• 한국지역사회생활과학회지
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• 제27권4호
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• pp.863-873
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• 2016

The current study was undertaken to determine the effects of the ethanol extracts of loquat (Eriobotrya japonica Lindl.) seeds, flesh or leaves on the differentiation of 3T3-L1 cells and male pig preadipocytes. The cell number was measured with the MTT assay after trypsin digestion. The cell differentiation was determined by measuring the glycerol-3-phosphate dehydrogenase (GPDH) activity and triglyceride(TG) content. No cytotoxicity was observed from the loquat flesh and leaf ethanol extracts at concentrations of 5, 10, 25, 50, 100 or $200{\mu}g/mL$ in 3T3-L1 cells and pig preadipocytes. However, the cell viability of neither cell line were affected by up $50{\mu}g/mL$ of loquat seed ethanol extract. Treatment with the loquat seed and leaf ethanol extracts significantly suppressed the terminal differentiation of both cell lines in a dose-dependent manner, as confirmed by the decrease in the glycerol-3-phosphate dehydrogenase(GPDH) activity and TG content. Treatment with the loquat seed and leaf ethanol extracts inhibited the GPDH activity and reduced the TG content of both cell types more effectively than that with the loquat flesh ethanol extract. The most potent anti-adipogenic effect was obtained in the case of the ethanol extract of loquat seeds.

• Toxicological Research
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• 제31권2호
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• pp.191-201
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• 2015

We evaluated the inhibitory effect of Pueraria lobata root ethanol extract (PLREE) on lipid accumulation during 3T3-L1 differentiation to adipocytes by measuring the intracellular expression of adipogenic, lipogenic, and lipolytic markers and lipid accumulation. The total polyphenol and flavonoid content of PLREE were 47 and 29 mg/g, respectively. The electron donating capacity of PLREE at $1,000{\mu}g/mL$ was 48.8%. Treatment of 3T3-L1 preadipocytes with 100, 250, or $500{\mu}g/mL$ PLREE for 8 days dose-dependently promoted the differentiation of 3T3-L1 cells. In contrast, the lipid content of PLREE-treated cells was significantly reduced by 7.8% (p < 0.05), 35.6% (p < 0.001), and 42.2% (p < 0.001) following treatment with 100, 250, and $500{\mu}g/mL$ PLREE, respectively, as compared to differentiated control cells. PLREE upregulated peroxisome proliferator-activated receptor ${\gamma}$ mRNA and protein, and sterol regulator element-binding protein-1c mRNA levels, but did not affect CCAAT/enhancer binding-protein ${\beta}$ and ${\alpha}$ mRNA levels. PLREE also downregulated acetyl-CoA carboxylase mRNA and protein, fatty acid synthase (FAS) protein, and leptin mRNA levels, but did not affect FAS mRNA expression. PLREE upregulated adipose triglyceride lipase mRNA and protein expression, and hormone-sensitive lipase (HSL) protein expression, but did not affect HSL mRNA expression. In conclusion, we found that PLREE enhanced adipogenesis, but reduced lipogenesis, resulting in decreased lipid accumulation in 3T3-L1 cells.

• 한방비만학회지
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• 제20권2호
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• pp.78-87
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• 2020

Objectives: To expand and provide information on the efficacy of herbal medicines, anti-obesity effects were evaluated. In many studies, plant-derived components with anti-obesity efficacies have been investigated for their potential inhibitory effects on 3T3-L1 preadipocyte cells. The purpose of this study was to investigate the anti-obesity effects of 65 herbal medicine in 3T3-L1 preadipocyte cells. Methods: Preferentially, 3T3-L1 cells were treated with 65 herbal medicines (500 ㎍/mL) during differentiation for 8 days. Next, 3T3-L1 cells were treated with selected herbal medicines at concentrations ranging from 50 to 200 ㎍/mL during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining. The expressions of genes related to adipogenesis were measured by reverse transcription polymerase chain reaction and Western blot analyses. Results: Among the 65 kinds of herbal medicines, 13 herbal medicines that been shown to be effective against the accumulation of lipid droplets were selected. Finally, selected Banhasasim-tang and Samhwangsasim-tang showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affecting cell toxicity. In addition, Banhasasim-tang and Samhwangsasim-tang significantly reduced the expression levels of several adipocyte marker genes including peroxisome proliferator activated receptor-γ and CCAAT/enhancer binding protein-α. Conclusions : These results suggest that the ability of Banhasasim-tang and Samhwangsasimtang has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. Banhasasim-tang and Samhwangsasim-tang may be a promising medicine for the treatment of obesity and related metabolic disorders.

• 동의생리병리학회지
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• 제18권2호
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• pp.451-456
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• 2004

Based on the data from our previous studies, four new diabetic remedies were composed with the addition of Coicis Semen into Okchun-san (OCH), Commelinae Herba into Gangsim-tang (GST) , Scrophulariae Radix into Sunki-san (SKS), and Erythrinae Cortex into Yukmijihuang-hwan (YMG). The water extracts of these new remedies were treated in 3T3-L1 fibroblasts and adipocytes in order to investigate insulin-like substances and insulin sensitizers, respectively. With and without differentiation inducers, unmodified SKS (SKS-O) treatment induced 3T3-L1 fibroblasts into adipocytes more than the control. However, without inducers, YMG treatment, but not SKS, induced the differentiation more than the control among modified remedies. Without inducers, SKS, OCH as well as YMG increased the induction of differentiation from 3T3-L1 fibroblasts into adipocytes, compared to the control. The treatment of OCH and YMG with 1 ng/mL insulin increased glucose uptake much more than only insulin 1 ng/mL treatment. Thus, OCH and YMG contained increased insulin actions. In conclusions, the modified remedies, OCH and YMG, contained insulin-like substances and insulin sensitizers, and they can be improved the hypoglycemic effects.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1327-1331
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• 2008

In order to determine whether peach contains compounds to regulate adipocyte differentiation, extracts of flesh/pericarp of yellow/white peach were prepared in water, ethyl acetate (EtOAc), or n-butanol solvent and determined for effects on adipocyte differentiation in C3H10T1/2 or 3T3-L1 cells. Interestingly, none of peach extracts has statistically significant stimulatory effect on the adipocyte differentiation in C3H10T1/2. Furthermore, the presence of EtOAc extract of white peach pericarp (WPP) was found to inhibit lipid accumulation in 3T3-L1 cells both by microscopic examination of Oil Red O-stained lipid droplets and by spectrophotometric quantification of extracted stain, indicating a significant inhibitory effect on adipocyte differentiation. The inhibition of lipid accumulation was accompanied by a significant decrease in the expression levels of adipocyte molecular markers-peroxisome proliferator-activated receptor $\gamma$, CAAT enhancer binding protein $\alpha$, and fatty acid-binding protein. Thus, this study determined that WPP EtOAc extract contains the inhibitory compound(s) on adipogenesis.

• 동의생리병리학회지
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• 제24권1호
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• pp.118-123
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• 2010

Piper nigrum Linne has been used spice as well as herbal medicine in worldwide and has function of anti-oxidant, anti-inflammation, anti-cancer, bioavailability of drugs and etc. In this study, anti-adipogenic activity of Piper nigrum Linne extract and its constituent piperine were investigated in 3T3-L1 differentiation. Adipogenic effects and lipid accumulation in 3T3-L1 was measured by RT-PCR and Oil Red O staining assays respectively in stimulation of Piper nigrum Linne extract and piperine. Piper nigrum Linne and piperine reduced lipid accumulation in 3T3-L1 differentiation and expression of genes associated with adipogenesis such as PPAR${\gamma}$, adipsin, SERBP-1c and LPL. All of taken, anti-adipogenic mechanism of Piper nigrum Linne and piperine were related with regulation of SREBP-1c and PPAR${\gamma}$ expression.

• Journal of Microbiology and Biotechnology
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• 제18권1호
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• pp.80-87
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• 2008

The 3T3-L1 cell line is a well-established and commonly used in vitro model to assess adipocyte differentiation. Over the course of several days, confluent 3T3-L1 cells can be converted to adipocytes in the presence of an adipogenic cocktail. In this study, the effects of chitosan oligosaccharides (CO) on adipocyte differentiation of 3T3-L1 cells were studied. The CO significantly decreased lipid accumulation, a marker of adipogenesis, in a dose-dependent manner. The low molecular mass CO (1-3 kDa) were the most effective at inhibiting adipocyte differentiation. Moreover, mRNA expression levels of both CCAAT/enhancer-binding protein (C/EBP) ${\alpha}$ and peroxisome proliferator-activated receptor (PPAR) ${\gamma}$, the key adipogenic transcription factors, were markedly decreased by CO treatments. CO also significantly down regulated adipogenic marker proteins such as leptin, adiponectin, and resistin. Our results suggest a role for CO as antiobesity agents by inhibiting adipocyte differentiation mediated through the down regulated expression of adipogenic transcription factors and other specific genes.

• Asian-Australasian Journal of Animal Sciences
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• 제29권3호
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• pp.419-427
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• 2016

There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at $42^{\circ}C$ for one hour and then allowed to recover at normal incubation temperature of $37^{\circ}C$ for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to $400{\mu}g/mL$) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

• 대한한방소아과학회지
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• 제27권4호
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• pp.108-121
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• 2013

Objective This study was designed to investigate the effects of Gamiygin-tang (GY) extract (GYE) and fermented solution (GYF) on body weight, serum lipid level and adipocyte differentiation in high fat diet-fed obese mice. Materials and Methods High fat diet-fed obese mice and 3T3-L1 adipocytes mice were treated with GYE and GYF and obesity related markers were assessed. A cytotoxicity assay was carried out by MTS assay. Inhibitory effects of GYE and GYF on adipocyte differentiation were carried out by Oil Red O staining. The effects of GYE and GYF on the expression of adipocyte differentiation regulatory factors, peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer binding protein alpha (CEBP-${\alpha}$) were measured by real-time reverse transcriptase-polymerase chain reaction. The effects of GYE and GYF on the expression of adipocyte differentiation regulatory factors were also determined in relation to protein production/protein levels by western blotting. The anti obesity effects of GYE and GYF were measured in high fat-diet induced obese mice. Various factors were measured from the serum of the high fat-diet mice. Results Though GYE did not show toxicity at the concentration of 1mg/ml, GYF showed toxicity at the concentration of 1mg/ml. The GYE at 0.1 and 1mg/ml inhibited the differentiation of 3T3-L1 cells, and the GYF also inhibited the differentiation of 3T3-L1 cells. The effect of GYE on adipocyte differentiation factors including PPAR-${\gamma}$ and CEBP-${\alpha}$ was investigated and compared to the corresponding concentration levels of GYF. GYE and GYF both suppressed the RNA and protein levels of adipocyte differentiation factors. In the animal test both GYE and GYF reduced weight gain. GYE and GYF reduced blood cholesterol, TG and LDL levels. GYF better reduced blood cholesterol levels. Conclusions These results demonstrate that GYE and GYF exerts anti-obesity effect in 3T3-L1 cells and obese mice induced by high-fat diet.

• 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.