• 한국식생활문화학회지
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• 제33권5호
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• pp.464-471
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• 2018

Purpose: This study examined the effects of ${\alpha}$-lipoic acid in diluted solvents on cell growth in 3T3-L1 cells according to the treated concentration and times. Methods: Adipocyte 3T3-L1 cell were cultured. Confluent cells underwent starvation with SFM for 1 day and then were cultured in a medium containing various concentrations 0, 100, 200, and $400{\mu}mol/L$ of ${\alpha}$-lipoic acid. The cell viability was measured using the EZ Cytox assay kit. In addition, the effect of ${\alpha}$-lipoic acid of diluted solvents on the cell growth in 3T3-L1cells was examined according to the treated concentration and times. Results: The ${\alpha}$-lipoic acid diluted ethanol inhibited cell proliferation in a dose and time dependent manner. The ${\alpha}$-lipoic acid diluted ethanol induced adipocyte 3T3-L1 cells proliferation with an adipocyte inducer. In addition, ${\alpha}$-lipoic acid inhibited adipocyte 3T3-L1 growth in a dose and time dependent manner (p<0.05). Conclusion: This study showed that a treatment with ${\alpha}$-lipoic acid diluted ethanol inhibits cell growth of, adipocyte 3T3-L1 cells induced with an adipocyte inducer, ($200{\mu}mol/L$ of ${\alpha}$-lipoic acid) treated for 48 hr.

• 한국식생활문화학회지
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• 제34권1호
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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.

• 한국수산과학회지
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• 제33권6호
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• pp.541-545
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• 2000

알긴산은 3T3-L1 세포의 분화를 억제하였다. 3T3-L1 세포의 분화를 촉진시키는 인자로 밝혀진 IGF-I과 insulin을 이용하여 알긴산의 분화에 미치는 영향을 검토한 결과, 알긴산은 insulin의 분화촉진작용을 특이적으로 억제하였다. 본 연구는 알긴산을 이용하여 알긴산의 지질 감소효과를 세포의 수준에서 검증하고자 하였다는 점에서 그 의의가 있으며, 이에 알긴산의 분화억제효과가 일어나는 메카니즘에 관한 연구가 있어야 할 것으로 생각된다.

• 동아시아식생활학회지
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• 제25권6호
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• pp.990-998
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• 2015

The aim of this study was to examine the cytotoxicity and potential inhibitory effects from four types of edible domestic brown seaweeds, Undaria pinnatifida (UP), Laminaria japonica (LJ), Sargassum fulvellum (SF), and Hizikia fusiforme (HF), on preadipocyte differentiation in 3T3-L1 cell line. Water and ethanol extracts from the four types of seaweeds were prepared and tested for cell viability in the 3T3-L1 cell line by using MTT assay. In addition, various doses of the water extract of seaweeds (WES) and ethanol extract of seaweeds (EES) were treated at the beginning of 3T3-L1 differentiation and continued until the cells were fully differentiated to adipocytes. Oil Red-O staining was performed to determine the potential cell differentiation inhibitory effects of the WES and EES by measuring the levels of lipid droplet accumulation in 3T3-L1 adipocytes. $PPAR{\gamma}$ mRNA expression levels were significantly reduced by WESs of UP, LJ, and HF as well as EESs of LJ and HF. As a result, we observed the superior cell differentiation inhibitory effects of WES compared to that of EES in a dose-dependent manner without any significant cytotoxicity in mouse adipocytes.

• Journal of Animal Science and Technology
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• 제55권1호
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• pp.19-23
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• 2013

본 연구는 콜라비가 돼지 지방전구세포와 $_3T_3-L_1$ 세포의 증식과 분화에 미치는 영향을 구명하기 위해 수행하였다. 돼지 지방전구세포는 신생자돈의 등지방에서 분리했다. 세포를 접종한 1일 후에 세척했고(day 0), 세포증식에 미치는 영향을 구명하기 위해서 2일 동안(day 0~day 2) 25 ng/ml과 100 ng/ml의 콜라비 알코올 추출물(과피와 과육)를 처리했다. 세포분화를 구명하기 위해서는 DMEM/F- 12 배지에 6일 동안(day 0~day 6) 배양하고 배양초기 2일 동안(day 0~day 2) 콜라비를 처리하고 day 6에 세포 분화를 측정했다. 콜라비 과피 25 ng/ml와 100 ng/ml은 돼지 지방전구세포의 증식을 각각 4.59%, 17.7% 억제했고, 콜라비 과육은 각각 11.4%, 19.2% 억제했다. 반면 돼지 지방전구세포의 분화는 억제하지 않았다. 콜라비가 $_3T_3-L_1$ cell의 증식과 분화에 미치는 작용을 구명하기 위해, 돼지 지방전구세포처럼, 세포 배양초기 2일간 콜라비를 처리했는데 콜라비 과피와 과육 둘 다 세포의 증식과 분화에 영향을 미치지 않았다. 본 연구의 결과를 요약하면, 콜라비는 돼지 지방전구세포의 증식을 억제했으나 분화는 억제하지 않았고, 한편 $_3T_3-L_1$ cell의 증식과 분화 모두 영향을 미치지 않았다.

• 대한의용생체공학회:의공학회지
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• 제39권4호
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• pp.168-174
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• 2018

Adipocytes affect obesity through the regulation of lipid metabolism. Physical loading is an important regulator of fat tissue. There are ongoing in vitro studies inducing mechanotransduction on 3T3-L1 preadipocytes with mechanical stimulus in order to treat obesity by inhibiting adipogenesis and provoking cell death. In this study, our goal was to suggest a new therapy for obesity by investigating whether fluid shear stress (FSS) changes transcription factors on 3T3-L1 related with adipogenesis and cell death. FSS loading was applied to 3T3-L1 preadipocytes at 1Pa and 1Hz. After loading, bright field images were taken and an immunofluorescence assay was conducted to observe actin stress fiber formation. Western blot analysis was conducted to identify the activation of the ERK pathway as well as the adipogenic factors, which including C/EBPs and $PPAR{\gamma}$. The expression of osteopontin, a protein related to inflammation in adipose tissue, and cell death related factors, Bax, Bcl-2, and Beclin, were also measured. Results showed that FSS stimulated the formation of actin stress fibers in 3T3-L1 and also that the activation of C/EBPs decreased significantly when compared with the control group. $PPAR{\gamma}$ activation in the 2 hour FSS group was lower than the 1 hour FSS group, which implied that the results were time dependent. Additionally, there were no differences in the expression of cell death factors after FSS loading. In summary, similar to other fibroblasts, the formation of actin stress fibers induced by mechanotransduction may affect the differentiation of 3T3-L1, leading to inhibition of adipogenesis and inflammation.

• 약학회지
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• 제39권5호
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• pp.535-540
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• 1995

The purpose of this research was to investigate effects of glycyrrhizin on the differentiation of preadipocytes, 3T3-Ll cells and to characterize the action of glycyrrhizin that affect the responses of 3T3-Ll cells during differentiation. The differentiation of 3T3-Ll cells was stimulated by glycyrrhizin, and triglyceride contents was increased in the differentiated 3T3-LI cell extracts. Total protein contents was increased by glycyrrhizin or inductive agents in the differentiated 3T3-Ll cell extracts. Calmodulin contents was increased by inductive agents, but the contents was not affected by glycyrrhizin in the differentiated 3T3-Ll cell extracts. The results suggest that glycyrrhizin has a stimulating activity of adipose conversion, but the activity is not related to calmodulin contents during the process of differentiation of 3T3-LI cells.

• 한국식품영양과학회지
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• 제39권1호
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• pp.31-35
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• 2010

본 실험에서는 hamster $\beta$-cell인 HIT-T15 cell을 이용하여 돼지감자추출물의 생리활성 및 기능을 검증하고자 하였다. 돼지감자추출물을 첨가한 NC(0 ${\muL/mL$), HT2(1.1 ${\muL/mL$), HT3(1.5 ${\muL/mL$)군과 inulin을 첨가한 NC(0 ${\muL/mL$), IN2(1.8 ${\muL/mL$), IN3(2.5 ${\muL/mL$)군으로 나누어 실험하였다. 세포 viability 측정한 결과 시료를 첨가하지 않은 군을 100%로 보았을 때 돼지감자추출물을 첨가한 HT3(1.5 ${\muL/mL$)군과 inulin을 첨가한 IN2(1.8 ${\muL/mL$), IN3(2.5 ${\muL/mL$) 군에서 세포생존율이 유의적으로 증가하였다(p<0.05). 시료처리 후 췌장 $\beta$-세포 파괴를 유도하지 않고 HIT-T15 cell의 cell culture supernatant를 이용하여 cytotoxicity를 측정한 결과 시료를 첨가하지 않은 NC(0${\muL/mL$)군에 비해 모든 군에서 cytotoxicity가 낮게 나타났다. Alloxan(4 mM)으로 $\beta$-세포 파괴를 유도하여 HIT-T15 cell에서 세포보호 효과를 측정한 결과 시료를 첨가하지 않은 NC(0 ${\muL/mL$)군에 비해 돼지감자추출물을 첨가한 HT2(1.1 ${\muL/mL$), HT3(1.5 ${\muL/mL$)군에서 세포생존율이 유의적으로 증가하였다(p<0.05). 또한 췌장 $\beta$-세포 파괴를 유도하여 HIT-T15 cell이 분비한 인슐린 분비능 및 세포 내 $NAD^+$/NADH 함량을 측정한 결과 시료를 첨가하지 않은 NC(0 ${\muL/mL$)군에 비해 돼지감자추출물을 첨가한 HT3(1.5 ${\muL/mL$)군에서 인슐린 분비량과 $NAD^+$/NADH 함량이 유의적으로 증가하였다(p<0.05). 이상의 연구 결과 돼지감자추출물은 HIT-T15 cell의 생존율을 높이고, 세포보호 효과를 가짐으로써 인슐린 분비능 정상화 및 $NAD^+$ 함량을 증가시켜 혈당 조절 및 당뇨에 긍정적 효과가 있을 것으로 사료된다.

• Journal of Animal Science and Technology
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• 제54권2호
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• pp.103-109
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• 2012

본 연구는 기존 단독배양 위주로 이루어져온 세포배양 연구의 방법학적 한계의 극복과 대안을 제시하고자 지방과 근육세포주의 단독 및 공동배양에서 배양기법에 따른 지방 및 근육세포의 분화에 미치는 영향을 비교 조사하고자 실시하였다. 3T3-L1 (지방세포) 및 L6 (근육세포) 세포주는 성장배지인 10% FBS/DMEM (1% Pen-Strep solution 및 0.1% Fungizone 첨가) 하에서 48h 동안 단독배양 후 5% FBS/DMEM에서 배양하였다. 분화를 위한 단독 및 공동배양에서는 지방 및 근육세포 모두 분화유도물질 없이 2% FBS/DMEM으로 배양하였고, 공동배양에서는 $0.4{\mu}m$ insert membrane을 사용하여 6 well plate 하단에 L6 cell을, 상단에는 3T3-L1 cell을 공생시켰다. 지방 및 근육세포 분화정도 측정은 세포별 형태학적 측정과 glycerol-3-phosphate dehydrogenase (GPDH) 및 creatine kinase (CK) 분석을 통해 조사되었다. 형태학적으로 볼때 3T3-L1 세포주는 공동배양보다 단독배양 시 분화가 더욱 잘 일어났고 L6 세포주의 경우 역으로 같았다. 세포물질 분석에서는 분화배지 처리일(day 0)과 비교해 단독 및 공동배양 모두 지방세포 내 GPDH의 활성도가 유의적으로(P<0.05) 증가했음을 확인할 수 있었고 단독배양이 공동배양보다 유의적으로(P<0.05) 높은 수준의 GPDH 활성도를 보였다. L6 역시 마찬가지로 분화배지 처리일에 비하여 단독 및 공동배양 모두 CK 활성도가 유의적으로(P<0.05) 높았고, CK 활성도가 공동배양에서 유의적으로(P<0.05) 높게 나타남을 확인할 수 있었다. 이러한 결과는 기존 연구에서 이용된 단독 배양을 통한 세포 분화 결과 등은 생체와 비교 시 방법학적 한계로 인해 실제 생체 내에서는 그 분화정도가 매우 다를 것으로 생각되며, 이것은 앞으로 정확한 세포배양 결과 확보를 위해서는 단독배양보다는 공동배양기법을 사용해야 함을 의미한다. 향후 다양한 조건과 분화조절 물질들의 첨가를 통한 추가적인 공동배양실험이나 지방분화관련 분자생물학적 물질분석 등 다양한 실험 수행 시 보다 현실적이고 대량의 기초자료 확보가 가능할 것으로 판단된다.

• 대한본초학회지
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• 제31권4호
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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.