• Journal of Korean Medicine for Obesity Research
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• v.23 no.2
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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.

• Journal of Animal Science and Technology
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• v.62 no.6
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• pp.854-863
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• 2020

This study was aimed to investigate the inhibitory effects of Porphyra dentata (P. dentata) extract on the adipogenesis of 3T3-L1 cells and evaluate its anti-obesity effect. The proliferation of 3T3-L1 cells and differentiation of adipocytes under treatment of P. dentata extract was examined by measuring the cell viability using alamarBlue assay and lipid droplets by Oil Red O staining. Results showed that P. dentata extract has no cytotoxicity effect and lipid droplets formation decreased in a concentration-dependent manner in 3T3-L1 cells. It has been confirmed that transcription factors affecting lipid accumulation and anti-adipogenic effects during cell differentiation are linked to P. dentata extract. We observed that P. dentata shows lowering the mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα) that adipogenesis-associated key transcription factors and inhibiting adipogenesis in the early stages of differentiation. Treating the cells with P. dentata did not only suppressed PPARγ2 and C/EBPα but also significantly decreased the mRNA expression of adiponectin, Leptin, fatty acid synthase, adipocyte protein 2, and Acetyl-coA carboxylase 1. Overall, the P. dentata extract demonstrated inhibitory property in adipogenesis, which has a potential effect in anti-obesity in 3T3-L1 cells.

• Nutrition Research and Practice
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• v.17 no.6
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• pp.1043-1055
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• 2023

BACKGROUND/OBJECTIVES: The fruit of Cydonia oblonga Miller (COM) is used traditionally in Mediterranean region medicine to prevent or treat obesity, but its mechanism of action is still unclear. Beyond a demonstrated anti-obesity effect, the fruit was tested for the mechanism of adipogenesis in 3T3-L1 preadipocytes. MATERIALS/METHODS: 3T3-L1 preadipocytes were cultured for 8 days with COM fruit extract (COME) at different concentrations (0-600 ㎍/mL) with adipocyte differentiation medium. The cell viability was measured using an MTT assay; triglyceride (TG) was stained with Oil Red O. The expression levels of the adipogenesis-related genes and protein expression were analyzed by reverse transcription polymerase chain reaction and Western blotting, respectively. RESULTS: COME inhibited intracellular TG accumulation during adipogenesis. A COME treatment in 3T3-L1 cells induced upregulation of the adenosine monophosphate-activated protein kinase (AMPK)α phosphorylation and downregulation of the adipogenic transcription factors, such as sterol regulatory element-binding protein 1c, peroxisome proliferator-activated receptor γ, and CCAAT/enhancer binding protein α. The COME treatment reduced the mRNA expression of fatty acyl synthetase, adenosine triphosphate-citrate lyase, adipocyte protein 2, and lipoprotein lipase. It increased the mRNA expression of hormone-sensitive lipase and carnitine palmitoyltransferase I in 3T3-L1 cells. CONCLUSIONS: COME inhibits adipogenesis via the AMPK signaling pathways. COME may be used to prevent and treat obesity.

• Journal of Korean Medicine for Obesity Research
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• v.19 no.2
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• pp.89-96
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• 2019

Objectives: We investigated whether membrane free stem cell extract from adipose tissue (MFSCE) has anti-diabetic effect. Methods: To determine glucose uptake effect of MFSCE, we carried out glucose uptake assay in 3T3-L1 adipocytes. The regulatory mechanisms of MFSCE on glucose uptake were examined by Western blot analysis. Results: When MFSCE was treated to adipocytes at the concentration of 0.5, 1, 2.5, and 5 ㎍/mL, 2-deoxyglucose-6-phosphate uptake was elevated approximately 1.8-fold compared to cells not treated with MFSCE. It indicated that MFSCE enhances glucose uptake in 3T3-L1 adipocytes. In addition, MFSCE reduced phosphorylation of insulin receptor substrate-1 at serine 307 and induced Akt and glucose transporter 4 protein expressions that were related to insulin signaling. Furthermore, MFSCE regulated adenosine monophosphate-activated protein kinase (AMPK) pathway by increases of increase phosphorylation of AMPK and acetyl-CoA carboxylase that were related to AMPK pathway. Conclusions: These results indicated that MFSCE promotes glucose uptake via modulation of insulin signaling and AMPK pathway. Therefore, MFSCE could be a promising agent for treatment of diabetes mellitus.

• Journal of Korean Medicine for Obesity Research
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• v.22 no.1
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• pp.38-46
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• 2022

Objectives: Juknyeok (JN) is natural liquor extracted from bamboo stems (Phyllostachys bambusoides) and has been used as a traditional Korean medicine for improving vascular function, blood glucose, and treating stroke. Until now, the JN's lipid-lowering effect and underlying mechanism in adipocytes are poorly understood. The aim of this study was to scrutinize the effect of a standardized commercial JN on lipid accumulation during the differentiation of 3T3-L1 preadipocytes. Methods: Lipid and triglyceride (TG) accumulation in differentiating 3T3-L1 preadipocytes were measured by Oil Red O staining and AdipoRed assay, respectively. Cell count analysis was used to ascertain 3T3-L1 cytotoxicity. Immunoblotting and Reverse transcription polymerase chain reaction analysis were used to assess protein and messenger RNA (mRNA) expression levels in 3T3-L1 cells, respectively. Results: Treatment with JN at 25 𝜇l/ml after pH calibration with 6.35 significantly reduced lipid and TG accumulation in differentiating 3T3-L1 preadipocytes without significant cytotoxicity. On mechanistic levels, JN markedly suppressed protein expression levels of CCAAT/enhancer-binding protein (C/EBP)-𝛽 and fatty acid synthase (FAS) during the differentiation of 3T3-L1 preadipocytes. However, JN did not affect the protein expression levels of C/EBP-𝛼, peroxisome proliferator-activated receptor-𝛽/𝛾, and phosphorylation levels of signal transducer and activator of transcription-3/5 in differentiating 3T3-L1 preadipocytes. JN also reduced leptin mRNA expression levels in differentiating 3T3-L1 preadipocytes. Conclusions: JN at 25 𝜇l/ml lowers lipid accumulation and TG content in differentiating 3T3-L1 cells, mediated through the reduced expression levels of C/EBP-𝛽 and FAS.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.1
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• pp.32-38
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• 2009

In this study, we investigated the anti-obesity activity of the herbal extract mixture (HEM). The inhibitory effect of HEM on triglyceride accumulation of 3T3-L1 preadipocyte was examined by Oil-Red O staining. HEM inhibited the triglyceride accumulation of 3T3-L1 preadipocyte cell and reduced glycerol-3-phosphate dehydrogenase (GPDH) activity. We further investigated the effect of HEM in prevention of obesity in male ICR mouse for 5 weeks. Experimental groups were divided into high fat diet group (HFD), HFD supplemented with 100 mg/kg HEM group (HEM1) and HFD supplemented with 200 mg/kg HEM group (HEM2). Body weight and food efficiency ration of HEM1 and HEM2 was decreased by 52% and 50% and by 45% and 50%, respectively. The amount of adipocyte in body weight was decreased. Blood triglyceride and total cholesterol of HEM1 was significantly decreased. These results indicate that HEM may be useful in preventing obesity.

• Journal of Life Science
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• v.18 no.11
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• pp.1600-1605
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• 2008

This study was performed to verify the gene expression of 3T3-L1 using the siRNA of the aromatase gene, which is the estrogen synthesis enzymes. First of all three pairs of siRNA were designed from the CYP19A1 (aromatase) and analyzed the formation of fat cell mechanism by transferring gene to 3T3-L1 and differentiating it. As a result, the expression of leptin gene, which is the main gene causing the obesity, was controlled and the cause of the obesity is related with the insulin specifically. The overexpression of adiponectin and adipsin was observed. This result showed that the formation of the fat was controlled a little without any side effect by obstructing a specific material out of all the signal systems in the fat formation. This study will be an important clue to make it clear that the lack or overexpression of estrogen might be the cause of fat formation mechanism.

• Journal of Life Science
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• v.30 no.10
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• pp.835-843
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• 2020

Obesity, the world's leading metabolic disease, is a serious health problem in both industrialized and developing countries. Natural substances are of great interest in preventative medicine, especially in the field of metabolic syndromes-from insulin resistance to obesity and diabetes. In the present study, we investigated the effect of A. pullulans SM-2001 Extract (Polycan^®) on the adipocyte differentiation of 3T3-L1 preadipocytes and the anti-obesity effect of 3T3-L1 adipocytes. Although β-glucan has been found to have health benefits in the regulation of the immune system and blood cholesterol levels, its role in obesity has not been fully investigated. Polycan^® suppressed lipid accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity without affecting cell viability in 3T3-L1 preadipocytes and adipocytes. Polycan^® also inhibited cellular lipid accumulation through down-regulation of transcription factors, such as PPARγ and C/EBPα, and induced dose-dependent phosphorylation of AMP-activated protein kinase (AMPK)-a cellular energy sensor-while the total AMPK protein content remained unchanged. Taken together, this shows that the activation of AMPK by Polycan^® in adipocytes plays a critical role in Polycan^®-induced inhibition of adipocyte differentiation. Our results show that Polycan^® has an anti-obesity action in vitro, suggesting a potential novel preventative agent for obesity and other metabolic diseases.

• Korean Journal of Oriental Medicine
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• v.9 no.2
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• pp.45-54
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• 2003

Obesity is caused by unbalance of energy intake and expenditure, which results in extra accumulation of adipose tissue. Obesity is directly related to metabolic diseases such as diabetes, hyperlipidemia, fatty liver and so on. To investigate the anti-obesity effects of Scutellariae Radix, 70% EtOH extract and water extract of it were tested by in vitro and in vivo studies of fat accumulation. 3T3-L1 preadipocyte cell line was used in a in vitro study of fat accumulation. After 3T3-L1 cells were induced to differentiate into adipocytes, S. radix extract were added and fat accumulation was measured by oil red O staining. In vivo study showed that weight and epididymal/ retro-peritoneal adipose tissues were significantly reduced in mice fed Scutellariae Radix extract compared with control group. Especially, mice fed Scutellariae Radix extract showed reduced serum triglyceride and glucose levels. When adipose tissues were analyzed by microscope, mean adipocyte size was significantly reduced in Scutellariae Radix extract-fed mice. Therefore, this study showed inhibitory effects of Scutellariae Radix on in vitro and in vivo fat accumulation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.6
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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.