• 동의생리병리학회지
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• 제32권6호
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• pp.384-395
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• 2018

The aim of this study was to investigate whether a novel formulation of an herbal extracts has an inhibitory effect on obesity. To determine its anti-obesity effects, we performed anti-obesity-related experiments in vitro and in vivo. Thus, our present study was carried out to evaluate the anti-obesity effect of herbal extracts using a high fat diet (HFD)-induced obese mouse model and 3T3-L1 adipose cells. The effects of each herbal extracts on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. Results showed that treatment with each herbal extracts at $10{\sim}100{\mu}g/ml$ had no effect on cell morphology and viability. Without evidence of toxicity, herbal extracts treatment decreased lipid accumulation compared with the untreated adipocytes controls as shown by the lower absorbance of Oil Red O stain. Futhermore, compared with control-differentiated mature adipocytes, each herbal extracts significantly inhibited lipid accumulation in mature 3T3-L1 adipocytes. In the HFD-fed obese mice, body weight, liver weight and white adipose tissue weights were significantly reduced by mixture of herbal extracts administration in mouse skin. Futhermore, we found that mixture of herbal extracts administration suppressed serum triglyceride (TG), and total cholesterol (TCHO) in HFD-induced obese mouse model. The mixture of herbal extracts of permeability was estimated by measuring the transepithelial electrical resistance (TEER) value in pig skin. The optimized formulations of herbal extracts (Test 3 formulation) showed skin permeation. However, test 1 formulation containing essential oil as enhancer showed maximum skin permeation. After confirming the enhanced skin permeability, in vivo studies were performed to assess whether skin irritation potential on the basis of a primary irritation index (PII) in rabbit skin. Reactions were scored for erythema/edema reactions at 24 h, 48 h and 72 h post-application. It was concluded that the test 1 formulation was not irritation (PII = 0). The present study suggests that the test 1 formulation might be of therapeutic interest with respect to the treatment of obesity.

• 대한한방소아과학회지
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• 제27권4호
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• pp.17-30
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• 2013

Objectives This experimental study was designed to investigate the effects of Mulberry leaves contained herbal mixture (MLHM) on body weight, serum lipid level and adipocyte differentiation in high fat diet-fed obese mice. Methods Four-week old mice (wild-type C57/BL6) were used for all experiments. Cells were incubated with MLHM at the indicated concentration (0.04-4mg/ml) for 24h, and growth rate was assessed by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. 3T3-L1 preadipocytes were incubated in DMEM for 2 days with the indicated concentrations of MLHM, and on Day 6, the cells were fixed and the cellular lipid contents were assessed by Oil-Red-O staining. The expression of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR ${\gamma}$) and cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$) as adipocyte-specific proteins were determined by real time RT-PCR and western blotting. In addition, body weight gain and serum lipid levels were measured in the mice with obesity induced by the high fat-diet for four weeks. Results Though MLHM did not show toxicity even at the concentration of 4mg/ml, MLHM significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner. Also, MLHM significantly reduced the expressions of PPAR ${\gamma}$ and C/EBP ${\alpha}$ in a dose-dependent manner. Furthermore, MLHM significantly reduced body weight gain and LDL-cholesterol contents in high fat diet-fed obese mice. Conclusions These results demonstrate that MLHM exerts anti-obesity effect in 3T3-L1 cells and mice with obesity by high-fat diet.

• 대한의용생체공학회:의공학회지
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• 제41권3호
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• pp.138-145
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• 2020

Adipocytes are the main constituent of adipose tissue. Understanding the molecular basis of adipogenesis is pivotal to finding the therapeutic targets for treatment of obesity. Melatonin is associated with obesity and its mechanism is currently under intensive investigation. The objective of this study was to investigate the effect of melatonin on adipogenesis in differentiating preadipocytes. 3T3-L1 preadipocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 5% calf serum at 37℃ with 5% CO₂ in a humidified incubator. Differentiation was induced using DMEM with 10% fetal bovine serum supplemented with MDI two days after cell confluence (day 0). Cells were treated with 0, 10 and 100 μM melatonin on either day 0 or day 5. 72 hours after each treatment, lipid accumulation was measured by oil red O staining. Proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to membranes. As a result, lipid accumulation decreased with melatonin treatment. ERK pathway, activated when differentiation is induced, also decreased with an increase in melatonin concentration. Furthermore, the expression of key adipogenic factors, C/EBPα, C/EBPβ, and PPARγ, were reduced by melatonin treatment. These results imply that melatonin may inhibit the process of adipogenesis and may have a role as a new anti-obesity agent.

• 한방비만학회지
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• 제19권2호
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• pp.106-112
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• 2019

Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4^th day and insulin medium at 6^th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×10⁷ CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

• Nutrition Research and Practice
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• 제8권1호
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• pp.33-39
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• 2014

Obesity occurs when a person's calorie intake exceeds the amount of energy burns, which may lead to pathologic growth of adipocytes and the accumulation of fat in the tissues. In this study, the effect and mechanism of pear pomace extracts on 3T3-L1 adipocyte differentiation and apoptosis of mature adipocytes were investigated. The effects of pear pomace extract on cell viability and the anti-adipogenic and proapoptotic effects were investigated via MTT assay, Oil red O staining, western blot analysis and apoptosis assay. 3T3-L1 preadipocytes were stimulated with DMEM containing 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), $5{\mu}g/ml$ insulin and $1{\mu}M$ dexamethasone for differentiation to adipocytes. 3T3-L1 cells were cultured with PBS or water extract of pear pomace. Water extract of pear pomace effectively inhibited lipid accumulations and expressions of PPAR-${\gamma}$ and $C/EBP{\alpha}$ in 3T3-L1 cells. It also increased expression of p-AMPK and decreased the expression of SREBP-1c and FAS in 3T3-L1 cells. The induction of apoptosis was observed in 3T3-L1 cells treated with pear pomace. These results indicate that pear pomace water extract inhibits adipogenesis and induces apoptosis of adipocytes and thus can be used as a potential therapeutic substance as part of prevention or treatment strategy for obesity.

• Toxicological Research
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• 제23권3호
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• pp.223-229
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• 2007

Conjugated linoleic acid (CLA) has been recently reported to have an anti-obesity effect in animals and humans. The objective of this study was to investigate effects of diglyceride (DG)-CLA on proliferation and differentiation of 3T3-L1 preadipocytes. Cell proliferation was determined using WST-8 analysis and cell differentiation was determined by glycerol-3-phosphate dehydrogenase (GPDH) activity. Lipid accumulation in differentiating 3T3-L1 cells was determined by Oil red O staining. There were four experimental groups including vehicle control (DMSO), CLA, triglyceride (TG)-CLA, and DG-CLA. Treatments of CLA, TG-CLA, and DG-CLA at the concentrations of $10{\sim}1000{\mu}g/ml$ reduced proliferation of preconfluent 3T3-L1 cells in a dose-dependent manner. Among them CLA was the most effective in the proliferation inhibition of preconfluent 3T3-L1 cells with increasing concentrations. Treatments of CLA and DG-CLA at the concentration of $100{\mu}g/ml$ significantly inhibited differentiation of postconfluent 3T3-L1 cells as measured by GPOH activity (p<0.05). In addition, treatments of CLA, TG-CLA, and DG-CLA effectively inhibited lipid accumulation during differentiation of 3T3-L 1 cells. OG-CLA had the most inhibitory effect on the differentiation and lipid accumulation. These results suggest that the compounds including CLA have a respectable anti-obesity effect and that consumption of DG-CLA as a dietary oil may give a benefit for controlling overweight in humans.

• 대한의용생체공학회:의공학회지
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• 제45권1호
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• pp.43-48
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• 2024

The prevalence of obesity and its complications is steadily increasing worldwide. It is essential to understand cellular level metabolism and microenvironment to treat diseases related to lipid metabolism. Mechanical loading can activate signaling pathway by stimulating cells, especially vibration loading known to inhibit adipogenesis, so it has been studied as a treatment for obesity. Also, vibration loading can affect the inside of the human body non-invasively. Another clue to reducing adipose tissue is browning, which means that white adipocytes changes to brown adipocyte. In this study, we design and developed a device that that can control cell-cell contact, and vibration simulation device. Using these two devices, we investigated responses of cells to vibration loading. Protein expression associated with browning and adipogenesis were analyzed. In conclusion, vibration loading can be transmitted through cell contact and loading applied to the cells can induce browning and inhibit adipogenesis of preadipocytes. These results suggest the possibility that vibrations could be a treatment for obesity.

• International Journal of Industrial Entomology and Biomaterials
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• 제21권2호
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• pp.169-174
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• 2010

Hydrolysate silk fibroin (HSF) is a fibrous protein composed of parallel $\beta$-structures and is made from pure silk elements including 18 amino acids, with glycine, alanine, and serine comprising of over 80% of the amino acids. Numerous studies have documented a range of effects of HSF, including moisturizing, antioxidant activity, nervous system disorders, and many more. We investigated whether HSF has anti-obesity effects in vitro. The effects of HSF inhibition on lipid accumulation and acceleration of lipid degradation in 3T3-L1 cells were studied. Treatment of 3T3-L1 cells with HSF caused significant inhibition of cell viability, an increase in glycerol release, and a decreased in adipocyte differentiation. Moreover HSF stimulated downregulated of adipogenic enzyme expressions (PPAR${\gamma}$ and C/EBP${\alpha}$) and up-regulated of fatty oxidation enzyme expressions (CPT-1 and UCP-2). Based on these results, hydrolysate silk fibroin can be suggested as a potential therapeutic substance as part of a prevention or treatment strategy for obesity.

• 생명과학회지
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• 제30권6호
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• pp.532-541
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• 2020

본 연구는 지방감소를 위한 소재개발로 감국 유산균 발효물이 갖는 지방구세포 분화 억제효과를 관찰하였다. 감국 추출물의 세포독성을 극복하는 유산균의 발효물을 제작하였다. 3T3-L1 세포주에서 감국 추출물 및 발효물이 갖는 세포독성은 모두 없었다(1day culture). 감국 추출물 처리 대조군과 비교하여 3T3-L1 세포주에 처리시 증식 유도된 발효물을 선별하였다. 감국 추출물 및 발효물의 분화억제 및 세포생존률 FACS분석은 분화 유도된 세포가 모든 실험군에서 줄어들었다. 3T3-L1 세포주에서 감국 추출물과 발효물 처리가 protein kinase B (Akt) pathway활성이 증가하였고, 단백질 발현은 지방구세포로 분화되면서 Gli2의 수준은 감소하였다. Hedgehog를 조절하는 유산균은 KCTC 3115인 것을 알 수 있었다. 분화와 관련된 KCTC 3115 및 KCTC 3109 발효군에서 단백질 수준에서 C/EBPα 및 FAS를 감소, pACC는 증가시키는 것을 확인하였다. 감국 추출물과 4개의 감국 유산균 발효물 중 Lactococcus lactis subsp. lactis KCTC 3115 발효물이 지방구세포 분화 신호를 더 효과적으로 조절하고, hedgehog을 같이 조절하여 지방전구세포의 분화를 억제하는 것을 알 수 있었다. Hedgehog 신호를 조절하면서 분화를 억제하는 물질에 대한 연구가 더 필요할 것으로 판단된다. 따라서 감국 발효물의 생리활성 물질 중 향후 매커니즘 분석을 위한 활성물질의 자료가 더 필요할 것으로 여겨지며, 감국 추출물 및 감국 발효물의 hedgehog 신호조절이 새로운 비만치료제로 개발될 수 있음을 위한 가능성을 제시하고자 한다.

• Journal of Nutrition and Health
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• 제39권8호
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• pp.748-755
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• 2006

The purpose of this study was to find out effects of treatment of ginsenoside Re, Rc and EGCG on mRNA expressions of leptin, hormone sensitive lipase (HSL) and resistin in 3T3-L1 adipocytes. The concentrations of EGCG were treated with $0.01{\times}10^{-7},\;0.1{\times}10^{-7},\;1{\times}10^{-7}\;and\;1{\times}10^{-6}\;or\;100{\mu}g/ml$ ginsenoside Re, Rc in culture cell for 13 days. mRNA expression of leptin wasn't expressed in preadipocyte but according to differentiation of adipocyte, the that of mRNA expression was decreased at gensenosids or EGCG treated cells compared with non treated adipocyte. Expression of HSL mRNA was increased in G-Re, G-Rc and EGCG treated cells compared with non treated cells. The resistin level was significantly decreased in adipocytes treated with G-Re, G-Rc and EGCG. These pattern was similar to leptin expression. These results support that treatment of gensenosides or EGCG in 3T3-L1 adipocyte resulted to affect of leptin and resistin as well as HSL mRNA levels, accordingly, levels of leptin and HSL will be acted by signalling body fat stores to the hypothalamus which in turn regulates food intake andenergy expenditure to maintain body weight homeostasis. And also regulation of resistin mRNA will prevent to diabetics attacked with obesity. In conclusion, we suggest that consumption of ginseng saponine or EGCG might prevent human diabetics or/and obesity.