• 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.

• Journal of Korean Medicine for Obesity Research
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• v.20 no.1
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• pp.1-9
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• 2020

Objectives: Artemisinin and its derivatives extracted from Artemisia annua, a Chinese herbal medicine, have variable biological effects due to structural differences. Up to date, the anti-obesity effect of dihydroartemisinin (DHA), a derivative of artemisinin, is unknown. The purpose of this study was to investigate the anti-adipogenic and lipolytic effects of DHA on 3T3-L1 preadipocytes. Methods: Oil Red O staining and AdipoRed assay were used to measure lipid accumulation and triglyceride (TG) content in 3T3-L1 cells, respectively. Cell count analysis was used to determine the cytotoxicity of 3T3-L1 cells. Western blot and real-time reverse transcription polymerase chain reaction analyses were used to analyze the expression of protein and mRNA in 3T3-L1 cells, respectively. Results: DHA at 5 μM markedly inhibited lipid accumulation and reduced TG content in differentiating 3T3-L1 cells with no cytotoxicity. Furthermore, DHA at 5 μM inhibited the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and perilipin A as well as the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. Moreover, while DHA at 5 μM had no effect on the mRNA expression of adiponectin, it strongly suppressed that of leptin in differentiating 3T3-L1 cells. However, DHA at 5 μM had no lipolytic effect on differentiated 3T3-L1 cells, as assessed by no enhancement of glycerol release. Conclusions: These results demonstrate that DHA at 5 μM has a strong anti-adipogenic effect on differentiating 3T3-L1 cells through the reduced expression and phosphorylation of C/EBP-α, PPAR-γ, FAS, perilipin A, and STAT-3.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.46-46
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• 2023

It has been reported that Rosa acicularis has anti-obesity activity by inhibiting the digestive lipase activity. However, there is a lack of clear in vitro studies regarding the anti-obesity activity of Rosa acicularis. Therefore, in this study, we aimed to verify the anti-obesity activity of Rosa acicularis leaves (RAL) and elucidate its mechanism of action in 3T3-L1 preadipocytes. RAL dose-dependently inhibited the accumulation of lipid droplets and triacylglycerol. RAL had no effect on cell proliferation and survival in undifferentiated 3T3-L1 cells, but it inhibited cell proliferation in differentiating 3T3-L1 cells. RAL increased ATGL, p-HSL, and HSL, and decreased perilipin-1 in differentiating 3T3-L1 cells. In addition, RAL reduced lipid droplet accumulation and increased free glycerol content in differentiated 3T3-L1 cells. RAL increased ATGL and HSL in differentiated 3T3-L1 cells. Also, RAL increased p-AMPK, PPARγ, UCP-1, and PGC-1α in differentiating 3T3-L1 cells. AMPK inhibition by Compound C attenuated RAL-mediated increase of ATGL, HSL, PPARγ, and UCP-1 in 3T3-L1 cells. Taken together, it is thought that RAL may inhibit lipid accumulation through lipolysis and thermogenesis via the activation of AMPK in adipocytes.

• Journal of the Korean Society of Food Culture
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• v.33 no.5
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• pp.464-471
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• 2018

Purpose: This study examined the effects of ${\alpha}$-lipoic acid in diluted solvents on cell growth in 3T3-L1 cells according to the treated concentration and times. Methods: Adipocyte 3T3-L1 cell were cultured. Confluent cells underwent starvation with SFM for 1 day and then were cultured in a medium containing various concentrations 0, 100, 200, and $400{\mu}mol/L$ of ${\alpha}$-lipoic acid. The cell viability was measured using the EZ Cytox assay kit. In addition, the effect of ${\alpha}$-lipoic acid of diluted solvents on the cell growth in 3T3-L1cells was examined according to the treated concentration and times. Results: The ${\alpha}$-lipoic acid diluted ethanol inhibited cell proliferation in a dose and time dependent manner. The ${\alpha}$-lipoic acid diluted ethanol induced adipocyte 3T3-L1 cells proliferation with an adipocyte inducer. In addition, ${\alpha}$-lipoic acid inhibited adipocyte 3T3-L1 growth in a dose and time dependent manner (p<0.05). Conclusion: This study showed that a treatment with ${\alpha}$-lipoic acid diluted ethanol inhibits cell growth of, adipocyte 3T3-L1 cells induced with an adipocyte inducer, ($200{\mu}mol/L$ of ${\alpha}$-lipoic acid) treated for 48 hr.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.6
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• pp.541-545
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• 2000

This study examines the effects of alginic acid, a source of dietary fiber, in a glucose-derived media. In particular, we examined how the presence or absence of alginic acid affected the differentiation and triglyceride densities of 3T3-L1 cells. We established that the addition of insulin-like growth factor-I (IGE-I) to 3T3-L1 cells results in acceleration of differentiation. We sought to determine the role of alginic acid in the production of fat by adding alginic acid to 3T3-L1 cells and examining its ability to limit or potentiate this stimulatory effects of IGE-I and IGF binding proteins. We have determined that alginic acid restricts 3T3-L1 cell differentiation and the creation of triglycerides, effectively attenuating 3T3-L1 cell metablolism and growth.

• BMB Reports
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• v.51 no.5
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• pp.207-208
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• 2018

Chitinase-Like Proteins (CLPs) are an evolutionarily conserved protein which lose their enzymatic activity for degrading chitin macromolecules. Chitinase-3-like-1 (Chi3l1) is a type of CLP that is highly expressed in epithelial cells, macrophages, etc., and is known to have correlations with type 2 inflammation and cancer. Although the increased level of Chi3l1 in the blood was reported in various disease patients, the function of Chi3l1 in adaptive immunity has been totally unknown. Recently, we found that Chi3l1 is expressed in T cells and has a negative regulatory role in T-cell activation and proliferation. A genetic ablation study of Chi3l1 in T cells showed hyperresponsiveness to TcR stimulation, which increased proliferation and Th1 differentiation. A significant increase of $IFN{\gamma}$ signaling in Chi3l1-deficient T cells synergistically increased Th1 and CTL functions against melanoma cells in vitro and in vivo. In addition, targeted knockdown by Chi3l1 siRNA complexed with the cell-penetrating peptide dNP2, which showed decreased pulmonary melanoma metastasis with increased infiltration of Th1 and CTL in the lung. This study first suggests that Chi3l1 is a novel regulator of Th1/CTL responses and could be a target for treating cancer to increase tumor immunity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1409-1415
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• 2009

Obesity is especially a serious health problem in industrialized countries, because it is considered to be a risk factor associated with the genesis or development of various metabolic diseases, including cardiovascular disease and type 2 diabetes mellitus. The purpose of this study was to investigate the effects of tanshinone IIA from Salvia miltiorrhiza Bunge on induction of apoptossis and inhibition of adipogenesis in in 3T3-L1 preadipocytes and adipocytes. The results demonstrated that tanshinone IIA decreased cell population growth of 3T3-L1 preadipocytes, assessed with the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and LDH (lactate dehydrogenase) assay. Flow cytometric analysis of 3T3-L1 preadipocytes exposed to tanshinone IIA showed that apoptotic cells increased in a timeand dose-dependent manner. Treatment with tanshinone IIA decreased the number of normal cells and increased the number of apoptotic cells in a dose-dependent manner. The induction of apoptosis in 3T3-L1 preadipocytes by tanshinone IIA was mediated through the activation of caspase-3 and Bax, and then through the cleavage of PARP and the down-regulation of Bcl-2. Moreover, tanshinone IIA significantly decreased the amount of intracellular triglycerides and GPDH (glycerol-3-phosphate dehydrogenase) activity in 3T3-L1 adipocytes. Our results suggest that tanshinone IIA efficiently induces apoptosis and inhibits adipogenesis in 3T3-L1 preadipocytes and adipocytes.

• Journal of Animal Science and Technology
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• v.55 no.1
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• pp.19-23
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• 2013

The current study was carried out to determine the effects of Kohlrabi (Brassica oleracea var. gongylodes) on proliferation and differentiation of pig preadipocytes and $_3T_3-L_1$ cells. Pig preadipocytes were isolated from the backfat of the new-born pigs. Twenty-four hours after seeding, the cells were washed with DMEM/F-12 (designated day 0). To measure the cell proliferation, the cells were treated with 25 ng/ml and 100 ng/ml ethanol extracts of Kohlrabi (peel and flesh) for two days (day 0 ~ 2). To measure differentiation, the cells were treated with Kohlrabi for two days (day 0 ~ 2) and cell differentiation was measured on day 6. Twenty-five ng/ml and 100 ng/ml of Kohlrabi peel decreased proliferation of pig preadipocytes by 4.59% and 17.7%, respectively, compared with the control and Kohlrabi flesh by 11.4% and 19.2%, respectively. However, Kohlrabi did not inhibit cell differentiation. To measure the effects of Kohlrabi on proliferation and differentiation of $_3T_3-L_1$ cells, the cells were treated with Kohlrabi for two days in culture, like pig preadipocytes. Kohlrabi (both peel and flesh) did not show any effects on cell proliferation and differentiation. In summary, the results of the current study showed that Kohlrabi decreased proliferation of pig preadipocytes, but no inhibitory effects on differentiation of the cells. Kohlrabi had no effects on proliferation and differentiation of $_3T_3-L_1$ cells.

• The Journal of Korean Medicine
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• v.20 no.3 s.39
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• pp.105-114
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• 1999

To investigate the anti-complementary and cytotoxic effects of oriental prescription, Kuseonwangdogo, on the proliferation of preadipocyte 3T3- L1 cells, we examined biological effects of Kuseonwangdogo. The results obtained were as follows. 1. After 14 days, the body weight of rats treated with Kuseonwangdogo decreased more than that in the control group (p<0.05). However, the weights of liver, spleen and kidney were unchanged. In serum biochemical test, we examined the level of glucose (GLU) and glutamic pyruvic transaminase (GPT). The levels of GOT and CHOL in serum were decreased remarkably by the administration of Kuseonwangdogo (p<0.05). The haematological examination of the tested group showed significant increment of white blood cells (WBC), hemoglobin concentration (HGB), mean corpuscular hemoglobin (MCH) and monocyte (MO). 2. The effect of Kuseonwangdogo on the proliferation of 3T3-L1 cells was tested by the sulforhodamin B(SRB) assay. The high concentration ($100{\mu}l\;and\;200{\mu}l$) of extracts inhibited the proliferation of 3T3- L1 cells. The p-value was <0.01, respectively. 3. The extract of Kuseonwangdogo showed a potent anti -complementary activity. It was suggested that the active principle may be a kind of polysaccharide molecule. 4. The cytotoxic effects of Kuseonwang dogo and its composing herbs in human liver cells (WRL68) and monkey kidney cells (Vero) were examined by the SRB and 3- (4,5- Dimethylthiazol-2-yl) -2,5 diphenyl-2H- tetrazolium bromide (MTT) assay. Cytotoxic effects were not observed.

• Nutrition Research and Practice
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• v.8 no.1
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• pp.33-39
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• 2014

Obesity occurs when a person's calorie intake exceeds the amount of energy burns, which may lead to pathologic growth of adipocytes and the accumulation of fat in the tissues. In this study, the effect and mechanism of pear pomace extracts on 3T3-L1 adipocyte differentiation and apoptosis of mature adipocytes were investigated. The effects of pear pomace extract on cell viability and the anti-adipogenic and proapoptotic effects were investigated via MTT assay, Oil red O staining, western blot analysis and apoptosis assay. 3T3-L1 preadipocytes were stimulated with DMEM containing 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), $5{\mu}g/ml$ insulin and $1{\mu}M$ dexamethasone for differentiation to adipocytes. 3T3-L1 cells were cultured with PBS or water extract of pear pomace. Water extract of pear pomace effectively inhibited lipid accumulations and expressions of PPAR-${\gamma}$ and $C/EBP{\alpha}$ in 3T3-L1 cells. It also increased expression of p-AMPK and decreased the expression of SREBP-1c and FAS in 3T3-L1 cells. The induction of apoptosis was observed in 3T3-L1 cells treated with pear pomace. These results indicate that pear pomace water extract inhibits adipogenesis and induces apoptosis of adipocytes and thus can be used as a potential therapeutic substance as part of prevention or treatment strategy for obesity.