• Preventive Nutrition and Food Science
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• v.11 no.2
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• pp.110-114
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• 2006

In order to develop the improved kochujang witb an anti-adipogenic effect, garlic-added kochujang was prepared and followed by fermentation at $30^{\circ}C$ for 120 days. Anti-adipogenic effects of the garlic-added kochujang in cultured 3T3-L1 adipocytes were investigated by measuring leptin and glycerol levels as indicators of lipid accumulation associated with lipolysis, respectively. Additional levels of garlic powder in the preparation of kochujang was determined from the result of a sensory evaluation test. Garlic-added kochujang showed the highest points when the added content was 3%. Fermentation of kochujang led to the decreased leptin secretion and increased glycerol release in the 3T3-L1 adipocytes. The addition of garlic to the kochujang also resulted in reduced leptin secretion and induced lipolysis. Since these results suggested that addition of garlic into kochujang can improve the anti-adipogenic effects of kochujang, it might be possible to develop garlic-added kochujang as an antiobesity-functional kochujang.

• Journal of Veterinary Science
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• v.22 no.4
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• pp.55.1-55.17
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• 2021

Background: Naringenin and its glycoside naringin are well known citrus flavonoids with several therapeutic benefits. Although the anti-adipogenic effects of naringenin and naringin have been reported previously, the detailed mechanism underlying their anti-adipogenesis effects is poorly understood. Objectives: This study examined the anti-adipogenic effects of naringenin and naringin by determining differential gene expression patterns in these flavonoids-treated 3T3-L1 adipocytes. Methods: Lipid accumulation and triglyceride (TG) content were determined by Oil red O staining and TG assay. Glucose uptake was measured using a 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose fluorescent d-glucose analog. The phosphorylation levels of AMP-activated protein kinase (AMPK) and acetyl Co-A carboxylase (ACC) were observed via Western blot analysis. Differential gene expressions in 3T3-L1 adipocytes were evaluated via RNA sequencing analysis. Results: Naringenin and naringin inhibited both lipid accumulation and TG content, increased phosphorylation levels of both AMPK and ACC and decreased the expression level of 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) in 3T3-L1 adipocytes. RNA sequencing analysis revealed that 32 up-regulated (> 2-fold) and 17 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Scd1, Mogat1, Dgat, Lipin1, Cpt1a, and Lepr, were normalized to the control level in naringenin-treated adipocytes. In addition, 25 up-regulated (> 2-fold) and 25 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Fabp5, Scd1, Srebf1, Hmgcs1, Cpt1c, Lepr, and Lrp1, were normalized to the control level by naringin. Conclusions: The results indicate that naringenin and naringin have anti-adipogenic potentials that are achieved by normalizing the expression levels of lipid metabolism-related genes that were perturbed in differentiated 3T3-L1 cells.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.4.1-4.15
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• 2022

Background: Flavonoids are natural polyphenols found widely in citrus fruit and peel that possess anti-adipogenic effects. On the other hand, the detailed mechanisms for the antiadipogenic effects of flavonoids are unclear. Objectives: The present study observed the anti-adipogenic effects of five major citrus flavonoids, including hesperidin (HES), narirutin (NAR), nobiletin (NOB), sinensetin (SIN), and tangeretin (TAN), on AMP-activated protein kinase (AMPK) activation in palmitate (PA)-treated HepG2 cells. Methods: The intracellular lipid accumulation and triglyceride (TG) contents were quantified by Oil-red O staining and TG assay, respectively. The glucose uptake was assessed using 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) assay. The levels of AMPK, acetyl-CoA carboxylase (ACC), and glycogen synthase kinase 3 beta (GSK3β) phosphorylation, and levels of sterol regulatory element-binding protein 2 (SREBP-2) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) expression were analyzed by Western blot analysis. The potential interaction between the flavonoids and the γ-subunit of AMPK was investigated by molecular docking analysis. Results: The flavonoid treatment reduced both intracellular lipid accumulation and TG content in PA-treated HepG2 cells significantly. In addition, the flavonoids showed increased 2-NBDG uptake in an insulin-independent manner in PA-treated HepG2 cells. The flavonoids increased the AMPK, ACC, and GSK3β phosphorylation levels and decreased the SREBP-2 and HMGCR expression levels in PA-treated HepG2 cells. Molecular docking analysis showed that the flavonoids bind to the CBS domains in the regulatory γ-subunit of AMPK with high binding affinities and could serve as potential AMPK activators. Conclusion: The overall results suggest that the anti-adipogenic effect of flavonoids on PA-treated HepG2 cells results from the activation of AMPK by flavonoids.

• Journal of Life Science
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• v.29 no.2
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• pp.209-214
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• 2019

Sargassum horneri (Turner) C. Agardh is a marine brown algae widely distributed in the North Pacific Ocean. It is known for its anti-inflammatory and anti-atopic effects. In this study, we determined the effects of ethanol extract of Sargassum horneri (Turner) C. Agardh (EESH) on anti-obesity activities in 3T3-L1 preadipocytes. Our results indicated that treatment with EESH decreased the 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. The concentrations of cellular triglycerides were also reduced in 3T3-L1 cells after treatment with EESH. Triglyceride content was inhibited by 13%, 16%, and 23% after treatment with 250, 500, and $1,000{\mu}g/ml$ of EESH in 3T3-L1 cells, respectively. Western blotting analysis showed that EESH suppressed adipogenic transcription factor expression in a dose dependent manner. Specifically, it suppressed cytidine-cytidine-adinosine-adenosine-thymidine (CCAAT) /enhancer binding proteins $(C/EBP){\alpha}$, $C/EBP{\beta}$ and peroxisome proliferator-activated receptor $(PPAR){\gamma}$. This indicated that EESH could control the expression of adipogenic transcription factors and inhibit the differentiation of adipocytes. Taken together, these findings demonstrated that EESH showed anti-obesity effects and could have potential uses in the field of nutraceuticals.

• Preventive Nutrition and Food Science
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• v.11 no.4
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• pp.285-288
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• 2006

The anti-adipogenic effect of dongchimi nano juice prepared using a nano-filtering process was investigated by measuring leptin and glycerol levels and the expression of a peroxisome proliferator-activated $receptor-\gamma\;(PPAR\gamma)$ gene as indicators of lipid accumulation or lipolysis. Red pepper powder, seeds of red pepper, garlic, and ginger were added in the preparation of dongchimi. Dongchimi was fermented to reach the optimal fermentation period, followed by nano-filtration in the range of $0.0005\sim0.1\;{\mu}m$. The lactic acid bacteria of dongchimi nano juice were removed completely by a nano-filtering process. Treatment of dongchimi nano juice induced glycerol release in the 3T3-L1 adipocytes and decreased the mRNA expression level of $PPAR\gamma$. These results suggested that dongchimi nano juice may enhance lipolysis and modulate adipogenesis in 3T3-L1 cells.

• Journal of Life Science
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• v.24 no.2
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• pp.137-147
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• 2014

Cortex ulmi pumilae, the cortex of Ulmus davidiana var. japonica, has been used in traditional folk medicine for its anti-inflammatory effect. Although its various bioactivities such as anti-inflammatory, anti-microbial, and anti-cancer, have been reported, the anti-adipogenic activity of cortex ulmi pumilae remains unclarified. In the present study, we investigated the effect of cortex ulmi pumilae extract on adipocyte differentiation in 3T3-L1 preadipocytes. Treatment with cortex ulmi pumilae extract significantly reduced the formation of lipid droplets and triglyceride content in a dose-dependent manner; this is associated with an inhibition of the adipogenic transcription factors, CCAAT/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$). In addition, cortex ulmi pumilae extract treatment during the early stage of adipogenesis showed more efficient anti-adipogenic activity than treatment during other stages of adipogenesis. Cortex ulmi pumilae extract also inhibited cell proliferation and induced G1 arrest of 3T3-L1 cells in the early stage of adipogenesis. This was associated with upregulated expression of Cdk inhibitor p21 and downregulated expression of cyclin E and phospho-Rb, indicating that cortex ulmi pumilae extract blocks mitotic clonal expansion by cell cycle regulation. Taken together, these results suggest that cortex ulmi pumilae extract possesses anti-adipogenic activity through the inhibition of adipocyte differentiation by blocking mitotic clonal expansion.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.5
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• pp.831-836
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• 2010

In addition to spice, black pepper (Piper nigrum Linne : PnL) has been used as herbal medicine because of its function in anti-oxidation, anti-inflammation, and anti-carcinogenesis. Recently, it has been reported that piperine, a component of PnL, inhibits adipocyte differentiation by repressing various adipogenic gene expressions. In this study, we determined whether piperine is a major constituent of PnL that confers the anti-adipogenic activity at whole genome level. Differentiation of 3T3-L1 pre-adipocytes was induced in presence of PnL extract or piperine. To compare genes that are regulated by PnL extract or piperine, we performed expression profiling using microarrays (Agilent Mouse 44k 4plex). RNA samples were labeled with Cy3 and Cy5, respectively. Labeled samples were hybridized to the microarrays. Results were filtered and cut off set p<0.05. Genes exhibiting significant differences in expression level were classified into Gene Ontology (GO)-based functional categories (http://www.geneontology.org) and KEGG (http://www.genome.jp/kegg/). Extract of PnL and its component piperine reduced lipid accumulation in 3T3-L1 cells during adipogenesis. Such anti-adipogenic activity appears to result from down-regulation of transcription factor genes involved in adipogenesis, and other genes involved in fatty acid synthesis, transport, triglyceride synthesis, and carbohydrate metabolism. These genome-wide studies lead to conclude that piperine, as a critical component of PnL, plays common role with PnL in anti-adipogenesis.

• Preventive Nutrition and Food Science
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• v.20 no.1
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• pp.29-37
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• 2015

Most of the wheat germ in cereal grains is removed during the milling process. Various physiological effects have been reported for bioactive substances in wheat germ such as phenolic acids and flavonoids. In this study, the antioxidant and anti-adipogenic effects of ethanol extracts from wheat germ (WGE) and wheat germ fermented with Aspergillus oryzae (F-WGE) were investigated in HepG2 and 3T3-L1 cells. The anti-oxidant activity of F-WGE was demonstrated by a dose-dependent increase in the enhanced scavenging capacity of hydroxyl radicals and $Cu^{2+}$-chelating activity compared to WGE. WGE and F-WGE treatment at doses between 10 and $400{\mu}g/mL$ did not affect the viability of HepG2 and 3T3-L1 cells. Intracellular ROS levels from $Cu^{2+}$-induced oxidative stress were significantly decreased by F-WGE treatment in HepG2 cells compared to WGE. Lipid accumulation was increased in 3T3-L1 adipocytes by $100{\mu}M$ $Fe^{2+}$ treatment, but the accumulation was strongly inhibited by $100{\mu}g/mL$ of WGE and F-WGE treatment. These results suggest that changes in bioactive substances during the fermentation of wheat germ can potentiate scavenging activities against transition metal-induced oxidative stress and lipid accumulation in 3T3-L1 adipocytes. Therefore, we propose that F-WGE is a novel food materials and provided scientific evidences for its efficacy in the development of functional foods.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.4
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• pp.401-408
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• 2017

Salvia plebeia R. Br. (Lamiaceae) has been used in folk medicines in Asian countries, including Korea and China, to treat inflammatory diseases. The focus of our research was on the anti-adipogenic activity of ethanol extract from Salvia plebeia R. Br. (SPE) in 3T3-L1 adipocytes. This study investigated inhibition of differentiation and lipogenesis upon SPE treatment in 3T3-L1 cells. The results reveal that SPE at non-cytotoxic concentration significantly suppressed triglyceride accumulation and reduced expression of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein-alpha, and sterol regulatory element-binding protein as adipogenic transcription factors in 3T3-L1 adipocytes compared to non-treated control cells. Inducible phosphorylation of AMP-activated protein kinase, acetyl CoA carboxylase, and hormone-sensitive lipase as well as carnitine palmitoyltransferase-1 mRNA expression increased upon SPE treatment, which suppressed expression of fatty acid synthase. In conclusion, these results demonstrate that SPE can inhibit expression of adipogenic genes in 3T3-L1 adipocytes. Our study suggests that SPE has potential anti-obesity effects and is a novel therapeutic functional agent with anti-adipogenic activity via reduction of lipogenesis.

• Journal of Korean Medicine for Obesity Research
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• v.20 no.2
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• pp.109-121
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• 2020

Objectives: Tanshinone I is a bioactive constituent in Salvia miltiorrhiza. At present, the anti-obesity effect and mechanism of tanshinone I are not fully understood. Here we investigated the effect of tanshinone I on lipid accumulation in 3T3-L1 preadipocytes and zebrafish. Methods: Lipid accumulation and triglyceride (TG) content in 3T3-L1 cells were determined by Oil Red O staining and AdipoRed assay, respectively. The expression and phosphorylation levels of adipogenic/lipogenic proteins in 3T3-L1 cells were evaluated by Western blotting. The messenger RNA (mRNA) expression levels of adipogenic/lipogenic markers and leptin in 3T3-L1 cells were measured by reverse transcription polymerase chain reaction (RT-PCR). Lipid accumulation in zebrafish was assessed by LipidGreen2 staining. Results: Tanshinone I at 5 μM largely blocked lipid accumulation and reduced TG content in differentiating 3T3-L1 cells. Furthermore, tanshinone I decreased the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and perilipin A but also the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. In addition, tanshinone I increased the phosphorylation of adenosine 3',5'-cyclic monophosphate (cAMP)-activated protein kinase (AMPK) while decreased the intracellular adenosine triphosphate (ATP) content with no change in the phosphorylation and expression of liver kinase-B1 in differentiating 3T3-L1 cells. Importantly, tanshinone I also reduced the extent of lipid deposit formation in developing zebrafish. Conclusions: These findings demonstrate that tanshinone I has strong anti-adipogenic effects on 3T3-L1 cells and reduces adiposity in zebrafish, and these anti-adipogenic effect in 3T3-L1 cells are mediated through control of C/EBP-α, PPAR-γ, STAT-3, FAS, ACC, perilipin A, and AMPK.