• Journal of Life Science
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• v.32 no.7
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• pp.523-531
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• 2022

The purpose of this study was to evaluate the effect of antioxidant and anti-adipogenic activities in ethanol extracts from herb mixture (Ephedra sinica, Atractylodes lance, Gypsum fibrosum, and Theobroma cacao). DPPH, ABTS⁺ radical and xanthine oxidase scavenging activities were measured for antioxidant activity. Extracts of the herb mixture had 75.0, 100.8, and 79.5% scavenging activities at 1,000 ㎍/ml concentration, respectively. We investigated the inhibition of adipogenesis and adipocyte differentiation with an extract of an herb mixture in 3T3-L1 preadipocytes. An extract from the herb mixture at concentrations between 0 and 50 ㎍/ml did not affect 3T3-L1 cell viability. Treatment with herb mixture extracts of 25, 50, and 75 ㎍/ml in 3T3-L1 preadipocytes inhibited lipid accumulation in a dose-dependent manner. As a result of a Western blot experiment, it was shown that the herb mixture inhibited the differentiation transcription factors, PPARγ and C/EBPa, by 44.2 and 77.6%, respectively, at a concentration of 75 ㎍/ml in MDI-induced differentiated 3T3-L1 cells. As a result of RT-PCR, the gene expression of C/EBPa, SREBP-1c, and PPARγ was significantly inhibited by 43.4%, 59.6%, and 55.3%, respectively, at the concentration of 75 ㎍/ml of the herb mixture compared with the MDI-treated group. In addition, the expression of fatty acid synthase (FAS), a fatty acid synthesis regulator, was suppressed. These results can be applied to develop a functional food for anti-obesity with a herb mixture.

• Journal of Life Science
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• v.21 no.5
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• pp.720-728
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• 2011

Garlic (Allium sativum) has been used as a source food as well as a traditional folk medicine ingredient since ancient times. Aged black garlic is a type of fermented garlic and is expected to have stronger anticancer and antioxidant activities than raw garlic. However, the mechanisms of their inhibitory effects on adipocyte differentiation and adipogenesis are poorly understood. In the present study, the effects and mechanisms of water extracts of raw garlic (WERG) and aged black garlic (WEABG) on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes were investigated. Treatment with WEABG significantly suppressed terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner as confirmed by a decrease in lipid droplet number and lipid content through Oil Red O staining, however WERG had no such effect. In addition, WEABG reduced accumulation of cellular triglyceride, which is associated with a significant inhibition of key pro-adipogenic transcription factors including peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding proteins ${\alpha}$ (C/EBP${\alpha}$) and C/EBP${\beta}$. Taken together, these results provide important new insight that aged black garlic might inhibit adipogenesis by suppressing the pro-adipogenic transcription factors in 3T3-L1 preadipocytes, and further studies will be needed to identify the active compounds that confer the anti-obesity activity of aged black garlic.

• Journal of the Korea Society of Computer and Information
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• v.29 no.1
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• pp.187-194
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• 2024

In this study, we examined the effects of Rubus crataegifolius leaf on the inhibition of differentiation and adipogenesis of 3T3-L1 preadipocytes to confirm their potential for use as an anti-obesity functional material. Rubus crataegifolius leaves water extracted using hot water were then concentrated for use, with an extract yield of 4.76%. The result of measuring the rate of 3T3-L1 cell survival of Rubus crataegifolius leaf extract (RCLE) showed growth inhibition of 13% at a concentration of 1,000 ㎍/mL. Thus, in this study, experiments were performed using RCLE treatment concentrations up to 500 ㎍/mL. Production of triglycedie in 3T3-L1 cells showed a dose-dependent decrease, and the rate of reduction was 28.7, 40.8, and 51.6% at concentrations of 100, 300, and 500 ㎍/mL, respectively, compared to the control group. In addition, the results confirmed that suppression of lipogenesis was achieved by suppressing the expression of peroxisome proliferator-activated receptor γ (PPAR γ), CCAAT/enhancer-binding protein α (C/EBP α), sterol regulatory element-binding protein-1c (SREBP-1c), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), and increasing the expression of p-activated protein kinase (p-AMPK). Based on these results, it is believed that Rubus crataegifolius leaf extract can be used in the effort to manage obesity by regulating factors related to adipocyte differentiation and adipogenesis.

• YAKHAK HOEJI
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• v.38 no.3
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• pp.238-249
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• 1994

The purpose of this research was to investigate effects of baicalin on the differentiation of preadipocytes, 3T3-L1, and to characterize the action of baicalin that affect the responses of 3T3-L1 cells during differentiation. In various culture conditions, effects of baicalin and adrenoreceptor agonists such as phenylephrine(PE) and isoproterenol(IPR) on cell differentiation were examined. Also, effects of the drugs on differentiation, triglyceride(TG) contents, expression of insulin receptor, cAMP contents, the cytosolic $Ca^{2+}$ levels, and amount of calmodulin(CaM) were examined. The results suggest that baicalin has adrenergic receptor blocking activity during the process of differentiation of 3T3-L1 cells and that in the early stage of the adipose conversion, the effect of baicalin on the adipocyte differentiation is mediated by the regulation of insulin receptor expression, but by alterations of the cAMP and the calcium metabolism in the late stage. These results also suggest that the action of baicalin may be significant in the lipid metabolism, lipogenic and lipolytic pathways, of adipose cells.

• Journal of Life Science
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• v.25 no.1
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• pp.62-67
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• 2015

Myelophycus simplex Papenfuss is distributed over the northern Pacific and southern coast of Korea, and is a member of the brown algae family. The objective of this study was to investigate the effect of M. simplex methanol extract on adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes. Treatment with M. simplex methanol extract significantly suppressed terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in lipid droplet content observed by Oil Red O staining. Also, the M. simplex methanol extract significantly suppressed the triglyceride content of 3T3-L1 preadipocytes in a dose-dependent manner. Treatment with 300 and $500{\mu}g/ml$ of M. simplex methanol extract caused a 42% and 76% reduction in lipid droplet content, respectively. In order to understand the anti-adipogenic effects of M. simplex methanol extract, the changes in the expression of several adipogenic transcription factors, including peroxisome proliferator-activated receptor (PPAR) ${\gamma}$-cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding protein (C/EBP) ${\alpha}$ and ${\beta}$, were investigated using immunoblotting. M. simplex suppressed the expression of $PPAR{\gamma}$, $C/EBP{\alpha}$, and $C/EBP{\beta}$ proteins compared with control. Therefore, the results of this study suggest that M. simplex methanol extract inhibits adipocyte differentiation and thus may have applications as a potential source for an anti-obesity functional food agent.

• YAKHAK HOEJI
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• v.55 no.6
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• pp.478-484
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• 2011

Non-alcoholic fatty liver disease is known to be frequently associated with obesity and type 2 diabetes. We examined the effects of EtOH extracts from Triticum aestivum on lipid accumulation during the differentiation of 3T3-L1 preadipocytes to screening the candidate materials in preventing non-alcoholic fatty liver disease. The lipid level in adipocytes was determined by Oil Red O staining. The treatment of 50% ethanol, but not water and 100% ethanol extracts, from Triticum aestivum at concentration of 0.5 $mg/ml$ inhibited lipid accumulation in 3T3-L1 cells, revealing no cell toxicity. Thus, the fractions of $CH_2Cl_2$, EtOAc and BuOH were separated from 50% EtOH extract to characterize anti-adipogenic effect. The $CH_2Cl_2$ fraction at concentration of $50{\mu}g/ml$ effectively inhibited the lipid accumulation in the adipocytes compared to those of EtOAc and BuOH at concentration of $50{\mu}g/ml$. The intracellular triglyceride accumulation also was significantly reduced by treatment of $CH_2Cl_2$ fraction in concentration-dependent manner. Western blot analysis showed that the $CH_2Cl_2$ fraction attenuated the intracelluar level of fatty acid synthase(FAS) accompanied by attenuated expression of Peroxidase proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$) adipogenic transcription factor. These results suggest that $CH_2Cl_2$ fraction from 50% EtOH extract of Triticum aestivum may has the potent anti-adipogenic effects by inhibiting the transactivation of $PPAR{\gamma}$.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.5
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• pp.1315-1320
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• 2003

The purpose of this research was to investigate effects of Bambusae Caulis in Liquamen (BCL) on the synthesis of basement membrane proteins during proliferation and differentiation of 3T3-L1 cells. BCL has been used to relieve the cough and asthma, and remove phlegm in traditional oriental medicines. In recent years. it was studied for its antiinflammatory, antiallergenic. immune-modulating and anticarcinogenic capabilities. We have previously observed that glycyrrhizin stimulates the adipose conversion of 3T3-L1 cells. To investigate effects of BCL on the basement membrane proteins during proliferation and differentiation of 3T3-L1 cells, we have analyzed synthetic amounts of basement membrane components such as type IV collagen and BM40. BCL stimulated the synthesis and secretion of type IV collagen from both 3T3-L1 preadipocytes and adipocytes. The synthesis and secretion of BM40 was not affected by BCL. The continuous addition of BCL markedly stimulated cell growth and increased cell density. These results suggest an important role for type IV collagen in adipocyte differentiation.

• Journal of Pharmacopuncture
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• v.12 no.2
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• pp.13-20
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• 2009

In this study, the effects of Ganoderma lucidum (GL) on fat metabolism were performed in 3T3-L1 adipocytes. The effects of GL on 3T3-L1 preadipocytes differentiation were also examined. Our results showed that GL decreased the TG content by ORO staining. To elucidate the mechanism of the effects of GL on lowering TG content in 3T3-L1 adipocytes, we examined whether GL modulate the expressions of transcription factors and adipokines related to control of energy expenditure process because adipokines regulate adipocyte mass and increased expenditure may consume much TG in adipocytes. As a result, the expression of C/$EBP{\beta}$, C/$EBP{\delta}$, C/$EBP{\alpha}$, and $PPAR{\gamma}$, genes, which induce the adipose differentiation and adipose-specific FAS, aP2, and adipsin genes, which compose fat formation were decreased. In addition, GL increased the expression of leptin, UCP2, adiponectin in 3T3-L1 adipocytes, resulting in energy homeostasis. In conclusion, GL could regulate transcript factor related to induction of adipose differentiation and control TG content by up-regulation of adipokines related to fat burn.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.1
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• pp.119-125
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• 2006

The present study including two experiments was designed to determine the effect of media containing different rare earth elements (REE) on proliferation and fatty acids accumulation in 3T3-L1 cell cultures. In Experiment 1, 3T3-L1 preadipocytes in 96-well plates ($1.5{\times}10^4cells/ml$) were cultured with Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) for 24 h. Then the media were changed to the following 10 different media for 48 h: DMEM containing 10% FBS for the control; the above media containing $5{\mu}M$, $10{\mu}M$ or $15{\mu}M$ of $LaCl_3$, $CeCl_3$ or the mixture of these REE chlorides. The proliferation rate of the cells was measured and compared by a non-isotope method-XTT method. In Experiment 2 the cells in 24-well plates ($1.5{\times}10^4cells/ml$) were cultured in DMEM containing 10% FBS for 7 days until confluent and then were changed to above DMEM containing dexamethasone, methyl-isobutylxanthine and insulin (DMI) for two days. Afterwards the media were changed to the 10 different media with REE supplements as in Experiment 1 and cultured for 6 days. The cells were then harvested for fatty acids analysis by gas chromatography. It was found that supplementation of La (5, 10 and $15{\mu}M$), Ce ($5{\mu}M$ and $15{\mu}M$) and the mixture REE (5, 10 and $15{\mu}M$) stimulated (p<0.05) the proliferation of 3T3-L1 preadipocytes (Experiment 1). In the differentiating 3T3-L1 cells supplementation of La ($5{\mu}M$ and $10{\mu}M$), Ce ($5{\mu}M$) and the mixture REE ($5{\mu}M$ and $15{\mu}M$) decreased (p<0.05) the concentration of monounsaturated fatty acids (MUFA) per $10^5cells$, while the supplementation of La ($5{\mu}M$), Ce ($5{\mu}M$) and the mixture REE ($15{\mu}M$) increased (p<0.05) the ratio of saturated fatty acids (SFA) to MUFA. These results indicate that the supplementation of REE to the media may affect proliferation, differentiation and lipogenesis rates of 3T3-L1 cells. However, the effect may depend upon the level or type of REE applied.

• Journal of Life Science
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• v.29 no.1
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• pp.60-68
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• 2019

Limonium tetragonum, an edible halophyte that grows on salt marshes in Korea, is thought to possess various health benefits (e.g., antioxidant, antitumor, and hepatoprotective). In the present study, different solvent partitioned subfractions, water ($H_2O$), buthanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and hexane (n-hexane), from crude extract of L. tetragonum were tested for their ability to prevent adipogenesis in differentiating 3T3-L1 preadipocytes. The treatment of differentiating 3T3-L1 preadipocytes with L. tetragonum subfractions (LTFs) resulted in suppressed adipogenesis and reduced expression of adipogenesis-related transcription factors such as peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), CCAATT/enhancer-binding protein alpha ($C/EBP{\alpha}$), and sterol regulatory element-binding protein 1c (SREBP-1c) at both mRNA and protein levels. In addition, the LTF treatment notably decreased the levels of phosphorylated p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) of the mitogen-activated protein kinase (MAPK) pathway in association with $PPAR{\gamma}$-linked adipogenesis. Among all the tested LTFs, $H_2O$ and n-hexane were the most effective in lowering lipid accumulation and regulating the adipocyte differentiation via $PPAR{\gamma}$ pathway. Taken together, the results indicated that the $H_2O$ and n-hexane LTFs contain bioactive compounds that may exhibit significant antiadipogenesis activity by downregulation of the $PPAR{\gamma}$ pathway and inactivation of the MAPK signal pathway in 3T3-L1 preadipocytes.