• Journal of Life Science
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• v.31 no.1
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• pp.1-9
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• 2021

Brown seaweeds have been shown to decrease blood glucose levels and improve insulin sensitivity previously. In this study, we investigated the effect of fucoidan, a complex polysaccharide derived from brown seaweeds, on glucose uptake to improve insulin resistance, and examined its mechanism of action in 3T3-L1 adipocytes. We observed that fucoidan significantly increased glucose uptake and it was related to an increased expression of plasma membrane-glucose transporter 4 (PM-GLUT4) in 3T3-L1 adipocytes. Fucoidan treatment increased the activation of phosphatidylinositol-3-kinase (PI3K) and the phosphorylation of insulin receptor substrate 1 (IRS1tyr) compared with that of the control cells. Fucoidan also promoted the phosphorylation of Akt and protein kinase C (PKC)-λ/ζ compared to that of the control cells. Moreover, fucoidan significantly upregulated acetyl-CoA-carboxylase (ACC) and adenosine monophosphate - activated protein kinase (AMPK) phosphorylation. As a result, translocation of GLUT4 was significantly enhanced in 3T3-L1 adipocytes, which significantly promoted glucose uptake via the PI3K/AMPK pathways. The elevation of glucose uptake by fucoidan was blocked by inhibitor of PI3K and inhibitor of AMPK in 3T3-L1 adipocytes. These findings indicate that fucoidan might ameliorate glucose uptake through GLUT4 translocation to the plasma membrane by activating the PI3K/Akt and AMPK pathways in 3T3-L1 adipocytes. Fucoidan is thought to be of high material value to diabetes treatments and functional foods.

• 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.

• Proceedings of the Korea Water Resources Association Conference
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• 2016.05a
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• pp.524-524
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• 2016

본 연구에서는 홍천 자운지구 고랭지 농업지역을 대상으로 장기간의 유역조사와 하천 모니터링을 통해 관측된 축척 데이터를 이용하여 비점오염저감 효과를 정량화하고 추후 모니터링 자료로서 탁수와 비점오염원 저감시설의 저감효과와 비점오염원의 효율적인 관리를 위한 기초자료를 제공하고자 하였다. 모니터링 결과 소양호 유역의 오염부하(1차 강우사상) 중 홍천군 자운지구의 유역단위 비점오염 저감효과의 분석에는 강우량과 단위면적당 오염부하를 이용하였으며, 자운천은 SS 5,396,761 kg, COD 82,261 kg, BOD 57,329 kg, T-N 68,711 kg, T-P 3,091 kg이었으며, 오염부하(2차 강우사상)는 SS 320,293 kg, COD 34,588 kg, BOD 22,350 kg, T-N 48,954 kg, T-P 640 kg으로 나타났다. 또한 소양호 유역의 EMC(1차 강우사상) 중 자운천은 SS 829.9 mg/L, COD 12.7 mg/L, BOD 8.8 mg/L, T-N 10.567 mg/L, T-P 0.475 mg/L 이었으며, EMC(2차 강우사상)는 SS 68.6 mg/L, COD 7.4 mg/L, BOD 4.8 mg/L, T-N 10.487 mg/L, T-P 0.137 mg/L로 나타났다. 소양호 유역의 단위면적당 오염부하(1차 강우사상) 중 자운천은 SS 402.0 kg/ha/event, COD 6.1 kg/ha/event, BOD 4.3 kg/ha/event, T-N 5.118 kg/ha/event, T-P 0.230 kg/ha/event 이었으며, 오염부하(2차 강우사상)는 SS 23.9 kg/ha/event, COD 2.6 kg/ha/event, BOD 1.7 kg/ha/event, T-N 3.646 kg/ha/event, T-P 0.048 kg/ha/event로 나타났다. 오염부하에서는 1차 강우에 비해 2차 강우에서는 SS 5,076,468 mg/L, COD 47,673 mg/L, BOD 34,979 mg/L, T-N 19,757 mg/L, T-P 2451 mg/L로 1차 강우사상에 비해 오염부하가 낮은 것으로 나타났다. 또한 EMC에서는 1차 강우에 비해 2차 강우에서는 SS 761.3 mg/L, COD 5.3 mg/L, BOD 4.0 mg/L, T-N 0.080 mg/L, T-P 0.338 mg/L로 1차 강우사상에 비해 EMC가 저감되었다. 단위면적당 오염부하는 1차 강우에 비해 2차 강우에서는 SS 378.1 mg/L, COD 3.5 mg/L, BOD 2.6 mg/L, T-N 1.472 mg/L, T-P 0.182 mg/L로 1차 강우사상에 비해 EMC가 저감되었다. 강우량과 강우강도 그리고 영농활동의 시기에 따른 EMC와 단위면적당 오염부하는 큰 차이를 보였으며, 예년에 비해 적은 강수량으로 인해 탁수와 비점오염부하의 배출이 상대적으로 적었던 것으로 판단된다. 소양호 유역과 같은 넓은 유역에서 시행되는 비점오염원의 저감연구는 오랜 시간 동안 자료를 축적해야 유의미한 평가가 이루어질 수 있으므로 장기적이고 지속적인 모니터링을 통한 유량 데이터 자료구축과 수질분석뿐만 아니라, 발생되는 비점오염 물질의 관리를 위한 체계적인 연구가 지속되어야하며, 아울러 농민의 인식도 변화 등을 포함하는 많은 인자들을 정밀히 조사하고 다각도로 분석하여 저감효과에 대한 지속적인 연구가 필요하다.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.582-590
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• 2015

In the present study, we investigated the effect of 70% EtOH extract from Hippophae Rhamnoides L. leaves (HRL) on the anti-obesity effect in 3T3-L1 cells. The effects of HRL on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. In addition, we examined the gene expression levels by using RT-PCR and western blot. The results of this analysis showed that 100 ㎍/㎖ HRL significantly increased the inhibition of lipid accumulation by 82.25%; significantly decreased the mRNA expression of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα), and fatty acid synthase (FAS) in 3T3-L1 cells as well as the stimulated protein expression of AMP-activated protein kinase (AMPK); and suppressed the expression level of PPARγ. These results suggest that HRL can prevent adipogenesis through activation of AMPKα and inhibition of adipogenesis transcription factors.

• Preventive Nutrition and Food Science
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• v.9 no.4
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• pp.336-341
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• 2004

Leptin, resisitn and PAI-1 (plasminogen activator inhibitor-1) are synthesized and secreted by rodent fat cells and recently postulated to be an important link to obesity. This study was conducted to characterize the hormonal regulation of leptin, resistin, and PAI-1 gene expression in the 3T3-L1 adipocytes. The cells were treated with 0.5 $\mu$M insulin, 1 $\mu$M dexamethasone (Dex), or 0.05 $\mu$M triiodothyronine (T3) for 72 hours. The mRNA levels of each peptide were measured by semi-quantitative RT-PCR. The mRNA level of the leptin-producing ob gene was significantly increased by insulin, Dex, and T3 by 3.2-, 3.1- and 2.7-fold, respectively, compared to the control (p < 0.05). The level of resistin mRNA was increased by insulin, Dex, and T3 by 2.7-, 2.5- and 2-fold, respectively, compared to the control (p < 0.05). Likewise, the level of PAI-1 mRNA was significantly increased by insulin, Dex, and T3 compared to the control (p < 0.05). Taken together, our results suggest that insulin, Dex, and T3 may regulate the gene expression of leptin, resistin, and PAI-1 in 3T3-L1 adipocytes.

• Journal of Life Science
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• v.32 no.10
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• pp.762-770
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• 2022

Hyperglycemia in type 2 diabetes can be alleviated by promoting cellular glucose uptake. Betulinic acid (3β,-3-hydroxy-lup-20(29)-en-28-oic acid) is a pentacyclic lupane-type triterpenoid compound. Although there have been studies on the antidiabetic activity of betulinic acid, studies on cellular glucose uptake are lacking. We investigated the effects of betulinic acid on glucose uptake and its mechanism of action in 3T3-L1 adipocytes. Betulinic acid significantly stimulated glucose uptake in 3T3-L1 adipocytes by increasing the phosphorylation of the insulin receptor substrate 1-tyrosine (IRS-1tyr) in the insulin signaling pathway, which in turn stimulated the activation of phosphoinositide 3-kinase (PI3K) and the phosphorylation of protein kinase B (Akt). The activation of PI3K and Akt by betulinic acid translocated glucose transporter 4 to the plasma membrane (PM-GLUT4), thereby increasing the expression of PM-GLUT4 and thus stimulating cellular glucose uptake. Betulinic acid also significantly increased the phosphorylation/activation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase. The activation of PI3K and AMPK by betulinic acid was confirmed using the PI3K inhibitor wortmannin and the AMPK inhibitor compound C. The increase in glucose uptake induced by betulinic acid was significantly decreased by wortmannin and compound C in the 3T3-L1 adipocytes. These results suggest that betulinic acid stimulates glucose uptake by activating PI3K and AMPK in 3T3-L1 adipocytes.

• Journal of the Korean Society of Food Culture
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• v.34 no.1
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• pp.84-92
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• 2019

Purpose: The objective of this study was conducted to investigate the effects of rutin, buckwheat components on cell growth and anti-inflammation in adipocyte 3T3-L1 and human colon cancer cell SW-480. Methods: We cultured 3T3-L1 adipocyte and SW-480 colon cancer cell to confluence, at which time starvation was induced with SFM for 1 day. Cells were then cultured in medium containing 0, 25, 50, or $100{\mu}mol/mL$ of rutin 3T3-L1 or 0, 10, 20, or $40{\mu}mol/mL$ SW-480. Cell viability was measured using a cell viability kit. In addition, we examined the expression of mRNA related to inflammation. RT-PCR was used to quantity tumor necrosis factor ($TNF-{\alpha}$), interleukin-$1{\beta}$ ($IL-1{\beta}$), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) mRNA levels. Results: Rutin significantly inhibited 3T3-L1 and SW-480 cell proliferation in a dose and time dependent manner. Rutin also significantly reduced the mRNA expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ at the highest dose. In addition, rutin treatment caused a significant reduction in COX-2 and iNOS mRNA levels compared to the control group. Conclusion: Overall, our results suggest that rutin has the potential to reduce inflammation, and that these effects are greater during tissue-damaging inflammatory conditions.

• Journal of Life Science
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• v.33 no.11
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• pp.859-867
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• 2023

Lupeol is a type of pentacyclic triterpene that has been reported to have therapeutic effects for treating many diseases; however, its effect on insulin resistance is unclear clear. This study examined the inhibitory effect of lupeol on the serine phosphorylation of insulin receptor substrate-1 in insulin resistance-induced 3T3-L1 adipocytes. 3T3-L1 cells were cultured and treated with tumor necrosis factor-α (TNF-α) for 24 hours to induce insulin resistance. Cells treated with different concentrations of lupeol (15 μM or 30 μM) or 100 nM of rosiglitazone were incubated. Then, lysed cells underwent western blotting. Lupeol exhibited a positive effect on the negative regulator of insulin signaling and inflammation-activated protein kinase caused by TNF-α in adipocytes. Lupeol inhibited the activation of protein tyrosine phosphatase-1B (PTP-1B)-a negative regulator of insulin signaling-and c-Jun N-terminal kinase (JNK); it was also an inhibitor of nuclear factor kappa-B kinase (IKK) and inflammation-activated protein kinases. In addition, Lupeol downregulated serine phosphorylation and upregulated tyrosine phosphorylation in insulin receptor substrate-1. Then, the downregulated phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway was activated, the translocation of glucose transporter type 4 was stimulated to the cell membrane, and intracellular glucose uptake increased in the insulin resistance-induced 3T3-L1 adipocytes. Lupeol may improve TNF-α-induced insulin resistance by downregulating the serine phosphorylation of insulin receptor substrate 1 by inhibiting negative regulators of insulin signaling and inflammation-activated protein kinases in 3T3-L1 adipocytes.

• Journal of Life Science
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• v.32 no.7
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• pp.542-549
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• 2022

Polymethoxyflavones (PMFs) are flavonoids mainly found in citrus fruits and have been reported to exhibit a wide range of bioactivities, including anti-obesity, anti-cancer, and anti-inflammatory actions. To utilize PMFs as functional materials, it is necessary to develop a simple method of obtaining PMFs from citrus tissues containing a large amount of PMFs. It has been reported that Jinkyool (C. sunki Hort ex. Tanaka) peel contained a large amount of PMFs, but there are no studies on PMFs isolated from its leaves. In this study, we established a simple procedure for obtaining the PMF-rich fraction (PRF) from the leaves of Jinkyool and investigated the effects of PRF on lipid metabolism in 3T3-L1 cells. PRF inhibited lipogenesis during the differentiation of 3T3-L1 preadipocytes. It decreased the expression of peroxisome proliferator-activated receptor gamma (PPAR𝛾) and CCAAT/enhancer binding protein alpha (CEBP𝛼), FAS, and adipocyte fatty-acid-binding protein 2 (aP2). In mature 3T3-L1 adipocytes, PRF increases the phosphorylation of protein kinase A (PKA)/hormone-sensitive lipase (HSL), which are key factors involved in lipolysis. Moreover, it increases the phosphorylation of the AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) involved in fatty acid oxidation. These results suggest that PRF from Jinkyool leaves can be used as an anti-obesity agent with the action of inhibiting lipogenesis and promoting lipolysis and fatty acid oxidation in 3T3-L1 adipocytes.

• Korean Journal of Pharmacognosy
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• v.42 no.4
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• pp.302-308
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• 2011

As a part of our ongoing program on finding biologically active components from natural source we found three known constituents from the EtOH extract of the wheat bran. The known compounds were identified as tachioside (1), pinellic acid (2) and tryptophan (3). The structure and relative stereochemistry were determined from MS, 1D and extensive 2D NMR techniques as well as by comparison of their data with the published values. All isolates were tested their inhibitory effects on the adipogenesis in 3T3-L1 cells. The effect of compounds from wheat bran on 3T3-L1 adipocyte differentiation were measured by Oil Red O staining. These results demonstrate that tachioside (1) and pinellic acid (2) decreased lipid content in 3T3-L1 adipocytes by inhibiting lipogenesis. These compounds had shown antiobesity activities.