• Korean Journal of Clinical Laboratory Science
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• v.51 no.4
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• pp.514-520
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• 2019

This study investigated the improved lipid metabolism effect of 3T3-L1 cells induced by adipocytes using the dichloromethane (DCM) fraction in the organic solvent extract of Wassong (Orostachys japonicus). To confirm the cell cytotoxicity, each of 6 fractions of organic solvent extracts (EtOH, Hexane, DCM, EtOAc, BuOH, and H₂O) was examined using MTS assay. As a result, it was confirmed that the DCM extract was stable over the whole range of concentrations, and a DCM fraction was used to confirm the improved lipid metabolism effect. Lipid excretion was measured to confirm the change of lipid metabolism. 3T3-L1 cells induced by adipocytes were treated with DCM extract and stained with oil-red O to evaluate lipid accumulation. As a result, it was confirmed that the lipid efflux was significantly improved. In order to confirm the mechanism of lipid efflux, the mRNA expressions of ABCA1 and ABCG1, which are lipid transport proteins, were confirmed by real-time PCR. Therefore, the present study confirmed that the DCM extract from Orostachys japonicus has the effect of improving the lipid metabolism on 3T3-L1 adipocytes. In addition, the results of this study will be used as the basis for the development of functional foods using Orostachys japonicus and also for conducting research on the detailed mechanisms.

• Nutrition Research and Practice
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• v.7 no.6
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• pp.453-459
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• 2013

The abnormal content of blood lipids often results in metabolic diseases, such as hyperlipidemia and obesity. Many agents, including natural sources from traditional food, have been developed to regulate the blood lipid contents. In this study, we examined the anti-hyperlipidemic activity of Rhynchosia nulubilis seeds pickled with brown rice vinegar (RNSpBRV), a Korean traditional pickled soybean food. Since RNSpBRV is made of R. nulubilis seeds (RNS) soaked in brown rice vinegar (BRV), we compared the anti-adipogenic activity between RNS, BRV and solid fraction of RNSpBRV (SF-RNSpBRV), liquid fraction of RNSpBRV (LF-RNSpBRV). For this, the inhibitory effect of lipid accumulation in 3T3-L1 adipocyte was checked by adding methanol extracts of mixed RNS and BRV, LF-RNSpBRV, and SF-RNSpBRV. The addition of each methanol extract up to 1 mg/ml showed no cytotoxicity on 3T3-L1 adipocyte, and approximately 20% of the lipid droplet formation was suppressed with the methanol extract of BRL or SF-RNSpBRV. The highest suppression (42.1%) was achieved with LF-RNSpBRV. In addition, mice fed a high fat diet (HFD) supplemented with 5% RNSpBRV powder led to increased high density lipoprotein (HDL) cholesterol and lower blood glucose, triglyceride, and total cholesterol compared to mice fed with a HFD diet only. Interestingly, the size of the epididymis cells gradually decreased in HFD + 1% RNSpBRV and HFD + 5% RNSpBRV-fed mice if compared those of HFD-fed mice. Taken together, these results provide evidence that RNSpBRV has a regulatory role in lipid metabolism that is related to hyperlipidemia.

• 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.

• Food Science and Preservation
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• v.22 no.5
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• pp.744-750
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• 2015

The present study was designed to investigate the effects of hot water and ethanol extracts of Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with both hot water and ethanol extracts for up to 8 days following standard induction of differentiation. Regarding anti-adipogenic activity, compared with the control, the hot water and ethanol extracts significantly inhibited lipid accumulation (37.4 and 66.6%, respectively) and ROS production (46.4 and 46.8%, respectively) during adipogenesis in 3T3-L1 cells. Treatment with hot water and ethanol extracts significantly inhibited mRNA expression of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), thereby reducing the mRNA expression of adipocyte-specific fatty acid binding protein (aP2). Moreover, both the extracts significantly inhibited mRNA expression of NADPH oxidase (NOX4). Overall, our research suggests that N. nucifera Gaertner flower extracts might be a valuable source of bioactive compounds that exhibit anti-adipogenic activity and could have applications in the field of medicine and food industry.

• Journal of Life Science
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• v.26 no.3
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• pp.325-330
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• 2016

In this study, we evaluated the antidiabetic effect of submerged culture of Ceriporia lacerata mycelium (CL01) on glucose uptake and the expression of mRNA and protein of major signal markers of insulin signaling pathway in 3T3-L1 adipocytes. After 3T3-L1 adipocytes were pre-treated by CL01 (0, 2, 10 mg/ml) for 8 hours, followed with treatment of insulin, the glucose uptake levels significantly increased by more 55.1%, 94.4% than negative control respectively (p<0.01, 0.001) in a dose-dependent manner. However, in case of CL01 pre-treatment without insulin, the glucose uptake did not increase compared with insulin-treated 3T3-L1. Also we demonstrated that the protein expression levels of pIR β, pAkt, pPI3K and pAMPK and the mRNA expression levels of GLUT4 in adipocytes inducing insulin resistance increased in CL01-treated group compared with negative control. These results demonstrated that CL01 affected glucose metabolism and the protein and gene expression through insulin signaling pathway, and increased glucose uptake levels effectively. More than 90% of those who have suffered for type 2 diabetes are more likely to have from hyperinsulinemia, hypertension, obesity and etc. because of altered insulin signaling pathway. So, it is probably considered that intake of CL01 may treat type 2 diabetes by normalization of insulin signaling pathway, and it will provide useful evidences regarding a mechanism for cure of type 2 diabetes.

• Journal of Nutrition and Health
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• v.52 no.1
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• pp.17-25
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• 2019

Purpose: Obesity is a major health problem of global significance because it is clearly associated with an increased risk of health problems, such as nonalcoholic fatty liver disease (NAFLD), diabetes, cardiovascular diseases, and cancer. Lonicera caerulea (LC) originates from high mountains or wet areas and has been used as a traditional medicine in northern Russia, China, and Japan. LC contains a range of bioactive constituents, such as vitamins, minerals, and polyphenols. This study examined the anti-obesity effects of LC during differentiation in preadipocytes. Methods: The cell viability assay was performed after the differentiation of 3T3-L1 cells for 7 days. Oil Red O staining was used to visualize the changes in lipid droplets in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs). The mRNA expression of obesity-related genes was determined by quantitative real-time PCR. Results: According to the results of Oil Red O staining, the lipid levels and size of lipid droplets in the adipocytes were reduced and the LC extract (LCE, 0.25-1 mg/mL) markedly inhibited adipogenesis in a dose-dependent manner. The treatment of LCE also decreased the mRNA expression of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), CCAAT/enhancer binding protein-${\alpha}$ ($C/EBP{\alpha}$), and sterol regulatory element binding protein 1 (SREBP1) in 3T3-L1 cells. Western blot analysis showed that the $PPAR{\gamma}$, $C/EBP{\alpha}$, and SREBP1 protein levels in both 3T3-L1 and MADSC were reduced in a dose-dependent manner. Conclusion: These results suggest that LCE can inhibit adipogenic differentiation through the regulation of adipogenesis-related markers.

• The Korea Journal of Herbology
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• v.29 no.2
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• pp.15-21
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• 2014

Objectives : The purpose of this study was to investigate inhibitory effects of steamed Polygonatum odoratum extract (POE) on differentiation and adipogenesis in 3T3-L1 adipocytes. Methods : Polygonatum odoratum (P. odoratum) extract was extracted with ethyl acetate. Total phenolic and flavonoid contents in POE were measured for antioxidant activity. The spectrophotometric method was used to determine the DPPH and ABTS radical scavenging activity and ferric-reducing antioxidant potential (FRAP). MTT assay was examined for cell toxicity, oil red O staining was performed for intracelluar adipogenesis in differentiated 3T3-L1 adipocytes. Western blot analysis for measurement of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor${\gamma}$ ($PPAR{\gamma}$) and AMP-activated protein kinase (AMPK) expressions were performed. Results : The results revealed that POE has antioxidant activities. Contents of total polyphenolics and flavonoids were $50.83{\pm}1.52$ GAE mg/100g dry weight of POE and $17.05{\pm}2.47$ RE mg/100g dry weight of POE, respectively. DPPH radical scavenging activity, and FRAP in 10 mg/ml concentration were $92.1{\pm}0.6%$, $244.8{\pm}9.0{\mu}M$ Fe(II) and ABTS inhibition in 5 mg/ml concentration was $84.8{\pm}4.1%$. Treatment of POE in adipocytes inhibited the differentiation and adipogenesis of 3T3-L1 adipocytes compared to those of vehicle control. Additionally, protein expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, major transcription factor for the adipogenic genes, were significantly decreased compared to those of vehicle control (p<0.05). Futhermore, phosphorylation of AMPK was increased in 3T3-L1 adipocytes treated with POE compared to that of vehicle control (p<0.05). Conclusions : we demonstrate that steamed P. odoratum extract (POE) has potentiating antioxidant activities, inhibits differentiation and lipid accumulation and also induces energy expenditure in adipocytes, which may contribute to antiobesity property.

• Food Science of Animal Resources
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• v.39 no.3
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• pp.430-445
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• 2019

Natural edible waxes mixed with plant oils, containing high levels of unsaturated fatty acids (FAs), are known as oleogels. Oleogels are used for replacing saturated FAs in animal-derived food with unsaturated FAs. However, the health effects of edible waxes are not yet clearly defined. The purpose of this study was to investigate the effect of FAs and natural waxes on the adipogenesis in 3T3-L1 cells. The 3T3-L1 cells were differentiated and treated with FAs and waxes. These FAs [Palmitic acid (PA), Stearic acid (SA), Oleic acid (OA), Linoleic acid (LA), and Alpha-linolenic acid (ALA)] and waxes [beeswax (BW) and carnauba wax (CW)] were prepared at varying concentrations, and cell toxicity, triglyceride accumulation, lipid droplets size, and distribution inside of cells were determined. Adipogenic gene expression including $PPAR{\gamma}$, FASN, $C/EBP{\alpha}$, SREBP-1, and CPT-1 was determined. Results showed that increasing the concentration of FAs and waxes led to a decrease in the adipocyte cells viability and metabolic performance. SA showed the highest level of triglyceride accumulation (p<0.05), whereas ALA showed the lowest (p<0.05). Both BW and CW at 3.0 ppm showed significantly higher lipid accumulation than in the control and other groups (p<0.05). ALA had significantly downregulated adipogenic gene expression levels, excluding those of CPT-1, compared to the other treatment groups (p<0.05). Moreover, BW demonstrated similar adipogenic gene expression levels as ALA compared to CW. Consequently, ALA and BW may have health benefits by reducing adipogenesis and can be used in processed meat.

• The Korean Journal of Food And Nutrition
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• v.25 no.4
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• pp.800-806
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• 2012

Reactive oxygen species (ROS) are produced by oxidative stresses which cause various chronic diseases such as diabetes and obesity. Ginseng (Panax ginseng C.A. Mayer) has been reported to contain various biological activities such as anti-cancer, anti-diabetic, neuroprotective, radioprotective, anti-amnestic and anti-aging effects. In this study, we investigated the effects of Panax ginseng, treated with high temperatures and high pressures, on oxidative stress in C2C12 myoblasts and 3T3-L1 adipocytes. Oxidative stress was induced in the C2C12 cells through the introduction of $H_2O_2$ (1 mM), and cells were then treated with various ginseng preparations: dried white ginseng (DG), steamed ginseng (SG) and high temperature and high pressure treated ginseng (HG). In addition, 3T3-L1 preadipocytes were treated with various ginsengs for up to 8 days following standard induction of differentiation. Our results show that HG treatment significantly protected oxidative stress in both cell lines and enhanced gene expression of antioxidant enzymes. Therefore, in this study, we investigated the protective effects of ginseng on the oxidative stress of adipocytes and muscle cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.9
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• pp.1264-1269
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• 2015

To evaluate the functional effect of ornithine produced by isolated lactic acid bacteria, we examined the anti-lipid accumulation effect of ornithine produced by isolate lactic acid bacteria on 3T3-L1 cells. Lactic acid bacteria (Pediococcus strain) were isolated from nuruk, which is made from wheat, rice, and barley (whole grain, grits, or flour) by fermenting microorganisms (Aspergillus, Rhizopus, and yeasts). Pediococcus strain was identified by 16S rDNA sequencing analyses, and cells were collected by centrifugation and developed as an ornithine starter. makgeolli, an ornithine-containing Korean traditional alcoholic beverage, was made with isolated lactic acid bacteria and arginine. makgeolli was made with the help of ornithine starter using a makgeolli making kit. We evaluated the anti-proliferation effect of ornithine makgeolli on 3T3-L1 cells. To determine the anti-proliferation effect of ornithine makgeolli on preadipocytes, lipid droplets were quantified and stained with Oil Red O. makgeolli made with ornithine starter and arginine showed a 3-fold higher concentration of ornithine compared to makgeolli without starter and arginine. In the results of 3T3-L1 cell line experiment, lipid accumulation was significantly reduced by adding 0.05 mg/mL of ornithine makgeolli compare to the control (adipocyte without sample). In conclusion, ornithine makgeolli containing ornithine starter isolated from nuruk showed an anti-lipid accumulation effect with increased ornithine content without toxicity.