다. 그러나 adipokine으로서의 기능과 지방조직에서의 역할은 아직까지 규명되지 않았다. 본 연구에서는 3T3-L1 지방전구세포를 합토글로빈 유전자로 transfection 시켜 합토글로빈을 과발현하는 세포를 만들고 세포증식, 염증관련 인자들의 발현조절 및 단구세포의 유인성을 조사하였다. 그 결과, 합토글로빈은 3T3-L1 세포의 성장에는 별 영향을 미치지 않았으나 IL-6와 COX-2 발현을 저해하고 HO-1 합성을 증가하였다. 또한 THP-1 단구세포를 이용한 invasion assay에서는 합토글로빈이 단구세포의 이동을 저해하였다. 이러한 결과들은 합토글로빈이 지방조직에서 항염증 반응에 관여함을 시사한다. 만성적 염증상태(chronic low-grade inflammatory state)로 인식되고 있는 비만은 염증관련 인자들에 의한 인슐린저항성이 유도되는 바, 합토글로빈은 비만 관련 인슐린저항성을 방지하고 인슐린민감성을 향상시키는 데에도 기여할 것으로 생각된다.

• Nutritional Sciences
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• 제6권3호
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• pp.148-154
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• 2003

Although various raw plant materials have been demonstrated to exert anti-obesity effects to a greater or lesser extent in both humans and animals when they are used to supplement the diet, it has not been shown extensively that they influence adipocyte cell differentiation involving lipid metabolic gene expressions. Using a well-established 3T3-L1 preadipocyte differentiation system, we decided to look into molecular and cellular event occurring during adipocyte differentiation when raw plant materials aye included in the process, in an effort to demonstrate the potential use of a screening system to define the functions of traditionally well-known materials. To these ends, the effects of ethanol (EtOH) or EtOH/distilled water (DW) extracts of Wax Gourd were examined using cytochemical and molecular analyses to determine whether components of the extracts modulate adipocyte differentiation of 3T3-Ll preadipocytes in vitro. The cytochemical results demonstrated that EtOH or EtOH/DW extracts did not affect lipid accumulation and cell proliferation, although the degree of lipid accumulation was influenced slightly depending on the extract. EtOH extract was highly effective in apoptotic induction during differentiation of 3T3-Ll preadipocytes (p<0.05). Reverse transcription-polymerase chain reaction (RT-PCR) analysis of lipoprotein lipase (LPL), Uncoupling protein (Ucp) 2, 3 and 4 also showed that while LPL expression was not influenced, Ucp2, 3 and 4 were up regulated in the EtOH extract-treated group and down regulated in the EtOH/DW extract-treated group. These changes in gene expressions suggest that the components in different fractions of Wax Gourd extracts may modulate lipid metabolism by either direct or indirect action. Taking these results together, it was concluded that molecular and cellular analyses of adipocyte differentiation involving lipid metabolic genes should facilitate understanding of cellular events occurring during adipocyte differentiation. Furthermore, the experimental scheme and analytical methods used in this study should provide a screening system for the functional study of raw plant materials in obesity research.