• Animal Bioscience
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• v.35 no.2
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• pp.155-165
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• 2022

Objective: This study was performed to examine whether the porcine glutamic-oxaloacetic transaminase 1 (GOT1) gene has important functions in regulating adipocyte differentiation. Methods: Porcine GOT1 knockout and overexpression vectors were constructed and transfected into the mouse adipogenic 3T3-L1 cells. Lipid droplets levels were measured after 8 days of differentiation. The mechanisms through which GOT1 participated in lipid deposition were examined by measuring the expression of malate dehydrogenase 1 (MDH1) and malic enzyme (ME1) and the cellular nicotinamide adenine dinucleotide phosphate (NADPH) content. Results: GOT1 knockout significantly decreased lipid deposition in the 3T3-L1 cells (p<0.01), whereas GOT1 overexpression significantly increased lipid accumulation (p<0.01). At the same time, GOT1 knockout significantly decreased the NADPH content and the expression of MDH1 and ME1 in the 3T3-L1 cells. Overexpression of GOT1 significantly increased the NADPH content and the expression of MDH1 and ME1, suggesting that GOT1 regulated adipocyte differentiation by altering the NADPH content. Conclusion: The results preliminarily revealed the effector mechanisms of GOT1 in regulating adipose differentiation. Thus, a theoretical basis is provided for improving the quality of pork and studies on diseases associated with lipid metabolism.

• Journal of Life Science
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• v.29 no.4
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• pp.447-454
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• 2019

Fat accumulation in adipocytes occurs through the process of adipogenesis in which preadipocytes differentiate into adipocytes. Obesity is a metabolic disorder caused by excessive accumulation of fat in the body, which increases the incidence of cardiovascular diseases, hypertension, type 2 diabetes, hyperlipidemia, and various cancers. Recently, inhibition of adipocyte differentiation was shown to be a potential antiobesity strategy. In this study, the inhibitory effect of dichloromethane fractions from Illicium verum Hooker fil. water extract on the differentiation of 3T3-L1 preadipocytes to adipocytes was investigated. Dichloromethane fractions from I. verum Hooker fil. significantly inhibited adipocyte differentiation when applied during the adipocyte differentiation process, as assessed by measuring fat accumulation using Oil-red O staining. In addition, dichloromethane fractions from I. verum Hooker fil. reduced important adipogenic transcription factors, such as CCAAT/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, and peroxisome proliferator activated receptor ${\gamma}$ ($PPAR{\gamma}$). The expression of FAS and LPL, which are terminal differentiation markers of mature adipocytes, was also reduced in the 3T3-L1 adipocytes treated with dichloromethane fractions from I. verum Hooker fil. In addition, the treatment significantly inhibited mitotic clonal expansion, which is essential for adipocyte differentiation, by arresting the G1 phase of the cell cycle. Taken together, these results suggest that dichloromethane fractions from I. verum Hooker fil. may be a natural material with antiobesity effects.

• 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 Physiology & Pathology in Korean Medicine
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• v.33 no.1
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• pp.17-24
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• 2019

Cymbopogon citratus, commonly know as lemongrass, prossesses strong antioxidant, anti-tumor and anti-inflammatory properties. Howerver, its anti-obesity activity remains to be elucidated. This study investigated the effect of ethanol extract of Cymbopogon citratus on adipogenesis, and its underlying mechanism, in 3T3-L1 preadipocytes. The results demonstrated that ethanol extracts of Cymbopogon citratus effectively suppressed intercellular lipid accumulation at non-toxic concentrations, and was associated with the down-regulation of adipocyte-specific transcription factors, including $C/EBP{\alpha}$ and $PPAR{\gamma}$, and phosphorylation of $AMPK{\alpha}$. Furthermore, ethanol extracts of Cymbopogon citratus increased p21 and p21 expression, while the expression of CDK2, cyclin A and cyclin B1 was reduced. As a result, ethanol extracts of Cymbopogon citratus seems to induce G0/G1 cell cycle arrest of 3T3-L1 cells. On the other hand, ERK and Akt signaling pathways were not involved in anti-adipogenesis by ethanol extracts of Cymbopogon citratus. Taken together, theses results suggest that ethanol extracts of Cymbopogon citratus inhibits adipocyte differentiation in 3T3-L1 cells and can be used as a safe and efficient natural substance to manage anti-obesity.

• Journal of Nutrition and Health
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• v.38 no.8
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• pp.649-655
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• 2005

Nutrigenomics refers to research that investigates the interaction between nutrition and the human genome. Caffeine in tea and coffee is widely and routinely consumed by people. This study was performed to confirm the effect of caffeine treatment on the gene expression and cytokine profiling in 3T3-L1 adipocyte cells using microarray and protein array methodology. Treatment of caffeine in 3T3-L1 adipocyte cells increased expression of several genes related with obesity including adipocyte C1Q and collagen domain containing (ACDC), Adipsin (ADN), uncoupling protein 3(UCP3), while glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is known as lipid storage enzyme, was decreased by caffeine treatment. Furthermore, cytokines, such as interleukin-3 (IL-3), interleukin-12(IL-12), interleukin-13 (IL-13), granulocyte colony stimulating factor (GCSF), granulocyte macrophage colony stimulating factor (GM-CSF) and vascular endothelial growth factor (VEGF), were decreased in caffeine treated 3T3-L1 adipocyte cells. These results provided interesting information about the genes related with caffeine and cytokine expression profiling in obesity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.86-92
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• 2007

Hibiscus sabdariffa L., a tropical beverage material, is used commonly as in folk medicine against hypertension, pyrexia, inflammation, liver disorders, and obesity. However, the mechanism by which Hibiscus sabdariffa L. modulates adipogenic differentiation is remained to be elusive. This report was designed to investigate the inhibitory effect of Hibiscus extract on insulin signaling pathway during adipocyte differentiation in 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were differentiated with isobutylmethylxanthine, dexamethasone, and insulin (MDI) and followed by the addition of Hibiscus extract. Treatment with Hibiscus resulted in a decrease of lipid droplet accumulation, which was suppressed by PI-3 kinase inhibitor wortmannin in 3T3-L1 preadipocytes. Also, Hibiscus extract markedly attenuated the mRNA expression of adipogenic transcriptional factor PPAR${\gamma}$ and adipogenic hormon Leptin during adipogenesis. However, it did not affect the expression of adiponectin in 3T3-L1 preadipocytes differentiated with MDI mixture. Furthermore, Adipogenic differentiation by MDI mixture increased the phosphorylation and expression of PI3-Kinase and Akt in 3T3 preadipocytes, which was markedly suppressed by Hibiscus extract treatment. Taken together, our results suggest that Hibiscus extract suppressed the adipogenic differentiation of 3T3 preadipocytes through activation of PI3-Kinase and Akt signaling pathway.

• Toxicological Research
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• v.34 no.1
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• pp.13-21
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• 2018

Anthocyanins are naturally occurring water-soluble polyphenolic pigments in plants that have been shown to protect against cardiovascular diseases, and certain cancers, as well as other chronic human disorders. However, the anti-obesity effects of anthocyanins are not fully understood. In this study, we investigated the effects of anthocyanins isolated from the fruit of Vitis coignetiae Pulliat on the adipogenesis of 3T3-L1 preadipocytes. Our data indicated that anthocyanins attenuated the terminal differentiation of 3T3-L1 preadipocytes, as confirmed by a decrease in the number of lipid droplets, lipid content, and triglyceride production. During this process, anthocyanins effectively enhanced the activation of the AMP-activated protein kinase (AMPK); however, this phenomenon was inhibited by the co-treatment of compound C, an inhibitor of AMPK. Anthocyanins also inhibited the expression of adipogenic transcription factors, including peroxisome proliferator-activated receptor-${\gamma}$, CCAAT/enhancer-binding protein a and b, and sterol regulatory element-binding protein-1c. In addition, anthocyanins were found to potently inhibit the expression of adipocyte-specific genes, including adipocyte fatty acid-binding protein, leptin, and fatty acid synthase. These results indicate that anthocyanins have potent anti-obesity effects due to the inhibition of adipocyte differentiation and adipogenesis, and thus may have applications as a potential source for an anti-obesity functional food agent.

• Journal of Life Science
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• v.24 no.5
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• pp.476-484
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• 2014

Obesity, which is characterized by a state of mild chronic inflammation, is known to cause metabolic diseases. This study was carried out to investigate the effect of lyophilized dropwort vinegar powder (DVP) on adipocyte differentiation and inflammation in T3-L1 preadipocyte and RAW 264.7 macrophage cell lines. DVP inhibited the differentiation of 3T3-L1 preadipocytes induced by a mixture of IBMX, dexamethasone, and insulin (MDI). Western blot analysis of cell lysates showed that DVP decreased the levels of two major transcription factors involved in adipogenesis, peroxisome proliferator- activated receptor-${\gamma}$ (PPAR-${\gamma}$) and CCAAT-enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$). DVP also significantly suppressed lipopolysaccharide (LPS)-induced production of nitric oxide (NO), and this was accompanied by a decrease in inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression. These results demonstrate that DVP inhibits MDI-induced adipocyte differentiation of 3T3-L1 cells and LPS-induced inflammation in RAW 264.7 macrophage cells. The findings indicate that this natural product may be a good candidate as to prevent metabolic diseases.

• The Journal of Korean Medicine
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• v.17 no.2 s.32
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• pp.145-160
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• 1996

Effects of Ryangkeogsanwhatang on the obesity of white mouse induced by gold thioglucose. It is researched to elucidate the effect of Ryangkeogsanwhatang on the obesity of white mouse induced by gold thioglucose and the differentiation and growth of preadipocyte, 3T3-L1. The result were as follows. 1. Ryangkeogsanwhatang extract improved the blood level of transaminase changed by the obesity of white mouse induced by gold thioglucose. 2. Ryangkeogsanwhatang extract inhibited the increase of liver fat and body fat induced by the obesity of white mouse induced by gold thioglucose. 3. Ryangkeogsanwhatang extract inhibited the increase of body weight induced by the obesity of white mouse induced by gold thioglucose. 4. Ryangkeogsanwhatang extract inhibited the growth of undifferentiate preadipocyte 3T3-L1. 5. Ryangkeogsanwhatang extract showed inhibitory on the differentiation of preadipocyte 3T3-L1. The above results suggest that the Ryangkeogsanwhatang extract may be used on the obesity induced by the overgrowth and differentiation of adipocyte, and the accumulation of fat in liver and body.

• The Journal of Pediatrics of Korean Medicine
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• v.27 no.4
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• pp.17-30
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• 2013

Objectives This experimental study was designed to investigate the effects of Mulberry leaves contained herbal mixture (MLHM) on body weight, serum lipid level and adipocyte differentiation in high fat diet-fed obese mice. Methods Four-week old mice (wild-type C57/BL6) were used for all experiments. Cells were incubated with MLHM at the indicated concentration (0.04-4mg/ml) for 24h, and growth rate was assessed by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. 3T3-L1 preadipocytes were incubated in DMEM for 2 days with the indicated concentrations of MLHM, and on Day 6, the cells were fixed and the cellular lipid contents were assessed by Oil-Red-O staining. The expression of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR ${\gamma}$) and cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ (C/EBP ${\alpha}$) as adipocyte-specific proteins were determined by real time RT-PCR and western blotting. In addition, body weight gain and serum lipid levels were measured in the mice with obesity induced by the high fat-diet for four weeks. Results Though MLHM did not show toxicity even at the concentration of 4mg/ml, MLHM significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner. Also, MLHM significantly reduced the expressions of PPAR ${\gamma}$ and C/EBP ${\alpha}$ in a dose-dependent manner. Furthermore, MLHM significantly reduced body weight gain and LDL-cholesterol contents in high fat diet-fed obese mice. Conclusions These results demonstrate that MLHM exerts anti-obesity effect in 3T3-L1 cells and mice with obesity by high-fat diet.