• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.8
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• pp.1199-1204
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• 2003

The aim of this study was to investigate the role of phosphatidylinositol 3-kinase (PI3 kinase) and the mitogenactivating protein (MAP) kinase pathway on the differentiation of ovine preadipocytes. In order to investigate this issue, we monitored glycerol 3-phosphate dehydrogenase (GPDH) activity during differentiation with specific inhibitors of PI3 kinase and MAP kinase-Erk kinase, LY294002 and PD098059, respectively. The preadipocytes, which were obtained from ovine subcutaneous adipose tissues, were proliferated to confluence and then differentiated to adipocytes in differentiation medium with each inhibitor for 10 days. The confluent preadipocytes and differentiated adipocytes at days 3, 7 and 10 were harvested for assay of GPDH activity. LY294002 inhibited the differentiation program in dose- and day-dependent manners during 10 days of differentiation. PD098059 did not affect GPDH activity during differentiation. Furthermore, the expression of peroxisome proliferator-activated receptor ${\gamma}2$ (PPAR-${\gamma}2$), the representative early gene of differentiation, was markedly reduced by LY294002 treatment, although PD098059 did not change it. Our results demonstrated that the activation of PI3 kinase contributes to the differentiation process of ovine preadipocytes.

• Journal of Animal Science and Technology
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• v.54 no.2
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• pp.103-109
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• 2012

Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a $0.4{\mu}m$ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.

• Journal of Sasang Constitutional Medicine
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• v.24 no.4
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• pp.75-83
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• 2012

Objectives : Although Yeoldahanso-tang (YDHST) has been widely used for treatment of obesity and its related diseases such as hyperinsulinemia and hypertension for Taeeumin, no scientific evidence has reported yet to support its ability to work against these metabolic disorders. Our study was aimed to investigate the anti-obesity effect of YDHST extract on the cellular differentiation of 3T3-L1 preadipocytes into adipocytes. Methods : 3T3-L1 preadipocytes were differentiated into adipocytes by adding insulin, dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) for 8 days in the absence or presence of YDHST extract. Anti-obesity effects of YDHST extract were evaluated by Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, triglyceride contents, and leptin production. Results : YDHST extract remarkably prevented lipid accumulation with no cytotoxicity in the differentiated 3T3-L1 cells. In addition, YDHST extract decreased contents of triglyceride 3T3-L1 adipocytes. Consistently, YDHST extract caused a significant inhibition of GPDH activity and leptin production in a dose-dependent manner. Conclusions : Our findings suggest that Sasang constitutional herbal formula YDHST for Tae-eumin has anti-obesity activity by regulation of the adipogenesis process in vitro. Additional study will be required to further confirm the inhibitory effect on adipocyte differentiation by using in vivo animal model.

• Nutrition Research and Practice
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• v.9 no.4
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• pp.439-444
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• 2015

BACKGROUND/OBJECTIVES: This study was conducted to investigate the effects of fermented soybean (FS) extract on adipocyte differentiation and fat accumulation using cultured 3T3-L1 adipocytes. MATERIALS/METHODS: 3T3-L1 adipocytes were treated with FS and nonfermented soybean (NFS) extract during differentiation for 10 days in vitro. Oil red O staining was performed and glycerol-3-phosphate dehydrogenase (GPDH) activity was measured for analysis of fat accumulation. Expressions of adipogenic genes were measured. RESULTS: Soluble extract of soybean fermented with Aspergillus oryzae GB107 contained higher levels of low-molecular-weight protein than conventional soybean protein did. FS extract ($50{\mu}g/ml$) inhibited adipocyte differentiation and fat accumulation during differentiation of 3T3-L1 preadipocytes for 10 days in vitro. Significantly lower GPDH activity was observed in differentiated adipocytes treated with the FS extract than those treated with NFS extract. Treatment with FS extract resulted in decreased expression levels of leptin, adiponectin, and adipogenin genes, which are associated with adipogenesis. CONCLUSIONS: This report is the first to demonstrate that the water-soluble extract from FS inhibits fat accumulation and lipid storage in 3T3-L1 adipocytes. Thus, the soybean extract fermented with A. oryzae GB107 could be used to control lipid accumulation in adipocytes.

• Journal of Sasang Constitutional Medicine
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• v.25 no.1
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• pp.51-61
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• 2013

Objectives We performed the simultaneous analysis for three compounds in Cheongsimyeonja-tang (CSYJT) and evaluated its anti-obesity effect. Methods The column for separation of three compounds was used Gemini $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 275 nm. The injection volume was $10{\mu}L$. 3T3-L1 preadipocytes were differentiated into adipocytes by adding insulin, dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) for 8 days in the absence or presence of CSYJT. Anti-obesity effects of CSYJT were evaluated by Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, triglyceride contents, and leptin production. Results Calibration curves were acquired with $r^2$ >0.9999. The contents of baicalin, wogonoside and baicalein in CSYJT were 14.54-14.65 mg/g, 5.24-5.27 mg/g and 0.01-0.02 mg/g, respectively. CSYJT showed inhibitory effect on lipid accumulation and GPDH activity in the differentiated 3T3-L1 cells. Furthermore, CSYJT significantly decreased contents of triglyceride and leptin production in 3T3-L1 adipocytes. Conclusions These results will be helpful to improve quality control and anti-obesity effect of Taeeumin CSYJT.

• Journal of agriculture & life science
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• v.45 no.2
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• pp.85-92
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• 2011

The current study was conducted to investigate effects of sera taken from Hanwoo at different age on adipocyte differentiation. Sera were taken from Korean native (Hanwoo) steers at 6, 12, 18, and 24 months of age, respectively and supplemented to 3T3-L1 preadipocytes after the cells reached confluence and maintained 10 days thereafter. For the first 2 days (48 h), cells were induced to differentiate by addition of differentiation factors, methylisobutylxanthine, daxamethasone, and insulin. After the differentiation, the cells were incubated without differentiation factors except insulin. The cells lost their fibroblastic shape and showed round-up appearances after 10 days incubation with FBS and the sera of Hanwoo steers. Big lipid droplets appeared in the cells cultured with FBS and the sera taken from Hanwoo at 18 and 24 months of age. After 18 months of age, GPDH activity was statistically higher than 6 and/or 12 months of age (P < 0.05). Based on morphology and Sn-glycerol-3-phosphate dehydrogenase activities, Hanwoo steers expressed aged-dependent adipogenic activities, indicating that aged sera may result in high adipocyte differentiation. It is concluded that the 18 months of age may be 'threshold' to express major adipogenic activities. This may strongly support previous field studies reporting considerable increase in fat contents of Hanwoo carcass at over 18 months of age.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1378-1382
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• 2004

Zinc (Zn) is a micromineral and functions as a cofactor of many enzymes and its deficiency induces retardation of growth and dysfunction of the immune system in animals. This study was conducted to determine lipogenic activity of Zn in bovine intramuscular adipocytes. Preadipocytes were isolated from intramuscular fat depots of 26 month old Korean (Hanwoo) steers and cultured in media containing Zn. At confluence, the cells were treated with insulin, dexamethasone, and 1-methyl-3-isobutyl-xanthine to induce differentiation (accumulation of lipid droplets in cells). The sources of Zn were zinc chloride (${ZnCl}_2$) and zinc sulfate (${ZnSO}_4$), and the final concentrations of both Zn sources were 0, 5, 25, 50 and 100 ${\mu}$M. Glycerol-3-phosphate dehydrogenase (GPDH) activity, an index of adipocyte differentiation, was increased as the concentration of Zn in media increased showing the highest activity (25.74 ng/min/mg protein) at 25 ${\mu}$M of ${ZnSO}_4$. Supplementation of Zn during differentiation of bovine intramuscular adipocytes tended to decrease the production of nitric oxide (NO). Peroxisome proliferator-activated receptor gamma 2(PPAR$\gamma$2) gene expression was increased 10 days after differentiation induction. The current results indicate that Zn has a strong lipogenic activity in cultured bovine intramuscular adipocytes with remarkable suppression of NO production.

• Journal of Pharmacopuncture
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• v.11 no.1
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• pp.149-162
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• 2008

Objectives : The purpose of this study is to investigate the effects of hot pepper extract and capsaicin on the adipogenesis in 3T3-L1 cells, lipolysis in rat epididymal adipocytes and histological changes in porcine adipose tissue. Methods : Inhibiton of preadipocyte differentiation and/or stimulation of lipolysis play important roles in reducing obesity. 3T3-L1 preadipocytes were differentiated with adipogenic reagents by incubating for 3 days in the absence or presence of hot pepper extract or capsaicin ranging from 0.01 to $1mg/m{\ell}$. The effects of hot pepper extract and capsaicin on adipogenesis were examined by measuring GPDH activity and by Oil Red O staining. Mature adipocytes from rat epididymal fat pad was incubated with hot pepper extract or capsaicin ranging from 0.01 to $1mg/m{\ell}$ for 3 hrs. The effects of hot pepper extract and capsaicin on lipolysis were examined by measuring free glycerol released. Fat tissue from pig skin was injected with hot pepper extract or capsaicinCFP ranging from 0.1 to $10mg/m{\ell}$ to examine the effects of hot pepper extract and capsaicin on histological changes under light microscopy. Results : The following results were obtained from present study on adipogenesis of preadipocytes, lipolysis of adipocytes and histological changes in fat tissue. 1. Hot pepper extract and capsaicin inhibited adipogenic differentiation at the concentration of 0.1 and $0.01mg/m{\ell}$, respectively, indicating that capsaicin was more effective in inhibiting adipogenesis than hot pepper extract. 2. Hot pepper extract and capsaicin decreased the activity of glycerol-3-phosphate dehydrogenase(GPDH) at the concentration of 0.1 and $0.01mg/m{\ell}$, respectively, indicating that capsaicin was more effective in inhibiting adipogenic differentiation than hot pepper extract. 3. Hot pepper extract and capsaicin increased glycerol release at the concentration of $0.1mg/m{\ell}$. There was no difference in lipolytic activity between hot pepper extract and capsaicin at the corresponding concentration. 4. Hot pepper extract and capsaicin caused shrinkage of fat cells, resulting in cell death at the concentration of $1.0mg/m{\ell}$, although capsaicin exerted this action over wide area than hot pepper extract. Conclusions : These results suggest that hot pepper extract and capsaicin efficiently inhibited adipogenesis, increased lipolysis of adipocytes and caused to shrink fat cells. Future studies are needed to make use of hot pepper extract pharmacopuncture for the treatment of obesity.

• Archives of Plastic Surgery
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• v.36 no.2
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• pp.135-139
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• 2009

Purpose: The use of an autogenous fat graft has become a common procedure in plastic surgery. However, questions remain concerning on the viability of fat cells and preservation method of aspirated fat. The purpose of this study was to examine the viability of fat cells stored at $-20^{\circ}C$ in the freeze for 1 year after harvest from abdominal liposuction. Methods: Eighteen adults (aged 24 to 65 years old, 16 female and 2 male) were recruited for this study. Harvested aspirated fat tissues were obtained by suction - assisted lipectomy and frozen at $-20^{\circ}C$ commercial refrigerator for one year (average 12.5 months). The viability off at cells in specimens were measured after thawing. The numbers of viable cells were measured on a fluorescence microscope after staining with fluorescein diacetate and propidium iodide. GPDH (Glycerol - 3 - phosphate dehydrogenase) activity was measured. Cell culture was done for 3 weeks. Results: There were no viable cells under the fluorescence microscope, no detectable GPDH activity, and no cultured cells. Conclusion: These findings suggest that aspirated fat after frozen storage for one year at $-20^{\circ}C$ freezer is inadequate to reuse.