• Journal of Korean Medicine for Obesity Research
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• v.13 no.2
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• pp.74-83
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• 2013

Objectives: This study was performed to evaluate the effects of fermented lotus extracts on the inhibition of differentiation in 3T3-L1 preadipocytes. Methods: Extracts of lotus leaf and lotus root were fermented using 4 different probiotics separately, including Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium breve, and Bifidobacterium longum. Inhibition of preadipocyte differentiation was examined by Oil red O dye staining. Expressions of adipogenic transcription factors including CCAAT/enhancer binding proteins (C/$EBP{\alpha}$) and peroxisome proliferators-activated receptor ${\gamma}$ ($PPAR{\gamma}$) were analyzed by real time polymerase chain reaction and Western blotting analysis. Results: Fermented lotus extracts inhibited adipogenic transcription factors by inhibiting preadipocytes differentiation. All of the groups fermented by 4 kinds of probiotics showed reduction in Oil Red O dye staining. Bifidobacterium breve showed the most effective inhibition of C/$EBP{\alpha}$. Bifidobacterium breve and Bifidobacterium longum showed the best downregulation of $PPAR{\gamma}$ expressions compared with the control and the unfermented lotus group. Conclusions: Fermented lotus extracts showed significant effects on inhibition of preadipocyte differentiation in 3T3-L1 preadipocytes showing correlation with insulin sensitivity and lipid metabolism related with obesity.

• The Korean Journal of Community Living Science
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• v.27 no.4
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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.

• The Journal of Pediatrics of Korean Medicine
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• v.29 no.1
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• pp.1-14
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• 2015

Objectives This experimental study was designed to investigate the effects of Dai-saiko-to (DSH) on differentiation of 3T3-L1 preadipocytes and body weight, serum lipid levels in high-fat diet-induced obese mice. Materials and Methods Cells were incubated with DSH at an indicated concentration (0.01-1 mg/ml) for 24h, then the growth rate was assessed by MTS assay. 3T3-L1 preadipocytes were incubated in DMEM for 2 days with the indicated concentrations of DSH. 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. Four-weeks old mice (wild-type C57BL/6) were used for all experiments. Body weight gain and serum lipid levels were measured in the obesity-induced mice. Results DSH did not show toxicity even at the concentration of 1 mg/ml and DSH significantly inhibited the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner. Also, DSH significantly reduced the expressions of $PPAR{\gamma}$ and $C/EBP{\alpha}$ in a dose-dependent manner. Furthermore, DSH significantly reduced body weight gain, serum glucose, total cholesterol and LDL-cholesterol contents in obesity-induced mice. Conclusions These results demonstrated that DSH inhibited 3T3-L1 preadipocyte differentiations and high-fat diet-induced obesity in mice.

• Journal of Biomedical Engineering Research
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• v.39 no.3
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• pp.109-115
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• 2018

Obesity is a worldwide disease caused by the excessive proliferation of adipocytes. Multiple factors, including melatonin and physical loading, are involved in the control of obesity. Melatonin has been shown to induce apoptosis on preadipocytes while physical loading such as fluid shear stress (FSS) affects the proliferation and differentiation of adipocytes. Here, we studied the combined effects of melatonin and FSS on 3T3-L1 preadipocytes. For physical loading, preadipocytes were stimulated with a maximum dynamic fluid shear stress of 1 Pa at 1 Hz for 2 hours with/without melatonin. The experiment conditions were divided into four groups: (1) control, (2) 1 mM melatonin treatment, (3) FSS, and (4) combined 1 mM melatonin and FSS. All groups had a fixed duration time of 2 hours. ERK, p-ERK, COX-2, $C/EBP{\beta}$, $PPAR{\gamma}$, osteopontin, Bax, caspase-3 and caspase-8 proteins were assessed by Western blot analysis. GAPDH was used as a control. Results showed that combined melatonin and FSS treatment activated the ERK/MAPK pathway but not COX-2. Furthermore, combined melatonin and FSS treatment significantly decreased $C/EBP{\beta}$ and $PPAR{\gamma}$ compared to other groups. However, caspase-3 and caspase-8 did not result in significant changes. In summary, combined melatonin and FSS appears to have the potential to inhibit adipogenesis and treat obesity.

• BMB Reports
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• v.42 no.6
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• pp.338-343
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• 2009

The Wnt/$\beta$-catenin signaling pathway alters adipocyte differentiation by inhibiting adipogenic gene expression. $\beta$-catenin plays a central role in the Wnt/$\beta$-catenin signaling pathway. In this study, we revealed that tumour necrosis factor-$\alpha$ (TNF-$\alpha$), a potential negative regulator of adipocyte differentiation, inhibits porcine adipogenesis through activation of the Wnt/$\beta$-catenin signaling pathway. Under the optimal concentration of TNF-$\alpha$, the intracellular $\beta$-catenin protein was stabilized. Thus, the intracellular lipid accumulation of porcine preadipocyte was suppressed and the expression of important adipocyte marker genes, including peroxisome proliferator-activated receptor-$\gamma$ (PPAR$\gamma$) and CCAAT/enhancer binding protein-$\alpha$ (C/EBP$\alpha$), were inhibited. However, a loss of $\beta$-catenin in porcine preadipocytes enhanced the adipogenic differentiation and attenuated TNF-$\alpha$ induced anti-adipogenesis. Taken together, this study indicated that TNF-$\alpha$ inhibits adipogenesis through stabilization of $\beta$-catenin protein in porcine preadipocytes.

• Biomolecules & Therapeutics
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• v.18 no.1
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• pp.56-64
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• 2010

Pentoxifylline (PTX) has been used for the local reduction of fat tissue in the clinical setting. However, its safety and efficacy have not been proven. The aim of this study was to evaluate the effects of PTX on cell lines established from fat tissue. Newly cultured human preadipocytes and adipocytes from subcutaneous abdominal fat in addition to purchased human lung fibroblasts and keratinocytes were treated with PTX at different concentrations. Cell viability was determined using the Cell counting kit (CCK)-8 assay and lipolysis was evaluated using an Elisa kit. DNA fragmentation, Western blot analysis, Hoechst and Propidium Iodide (PI) staining and fluorescence activated cell scanning analysis were performed to confirm apoptosis. The viability of adipocytes, preadipocytes, keratinocytes and fibroblasts was markedly decreased at concentrations of PTX above 20 mM. Apoptosis was induced at concentrations of PTX over 40 mM in all cell lines. Lipolysis was increased by 60% at concentrations of PTX of 20 mM compared to the control. In conclusion, the results of this study showed that 20 mM of PTX induced lipolysis. At concentrations over 20 mM, PTX reduced the viability of all cells studied including: adipocytes, preadipocytes, fibroblasts and keratinocytes, in a non-specific manner.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.4-11
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• 2020

Objective: Puerarin has the potential of regulating the differentiation of preadipocytes, but its mechanism of action has not yet been elucidated. Adipocytes found in adipose tissue, the main endocrine organ, are the main sites of lipid deposition, and are widely used as a cell model in the study of in vitro fat deposition. This study aimed to investigate the effects of puerarin on adipogenesis in vitro. Methods: Puerarin was added to the culture medium during the process of adipogenesis. The proliferation and differentiation of bovine preadipocytes was measured through cell viability and staining with oil red O. The content of triacylglycerol was measured using a triglyceride assay kit. The mRNA and protein expression levels of adipogenic genes, peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer-binding protein-α, were measured using quantitative real-time polymerase chain reaction and western blotting, respectively. Results: The addition of puerarin significantly increased adipogenesis of bovine preadipocytes and enhanced the mRNA and protein level expression of PPARγ (p<0.01). The expression of P-Akt increased after adipogenic hormonal induction, whereas puerarin significantly increased PPARγ expression by promoting the Akt signaling component, P-Akt. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473, which may activate the downstream signaling of the Akt pathway. Conclusion: Puerarin was able to promote the differentiation of preadipocytes and improve fat deposition in cattle. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473.

• The Journal of Internal Korean Medicine
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• v.29 no.1
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• pp.80-89
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• 2008

Objectives : This study was carried out to investigate the effects of Ephedrae Herba pharmacopuncture (EHP) on the adipogenesis in 3T3-L1 cells, lipolysis in rat epididymal adipocytes and histological changes in porcine adipose tissue. Methods : Inhibition of preadipocyte differentiation and/or stimulation of lipolysis play important roles in reducing obesity. 3T3-L1 preadipocytes were differentiated with adipogenic reagents by incubating for 3 days in the absence or presence of EHP ranging from 0.01 to 1.0 $mg/m{\ell}$. The effect of EHP on adipogenesis was examined by measuring glycerol-3-phosphate dehydrogenase (GPDH) activity and by oil red O staining. Mature adipocytes from rat epididymal fat pad were incubated with EHP ranging from 0.01 to 1.0 $mg/m{\ell}$ for 3 days. The effect of EHP on lipolysis was examined by measuring free glycerol released. Fat tissue from porcine skin was injected with EHP ranging from 0.1 to 10.0 $mg/m{\ell}$ to examine the effect of EHP on histological changes under light microscopy. Results : The following results were obtained from present study on adipogenesis of preadipocytes, lipolysis of adipocytes and histological changes in fat tissue. Proliferation of preadipocytes was significantly inhibited by EHP at the concentration of 1.0 $mg/m{\ell}$. Lipolysis of adipocytes was increased by EHP at the concentration of 0.1, 1.0 $mg/m{\ell}$. Porcine fat tissues were widely injured by EHP at the concentration of 10.0 $mg/m{\ell}$. Conclusions : From the above results, EHP efficiently induces inhibition of preadipocytes proliferation, lipolysis of adipocytes and histologic injury in fat tissues. Therefore, EHP may be useful to treat localized obesity.

• Journal of Biomedical Engineering Research
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• v.42 no.1
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• pp.18-24
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• 2021

Vibration is a mechanical cue that can be applied to adipose tissues for the purpose of treating obesity. However, the exact correlation between vibration and other anti-adipogenic pathways, such as development of cytoskeleton and apoptosis, remains unknown. The objective of this study was to investigate the unknown anti-adipogenic effects of vibration with varied frequencies on 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. Vibration was generated using Arduino Uno microcontroller and vibration motor module with 1 V DC, and applied to preadipocytes for 3 days. Frequency conditions were set to 20, 55, and 90 Hz. Then, the expressions of p38 pathway, ROCK-1, α-actinin, Bax, Bcl-2, caspase-9, 8, and 3 were analyzed with western blot. As a result, p38 pathway was inhibited in 55 and 90 Hz while ROCK-1 and α-actinin were expressed in 20 Hz. Caspase-3, a terminal apoptotic factor, was activated in 20 Hz via extrinsic pathway rather than intrinsic pathway. Results suggest that various frequencies of vibration can inhibit adipogenesis via different pathways which sheds light on future mechanotransduction applications of vibration for the treatment of obesity.

• Journal of Marine Bioscience and Biotechnology
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• v.15 no.2
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• pp.59-66
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• 2023

Dictyopteris divaricata, a type of marine brown algae, has been studied for its various biological properties, including anti-inflammatory, antidiabetic, and whitening effects. However, its potential antiobesity effects have not been extensively explored. This study aimed to examine the impact of D. divaricata ethanol extract (DDE) on adipocyte differentiation and adipogenesis using 3T3-L1 preadipocytes. Our results showed that when 3T3-L1 preadipocytes were treated with noncytotoxic concentrations of DDE there was a concentration-dependent decrease in fat accumulation rate and triglycerid production compared with the control. Furthermore, DDE significantly reduced the expression of transcription factors (PPARγ, C/EBPα, and SREBP-1) and fatty acid transport protein (FABP4), which are crucial for 3T3-L1 preadipocyte differentiation. These findings suggest that DDE may exhibit antiobesity effects by suppressing the expression of lipogenic transcription factors and fatty acid transport proteins. Therefore, DDE holds potential as a therapeutic agent for obesity.