• Asian-Australasian Journal of Animal Sciences
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• v.29 no.3
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• pp.419-427
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• 2016

There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at $42^{\circ}C$ for one hour and then allowed to recover at normal incubation temperature of $37^{\circ}C$ for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to $400{\mu}g/mL$) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

• KSBB Journal
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• v.24 no.6
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• pp.549-555
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• 2009

4 kinds of Regional Special Natural Products (RSNPs), such as Anthrisci radix, Psoraleae semen, Siegesbeckiae herba and Corni fructus were examined to verify for anti-obesity effect. $PPAR\gamma$ (peroxisome proliferator-activated receptor $\gamma$) from 3T3-L1 cell concerning adipocyte differentiation was suppressed by different concentraton of 4 RSNPs with western blot, when treated RSNPs' extract and MDI (IBMX, Dexamethasone, Insulin) at the same time. Also, SREBP-1 (Sterol regulatory element binding protein) controlling lipogenesis and $PPAR\gamma$ expression levels were reduced by these 4 RSNPs' extract, when these chemicals after differentiation of 3T3-L1 cell. And lipid droplets were reduced by 7.5%, 14.4%, 18.3% and 30% at different concentration of Anthrisci radix from Oil Red O staining. Also, it was reduced by 2%, 4.9%, 9.3% and 38% at different concentration of Psoraleae semen. For Siegesbeckiae herba, it was inhibited by 1.4%, 6.4%, 16.4% and 30.1%, respectively. And Corni fructus was also showed by 0.9%, 6.3%, 13.7% and 33% at same concentration of Siegesbeckiae herba. These 4 kinds of RSNPs were expected for a useful material for anti-obesity materials.

• Journal of Applied Biological Chemistry
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• v.61 no.1
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• pp.1-8
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• 2018

The objective of this study was to compare the antihyperlipidemic effects of different Dendropanax morbifera leaf extracts in vitro. The extracts differed in terms of specimen age, harvesting season, and extraction method. RAW 264.7 cells were pretreated with these extracts and stimulated by oxidized low-density lipoprotein. Ethanol was used to induce toxicity in HepG2 cells. Cellular lipid accumulation was quantified using oil red O staining in both these cells. The extracts were evaluated for their inhibitory effects on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. RAW 264.7 cells treated with the 60% ethanol extract of an 8-year-old specimen harvested in November exhibited the lowest lipid accumulation. The 30% ethanol extract of a 5-year-old specimen harvested in May exhibited the greatest protection from ethanol-induced cytotoxicity in HepG2 cells. The hot water extract of an 8-year-old specimen harvested in May showed the greatest inhibition of HMG-CoA reductase. These results showed that D. morbifera extracts prepared from leaves that are harvested in May possess the highest antihyperlipidemic effects.

• Microbiology and Biotechnology Letters
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• v.44 no.1
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• pp.89-97
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• 2016

Obesity is caused by excess accumulation of body fat and contributes to various pathological disorders such as diabetes, hypertension, cardiovascular disease, and cancer. In this study, we investigated the effect of a 30% ethanol extract of Fructus Rosae laevigata (RLE) on adipogenesis in 3T3-L1 adipocytes, measured by triglyceride accumulation and expression of adipogenesis-related transcription factors during differentiation of pre-adipocytes into adipocytes. RLE decreased the intracellular triglyceride contents (assessed by Oil Red-O staining) in a dose-dependent manner. It also downregulated the expression of adipogenic transcription factors and inhibited cell proliferation during the mitotic clonal expansion phase of adipocyte differentiation by inducing G1 phase arrest. We investigated the alterations in the levels of G1 phase arrest-related proteins. The expression of p21 protein significantly increased, while the levels of Cyclin E, Cdk2, and phospho-Rb decreased in a dose-dependent manner in 3T3-L1 cells treated with RLE. These results suggest that RLE inhibits the differentiation of 3T3-L1 adipocytes by suppressing the expression of adipogenic transcription factors and inducing G1 phase arrest in the early stages of adipocyte differentiation.

• Journal of Life Science
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• v.22 no.1
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• pp.49-54
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• 2012

The effects of genistein on cell proliferation and adipogenesis were examined in mouse 3T3-L1 preadipocyte cells. Genistein decreased viability of 3T3-L1 pre-adipocytes in a dose-dependent manner. Oil Red O staining of these cells also indicated that adipogenesis was inhibited by 50 ${\mu}M$ genistein treatment. We investigated the molecular mechanisms involved in the decrease in cell viability in genistein-treated 3T3-L1 cells by conducting an oligo DNA microarray analysis. We selected the sirtuin-1 gene, one of the upregulated genes, for further experimentation because sirtuin-1 belongs to the sirtuin family, which is associated with anti-obesity and anti-inflammation activities. We found that four phytochemicals (resveratrol, capsaicin, daidzein, and genistein) could increase sirtuin-1 expression. Genistein was the strongest inducer of sirtuin-1 among the tested phytochemicals. The inhibition of adipogenesis by genistein was recovered by surtuin-1 siRNA transfection. Overall, these results may further our understanding of the molecular mechanisms underlying the inhibition of proliferation and adipogenesis by genistein in mouse 3T3-L1 cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1126-1131
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• 2010

The antiobesity effect of soymilks fermented with Bacillus subtilis KC-3 (KCCM 42923) from cheonggukjang was compared with other sources of B. subtilis KCCM 11316 and B. subtilis MYCO. The antiobesity effect was investigated by measuring the release of leptin, Oil red O staining, glycerol secretions and adipogenic transcription factor by reverse transcription-polymerase chain reaction (RT-PCR) in the 3T3-L1 adipocytes. Fermented soymilk with B. subtilis KC-3 (F-KC) led to decrease levels of leptin secretion and increase levels of glycerol secretion in the cells. In addition, F-KC reduced contents of Oil red O dye in the 3T3-L1 adipocytes. Also, mRNA expression levels of both SREBP-1c (sterol regulatory element-binding protein 1-c) and PPAR-$\gamma$ (peroxisome proliferator-activated receptor-$\gamma$), which are adipogenic transcription factor, in cells treated with F-KC were markedly down regulated. These results demonstrate that the Bacillus subtillis fermented soymilk (F-KC) decreased lipid content in 3T3-L1 adipocytes by inhibiting lipogenesis. All B. subtilis fermented soymilks had shown antiobesity activities, however, F-KC exhibited the strongest antiobesity effect in the 3T3-L1 adipocytes. Our study suggests that especially F-KC increased the potential of antiobesity effects.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.2
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• pp.200-206
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• 2014

We investigated the in vitro and in vivo anti-obesity effects of extracts from young Akebia quinata D. leaves, including hot water (AQH) and 80% ethanol (AQE) extracts. The inhibitory effects of AQH and AQE on lipid accumulation in 3T3-L1 cells were examined by Oil Red O staining. Compared to control, lipid accumulation was significantly reduced by 18.3% with the treatment upon AQE at a concentration of $5{\mu}g/mL$. The levels of intracellular triglycerides and free glycerol were also reduced by 52.8% and 9.1% at the same concentration of AQE. The in vivo anti-obesity effect of AQE was evaluated in terms of body and white adipose tissue weights in ICR mice. Experimental groups were divided into the following five groups: normal diet (ND), high fat diet (HFD), high fat diet with 60 mg/kg/day of Orlistat (HFD-RF), high fat diet with 200 mg/kg/day of AQE (HFD-AL), and high fat diet with 600 mg/kg/day of AQE (HFD-AH). Feeding of HFD for eight weeks resulted in significant increases in body weight as well as weight gain compared to the ND group. HFD-AH group showed reduced body weight, weight gain, epididymal white adipose tissue weight, and perirenal white adipose weight as compared to the HFD group. These results indicate that AQE supplementation might have beneficial effects on anti-obesity by inhibiting lipid accumulation.

• Journal of Korean Medicine for Obesity Research
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• v.19 no.2
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• pp.106-112
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• 2019

Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4^th day and insulin medium at 6^th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×10⁷ CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

• Journal of Life Science
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• v.25 no.7
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• pp.773-779
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• 2015

This study examined extracts from five different kinds of lees and nuruk and their organic solvent fractions in terms of several biological functions, such as anti-adipogenic, anti-inflammatory, and anti-proliferative activities. The anti-adipogenic activity was investigated by treating mouse pre-adipocyte 3T3-L1 cells with one extract (YE) and four organic solvent fractions (YAc, PAc, RAc, and WPAc) during adipogenesis. Among the treated samples, the ethyl acetate fraction of W-Ju lees (WPAc) showed the strongest anti-adipogenic effect, which was confirmed with oil red O staining and down-regulation of pro-adipogenic genes such as PPAR-gamma and SCD-1. Treatment with WPAc also reduced the expression of PPAR-gamma in a time-dependent manner. The effects of five different extracts were examined on nitric oxide (NO) production in mouse RAW 264.7 cells to determine anti-inflammatory activity. The ethyl acetate fraction of B-Ju lees (PAc) significantly decreased NO production in LPS-stimulated RAW 264.7 cells and it also inhibited NO production in a dose-dependent manner. The PAc fraction also dramatically decreased the viability of human colorectal cancer HCT116 cells in a dose-dependent manner. In addition, PAc increased the expression of NAG-1 and ATF3 genes in a dose dependent manner. Overall, these results indicate that lees and nuruk have several biological functions, including anti-adipogenic, anti-inflammatory, and anti-proliferative activities.

• Korean Journal of Food Science and Technology
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• v.45 no.1
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• pp.90-96
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• 2013

The anti-obesity effect of ethanol xtract and their fractions from Rumex Crispus L. on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil red O assay, western blot and real-time PCR analysis. Ethyl acetate fraction of Rumex crispus L. significantly inhibited adipocyte differentiation when treated during the adipocyte differentiation process, as assessed by measuring fat accumulation using Oil red O staining. In inducing differentiation of 3T3-L1 preadipocytes in the presence of an adipogenic cocktail, isobutylmethylxanthine (IBMX), dexamethasone- and insulin-along with ethyl acetate fraction residue processing treatment significantly decreased protein expression of obesity-related proteins, such as peroxisome-proliferators-activated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). These results indicate that ethyl acetate fraction of Rumex crispus L. is the most effective candidate for preventing obesity. However further studies will be needed to identify the active compounds that confer the anti-obesity activity of ethyl acetate fraction from Rumex crispus L.