• Journal of Korean Medicine for Obesity Research
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• v.22 no.1
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• pp.38-46
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• 2022

Objectives: Juknyeok (JN) is natural liquor extracted from bamboo stems (Phyllostachys bambusoides) and has been used as a traditional Korean medicine for improving vascular function, blood glucose, and treating stroke. Until now, the JN's lipid-lowering effect and underlying mechanism in adipocytes are poorly understood. The aim of this study was to scrutinize the effect of a standardized commercial JN on lipid accumulation during the differentiation of 3T3-L1 preadipocytes. Methods: Lipid and triglyceride (TG) accumulation in differentiating 3T3-L1 preadipocytes were measured by Oil Red O staining and AdipoRed assay, respectively. Cell count analysis was used to ascertain 3T3-L1 cytotoxicity. Immunoblotting and Reverse transcription polymerase chain reaction analysis were used to assess protein and messenger RNA (mRNA) expression levels in 3T3-L1 cells, respectively. Results: Treatment with JN at 25 𝜇l/ml after pH calibration with 6.35 significantly reduced lipid and TG accumulation in differentiating 3T3-L1 preadipocytes without significant cytotoxicity. On mechanistic levels, JN markedly suppressed protein expression levels of CCAAT/enhancer-binding protein (C/EBP)-𝛽 and fatty acid synthase (FAS) during the differentiation of 3T3-L1 preadipocytes. However, JN did not affect the protein expression levels of C/EBP-𝛼, peroxisome proliferator-activated receptor-𝛽/𝛾, and phosphorylation levels of signal transducer and activator of transcription-3/5 in differentiating 3T3-L1 preadipocytes. JN also reduced leptin mRNA expression levels in differentiating 3T3-L1 preadipocytes. Conclusions: JN at 25 𝜇l/ml lowers lipid accumulation and TG content in differentiating 3T3-L1 cells, mediated through the reduced expression levels of C/EBP-𝛽 and FAS.

• The Korea Journal of Herbology
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• v.31 no.4
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• pp.11-18
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• 2016

Objectives : The objective of this study was to investigate the effect of Myrrh 80% ethanol extract on adipocyte differentiation and adipogenesis in 3T3-L1 cell.Methods : Myrrh was prepared by extracting with 80% ethanol. Cell viability was assessed by MTT assay using 3T3-L1 cells. Anti-obesity activity was measured in lipid droplets and triglyceride (TG) accumulation in 3T3-L1 cells. We also analyzed the expression of C/EBPβ, C/EBPα, PPARγ, SREBP1c, and aP2 by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, we observed the production of fatty acid, acetyl-CoA carboxylase and Oil-red O stainingResults : No cytotoxicity from Myrrh 80% ethanol extracts was observed at the concentration of 1, 10, 100 (㎍/㎖) in 3T3-L1 cells. Treatment with Myrrh significantly suppressed the terminal differentiation of 3T3-L1 in a dose-dependent manner, as confirmed by a decrease in triglyceride and Fatty acid and Acetyl-CoA carboxylase. Also, Myrrh exhibited potential adipogenesis inhibition and downregulated the expression of pro-adipogenic transcription factors, such as sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα) and C/EBPβ, and adipocyte expressed genes, such as adipocyte fatty acid binding protein (aP2) and Fas. In addition, lipid accumulation determined by Oil-red O staining showed that Myrrh extract had inhibitory effects on lipid accumulation in 3T3-L1 cells.Conclusions : These results suggest that Myrrh suppresses obesity factors in 3T3-L1 cells. Myrrh may be a useful medical herbs for attenuating metabolic diseases such as obesity.

• The Journal of Korean Medicine
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• v.19 no.1
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• pp.358-367
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• 1998

These studies were conducted to investigate the effects of the various aqua-aqupunctures of Hwanggum(Scutellariae Radix) on the proliferation of 3T3-Ll cells. They were tested by means of Sulforhodamin B(SRB) assay. The results were summerized as follows: All tested aqua-aqupunctures inhibited the proliferation of preadipose 3T3-Ll cells. In case of the dilution of Hwanggum aqua-aqupuncture(HG), the results were quite opposite. In 1000 times dilution of HG(${\times}1000$), a low concentration increased the proliferation of preadipose 3T3-Ll cells, but the high($100{\mu}l\;and\;200{\mu}l$) concentration inhibited it. These results suggest that Hwanggum aqua-aqupunctures may be used on the obesity induced by the overgrowth of preadipose 3T3-Ll cells.

• Food Quality and Culture
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• v.2 no.2
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• pp.85-88
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• 2008

We have investigated the effects of cottonseed extract on the proliferation, differentiation and lipopolysaccharide (LPS)-induced production of local factors in murine clonal osteoblastic MC3T3-E1 cells. Ethanol extract of cotton seed ($4{\sim}63{\mu}g/mL$) significantly increased the proliferatin of MC3T3-E1 cells (p<0.05). Moreover, cottonseed extract ($10{\sim}50{\mu}g/mL$) caused a significant elevation of alkaline phosphatase (ALP) activity and collagen content in the cells. Lipopolysaccharide (LPS) is a potent stimulator of bone resorption in inflammatory diseases. We examined the effect of cottonseed extract on the LPS-induced production of tumor necrosis factor a (TNF-$\alpha$) and nitric oxide (NO) in MC3T3-E1 cells. Treatment with cottonseed extract ($10{\sim}50{\mu}g/mL$) decreased the $5{\mu}g/mL$ LPS-induced production of TNF-$\alpha$ and NO in osteoblasts, suggesting that the antiresorptive action of cottonseed extract may be mediated by decrease in these local factors. This study suggests that cottenseed may contribute to antiresorptive action against osteoblastic cells, resulting in a beneficial effect in promoting the function of osteoblastic cells.

• YAKHAK HOEJI
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• v.38 no.3
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• pp.238-249
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• 1994

The purpose of this research was to investigate effects of baicalin on the differentiation of preadipocytes, 3T3-L1, and to characterize the action of baicalin that affect the responses of 3T3-L1 cells during differentiation. In various culture conditions, effects of baicalin and adrenoreceptor agonists such as phenylephrine(PE) and isoproterenol(IPR) on cell differentiation were examined. Also, effects of the drugs on differentiation, triglyceride(TG) contents, expression of insulin receptor, cAMP contents, the cytosolic $Ca^{2+}$ levels, and amount of calmodulin(CaM) were examined. The results suggest that baicalin has adrenergic receptor blocking activity during the process of differentiation of 3T3-L1 cells and that in the early stage of the adipose conversion, the effect of baicalin on the adipocyte differentiation is mediated by the regulation of insulin receptor expression, but by alterations of the cAMP and the calcium metabolism in the late stage. These results also suggest that the action of baicalin may be significant in the lipid metabolism, lipogenic and lipolytic pathways, of adipose cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.5
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• pp.1315-1320
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• 2003

The purpose of this research was to investigate effects of Bambusae Caulis in Liquamen (BCL) on the synthesis of basement membrane proteins during proliferation and differentiation of 3T3-L1 cells. BCL has been used to relieve the cough and asthma, and remove phlegm in traditional oriental medicines. In recent years. it was studied for its antiinflammatory, antiallergenic. immune-modulating and anticarcinogenic capabilities. We have previously observed that glycyrrhizin stimulates the adipose conversion of 3T3-L1 cells. To investigate effects of BCL on the basement membrane proteins during proliferation and differentiation of 3T3-L1 cells, we have analyzed synthetic amounts of basement membrane components such as type IV collagen and BM40. BCL stimulated the synthesis and secretion of type IV collagen from both 3T3-L1 preadipocytes and adipocytes. The synthesis and secretion of BM40 was not affected by BCL. The continuous addition of BCL markedly stimulated cell growth and increased cell density. These results suggest an important role for type IV collagen in adipocyte differentiation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.5
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• pp.372-379
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• 2004

This study evaluated polymanuronates on the differentiation of 3T3-L1 adipocytes. Polymannuronates increased glucose utilization and reduced the accumulation of triglycerides in the cells. The differentiation showed the same results as Oil red O staining. Also, the polymannuronates inhibited GPDH activity as a result of the restrained adipogenesis promotion process in 3T3-L1 adipocytes. The addition of the differentiation promotion factor to 3T3-L1 promoted the differentiation of adipocytes and increased the expression of the leptin level. However the addition of polymannuronates inhibited differentiation of adipocytes and the leptin secretion level in cells by checking the leptin protein level in the culture media. As well as this, it also inhibited the transcriptional mechanism and leptin mRNA expression. These results suggest that the addition of polymannuronates improves the physiological function of 3T3-L1 cells by reducing the accumulation of triglyceride and GPDH activity, and the repressing expression of leptin at a molecular level.

• Ocean and Polar Research
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• v.44 no.1
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• pp.61-67
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• 2022

In this study, in order to evaluate the anti-obesity effect of sargassum horneri extract, the effects of the extract on lipase activity and preadipocyte differentiation in 3T3-L1 cells were investigated. S. horneri extract between 0.0 and 1.0 mg/mL showed no cytotoxicity and inhibited lipase activity by 68.1%. When S. horneri extract was utilized at levels of 0.25, 0.5, and 1.0 mg/mL in 3T3-L1 cells, preadipocytes differentiation decreased by 11.4, 19.7, and 25.6%, respectively, showing anti-obesity effects. In addition, after treatment with 1.0 mg/mL S. horneri extract, the mRNA expression levels of sterol regulatory element binding proteins-1c (SREBP-1c), peroxisome proliferator activated receptor-γ (PPAR-γ), CCAAT enhancer binding protein-α (CEBP-α), fatty acid synthase (FAS), and stearoyl-CoA desaturase1 (SCD1) in 3T3-L1 cells were significantly decreased (p < 0.05) by 65.2, 54.9, 50.0, 33.8, and 33.8% respectively. These results showed that S. horneri extract suppresses lipase activity and prophylactic preadipocyte differentiation in 3T3-L1, and thus can be used as an anti-obesity agent in functional foods and medicines.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2613-2618
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• 2014

Aims: Dysfunction of the host immune system in cancer patients can be due to a number of factors, including lymphocyte apoptosis. Several studies showed that $Foxp3^+T$ cells take part in inducing this process by expressing FasL in tumor patients. However, the relationship between apoptosis, $CD8^+T$ cells and $Foxp3^+T$ cells in HCC patients is still unclear. The present study was designed to investigate the correlation between apoptosis levels and Fas/FasL expression in $CD8^+T$ lymphocytes and $Foxp3^+T$ cells in patients with HCC. Methods: $CD8^+T$ cells and $CD3^+Foxp3^+T$ cells were tested from peripheral blood of HCC patients and normal controls and subjected to multicolor flow cytometry. The expression of an apoptosis marker (annexin V) and the death receptor Fas in $CD8^+T$ cells and FasL in $CD3^+Foxp3^+T$ cells were evaluated. Serum TGF-${\beta}1$ levels in patients with HCC were measured by enzyme-linked immunosorbent assay. The relationship between apoptosis and Fas expression, as well as FasL expression in $CD3^+Foxp3^+T$ cells was then evaluated. Results: The frequency of $CD8^+T$ cells binding annexin V and Fas expression in $CD8^+T$ cells, were all higher in HCC patients than normal controls and the proportion of apoptotic $CD8^+T$ cells correlated with their Fas expression. Serum TGF-${\beta}1$ levels correlated inversely with $CD3^+Foxp3^+T$ cells. Conclusions: Fas/FasL interactions might lead to excessive turnover of $CD8^+T$ cells and reduce anti-tumor immune responses in patients with HCC. Further investigations of apoptosis induction in $Fas^+CD8^+T$ cells in vitro are required.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1062-1068
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• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.