• 한방비만학회지
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• 제20권2호
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• pp.109-121
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• 2020

Objectives: Tanshinone I is a bioactive constituent in Salvia miltiorrhiza. At present, the anti-obesity effect and mechanism of tanshinone I are not fully understood. Here we investigated the effect of tanshinone I on lipid accumulation in 3T3-L1 preadipocytes and zebrafish. Methods: Lipid accumulation and triglyceride (TG) content in 3T3-L1 cells were determined by Oil Red O staining and AdipoRed assay, respectively. The expression and phosphorylation levels of adipogenic/lipogenic proteins in 3T3-L1 cells were evaluated by Western blotting. The messenger RNA (mRNA) expression levels of adipogenic/lipogenic markers and leptin in 3T3-L1 cells were measured by reverse transcription polymerase chain reaction (RT-PCR). Lipid accumulation in zebrafish was assessed by LipidGreen2 staining. Results: Tanshinone I at 5 μM largely blocked lipid accumulation and reduced TG content in differentiating 3T3-L1 cells. Furthermore, tanshinone I decreased the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and perilipin A but also the phosphorylation of signal transducer and activator of transcription-3 (STAT-3) in differentiating 3T3-L1 cells. In addition, tanshinone I increased the phosphorylation of adenosine 3',5'-cyclic monophosphate (cAMP)-activated protein kinase (AMPK) while decreased the intracellular adenosine triphosphate (ATP) content with no change in the phosphorylation and expression of liver kinase-B1 in differentiating 3T3-L1 cells. Importantly, tanshinone I also reduced the extent of lipid deposit formation in developing zebrafish. Conclusions: These findings demonstrate that tanshinone I has strong anti-adipogenic effects on 3T3-L1 cells and reduces adiposity in zebrafish, and these anti-adipogenic effect in 3T3-L1 cells are mediated through control of C/EBP-α, PPAR-γ, STAT-3, FAS, ACC, perilipin A, and AMPK.

• 생명과학회지
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• 제25권3호
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• pp.317-322
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• 2015

알코올에 의한 지방간(지방변성증)은 에탄올 대사에 의해 감소되는 당량의 과도한 발생에 의해 유발된다. 일반적으로 만성적인 에탄올 투여는 간 지질의 합성을 증가시키는 sterol regulatory element-binding protein 1c (SREBP-1c)를 조절함으로써 지방변성증을 유발시킨다. SPEBP-1c에서 에탄올의 영향은 간에서의 지방대사를 조절하는 NAD⁺ 의존적 단백질 탈아세틸화효소인 mammalian sirtuin-1 (SIRT-1)에 의해 조정된다. 홍삼은 항당뇨와 항비만 효과를 위해 아시아에서 광범위하게 사용되는 한약재이다. 홍삼의 약리학적 치료학적인 효과는 진세노사이드와 같은 생물활성 성분에 의해 주로 일어난다. 따라서 우리는 마우스 간세포주인 AML-12 세포에서 SREBP-1과 SIRT-1에대한 한국홍삼 추출물의 조절효과를 평가하였다. 알코올과 홍삼추출물(0-1,000 μg/ml)을 AML-12 세포주에 처리하고, 지방소립을 Oil red O 염색법으로 확인하고, western blots을 사용해 SIRT-1과 SREBP-1의 발현을 확인하였다. 에탄올을 처리한 세포에서 홍삼추출물은 SIRT-1과 SREBP-1c의 발현을 회복시켰다. 또한 에탄올이 처리된 세포에서 홍삼추출물과 진세노사이드 Rb₂와 Rd가 SREBP-1을 유의적으로 감소시키는 것으로 확인 되었다. 결과적으로 홍삼과 활성 진세노사이드 성분인 Rb₂와 Rd가 SIRT-1과 간 지질대사를 변화시키는 SREBP-1c의 아세틸화의 조절을 통해 알코올에 의한 간지방변성을 억제하는 것을 확인하였다.

• 한방재활의학과학회지
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• 제24권3호
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• pp.11-27
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• 2014

Objectives This study was designed to evaluate the effects of Ephedra Sinica and Fibrosum Gypsum extract on obesity by using 3T3-L1 cells and high fat diet rats. Methods In vitro, Ephedra Sinica and Fibrosum Gypsum extract (50, 100, 200, $500{\mu}g/ml$) were added in 3T3-L1 cells. Cytotoxicity was measured by MTT assasy. Adipocyte differentiation was measured by Oil Red O staining, GPDH activity and $C/EBP{\alpha}$ protein expression. In vivo, Sprague-Dawley rats were divided into 5 groups : Normal diet group (Normal group), taken high fat diet and no treatment group (Control group), taken high fat diet and orally administered Ephedra Sinica and Fibrosum Gypsum extract daily (Group I: 50 mg/kg, Group II: 100 mg/kg, Group III : 200 mg/kg, oral). For 6 weeks of administration, body weight and the amount of food intake were measured once a week. After administration, blood analysis (AST, ALT, T-Bilirubin, BUN, RBC, Hb, HCT), serum lipid level (triglyceride, Total cholesterol, HDL, LDL), serum leptin level, epididymal adipose tissue weight and histological finding of liver were estimated. Results In vitro, The cytotoxicity was not significant. 3T3-L1 cell's differentiation was significantly decreased in Oil Red O staining, GPDH activity and $C/EBP{\alpha}$ protein expression. In vivo, Body weight and the amount of food intake, AST, ALT, Total cholesterol, TG, LDL, serum leptin, epididymal adipose tissue weight showed significant decrease in group I, group II and group III. There were no significant difference in T-bilirubin, BUN, RBC, Hb and HCT between all groups. HDL showed significant increase in group I, group II and group III. In histological finding of liver tissue, there were decreased adiposity and cytopathic effect in group I, group II and group III. Conclusions It is suggested that Ephedra Sinica and Fibrosum Gypsum extract can be used in the treatment of obesity.

• Preventive Nutrition and Food Science
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• 제11권1호
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• pp.31-35
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• 2006

In order to improve the antiobesity effect of Kochujang, 1% of sea tangle powder, alginic acid extract, and fucoidan extract were added to Kochujang. Sea tangle powder-added Kochujang decreased leptin secretion by only 12% compared to Kochujang, whereas alginic acid or fucoidan-added Kochujang significantly decreased leptin secretion by more than 60% in 3T3-L1 adipocytes. Fucoidan, one of the active components of sea tangle, decreased leptin secretion by 56%, 60%, and 60% compared to the control in the concentrations of $1{\mu}M,\;2.5{\mu}M,\;and\;5{\mu}M$, respectively. To see the effect of fucoidan on TG formation during adipocyte differentiation, 3T3-L1 cells were treated with $1{\mu}M\;and\;5{\mu}M$ concentrations of fucoidan during adipocyte differentiation (from 'day 0' to 'day 6'). Oil red O staining showed fucoidan decreased the amount of TG droplets and $5{\mu}M$ fucoidan potently inhibited TG formation. To see the effect of fucoidan on lipolysis, differentiated 3T3-L1 adipocytes were treated with fucoidan. The secretion of glycerol, which is used to measure lipolytic activity, was increased by 21%, 37%, and 53% compared to the control in the concentrations of $1{\mu}M,\;2.5{\mu}M,\;and\;5{\mu}M$, respectively. Oil red O staining showed fucoidan decreased TG amount at $1{\mu}M\;and\;5{\mu}M$ concentrations. These results suggest that fucoidan decreases leptin secretion and TG accumulation by inhibition of adipocyte differentiation and induction of lipolysis. Since fucoidan is reported to have various biological activities in addition to an anti-adipogenic effect, it seems valuable to develop fucoidan-added Kochujang as a multi-functional Kochujang.

• 생명과학회지
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• 제22권8호
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• pp.1052-1056
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• 2012

미역(Undaria pinnatifada)은 낮은 칼로리 및 요오드의 원료로써 천연체중조절식품으로 알려져 있다. 미역이 체중조절식품으로 알려져 있음에도 불구하고, 지방세포 분화 및 지방축적에 관한 저해 기작은 연구가 미비하다. 본 연구에서는 3T3-L1에서 지방세포로 분화가 일어나는 단계에서 미역에탄올추출물의 효과 및 기작을 확인하였다. 미역에탄올추출물의 독성과 지방축적저해효과는 MTT assay, Oil red O staining, RT-PCR과 western blot으로 분석하였다. 미역에탄올추출물은 50 ${\mu}g/ml$의 농도에서 독성을 띄지 않았다. 3T3-L1의 분화 및 지방세포에서 triglyceride축적과정동안 50 ${\mu}g/ml$의 미역에탄올추출물을 처리하였으며, 미역에탄올추출물은 지방세포에서 triglyceride의 축적을 40% 감소시켰다. 지방세포 특이적 단백질인 Peroxisome proliferator activated receptor ${\gamma}$ ($PPAR{\gamma}$), leptin과 Hormone sensitive lipase (HSL)의 발현은 RT-PCR과 western blot으로 확인하였다. $PPAR{\gamma}$의 과발현은 지방세포의 분화를 촉진시킨다. 또한 지방세포 크기의 증가와 세포 내 triglyceride의 함량에 따라 leptin은 세포 외로 분비된다. 그러므로 $PPAR{\gamma}$와 leptin은 비만의 지표로 사용된다. 첨가한 미역에탄올추출물의 농도가 높아질수록 $PPAR{\gamma}$와 leptin의 발현이 억제되었다. 이상의 결과를 통하여, 미역의 에탄올 추출물은 지방전구세포의 분화를 억제시키며, 지방세포 내 triglyceride의 축적을 저해하는 것으로 판단된다.

• 생명과학회지
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• 제29권2호
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• pp.209-214
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• 2019

본 연구에서는 괭생이모자반 에탄올 추출물의 항비만 효과에 대해 알아보고자 3T3-L1 preadipocyte에서 Oil Red O staining과 triglyceride content, adipogenesis의 발현에 대해 조사하였다. MTT assay를 통해 괭생이모자반 에탄올 추출물의 세포 독성을 확인 해 본 결과 $200{\sim}1,000{\mu}g/ml$ 농도에서 90% 이상의 세포 생존율을 확인 할 수 있었다. 또한 insulin, dexamethasone과 IBMX 복합물을 처리하여 분화를 유도하였을 때는 분화유도제를 처리하지 않은 군에 비해 활발하게 지방구를 형성하였으나, 괭생이모자반 에탄올 추출물을 250, 500, $1,000{\mu}g/ml$ 농도로 처리하였을 때는 농도 의존적으로 지방구 형성을 억제하였다. 이를 정량하여 triglyceride content (%)로 나타낸 결과 농도별로 13%, 16%, 23% 억제 효과를 나타내었다. 또한 Western blot assay를 통해 adipogenic transcription factor의 발현양상을 비교해 본 결과, 분화유도제 처리에 의해 $C/EBP{\alpha}$, $C/EBP{\beta}$와 $PPAR{\gamma}$의 발현이 증가하였으나, 괭생이모자반 에탄올 추출물을 처리하였을 때는 이러한 전사인자들의 발현이 농도 의존적으로 감소됨을 확인 할 수 있었다. 이상의 결과를 통해 괭생이모자반 에탄올 추출물은 adipogenic transcription factor의 발현을 조절하여 지방세포의 분화를 억제하였으며, 지방구와 triglyceride 생성을 감소시켜 최종적으로 비만억제 효과를 나타낸다는 것을 알 수 있었다.

• Journal of Nutrition and Health
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• 제52권1호
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• pp.17-25
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• 2019

본 연구에서는 댕댕이나무 열매 추출물이 3T3-L1과 마우스 지방유래줄기세포의 지방 분화유도 및 지방생성에 미치는 영향을 살펴보았다. 3T3-L1에 댕댕이나무 열매 추출물을 처리하였을 때, 농도의존적으로 지방구의 생성을 줄였고 지방세포 분화에 있어서 중요한 전사인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$, SREBP1의 발현을 억제시켜 지방 합성이 감소됨을 확인하였다. 또한, 마우스 지방에서 분리한 줄기세포의 지방 분화과정에서도 댕댕이나무 열매 추출물이 $PPAR{\gamma}$, $C/EBP{\alpha}$, SREBP1의 단백질 발현을 감소시켜 지방 축적을 농도 의존적으로 억제하였다. 이상의 결과로 댕댕이나무 열매 추출물은 세포독성이 없는 농도에서 지방 세포의 분화를 억제 하는 것으로 확인되어 항비만 기능성 소재로서의 활용 가능성이 있을 것으로 사료된다.

• 대한본초학회지
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• 제33권2호
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• pp.53-58
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• 2018

Objectives : Plantaginis asiaticae Folium (PA) has been widely used in Korean medicine for treatment of liver disease, stomach troubles and inflammation. We investigated the effect of PA on lipid accumulation in 3T3-L1 according to extraction conditions. Methods : The MTT assay was employed to evaluate the cytotoxicity of PA extracted by different solvents (water, 50% ethanol, and 95% ethanol) on 3T3-L1 preadipocytes. Oil red O staining was used to identify intracellular lipid accumulation in 3T3-L1. 3T3-L1 adipocytes were treated with PA at concentration ranging in 0.1, 0.2 and $0.4mg/m{\ell}$. PA was extracted by different extraction conditions such as extraction solvents, extraction time, and extraction temperature. In addition, UPLC analysis was used for determination of candidates of active ingredients in PA. Results : 3T3-L1 preadipocytes were treated with PA extracted by different solvents (water, 50% ethanol, and 95% ethanol) and there was no cytotoxicity. Oil red O staining was employed to identify the effect of PA on lipid accumulation in 3T3-L1. In the present study, PA water extraction at $70^{\circ}C$ for 6 hours decreased greatly in lipid accumulation. The range of concentrations was 0.1, 0.2 and $0.4mg/m{\ell}$. Concentration at $0.2mg/m{\ell}$ was the most effective one among them. Candidates of active ingredients in PA were shown plantamajoside and acteoside through UPLC. Conclusions : These results suggest that the effect of PA water extraction at $70^{\circ}C$ on lipid accumulation in 3T3-L1 is superior to other extraction conditions. We suppose that plantamajoside and acteoside may be candidates of active ingredients in PA.

• 대한본초학회지
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• 제39권4호
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• pp.29-35
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• 2024

Objectives : Obesity is an imbalance between energy intake and consumption due to overeating and lack of exercise, and if it persists, it increases non-infectious diseases such as high blood pressure, diabetes, and hyperlipidemia. In this study, we tried to investigate the possibility of using Chamaeneron angustifolia (CA) as a material for anti-obesity by confirming the effect of inhibiting lipid differentiation. Methods : We measured the effects of CA extract on oil-red-o staining, cell cytotoxicity evaluation activity using 3T3-L1 cells. Additionally, we assessed fat decomposition and metabolism-related protein expression through Western blot analysis. Results : In this study, the anti-obesity effects of CA extract were experimentally assessed. Results showed significant inhibition of adipocyte differentiation and accumulation at concentrations of 0.05, 0.1, and 0.2 mg/ml of oil-red-o staining, with reductions of 80% or more. CA notably increased the phosphorylation of AMPK protein expression compared to the control group across all concentrations. Additionally, phosphorylation of ACC significantly increased at a concentration of 0.2 mg/ml compared to the control. PPAR-γ, which regulates adipogenesis, exhibited a significant decrease compared to the control, while protein expression of CPT-1, involved in fatty acid oxidation, showed a concentration-dependent increase across all groups. Therefore, CA extract demonstrates potential as a functional material for anti-obesity by increasing the expression of proteins related to fat decomposition and synthesis while decreasing others. Conclusions : These results suggest that CA may also be useful as an anti-obesity functional substance.

• Journal of Ginseng Research
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• 제42권4호
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• pp.419-428
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• 2018

Background: Despite the large number of studies on ginseng, pharmacological activities of ginseng seed oil (GSO) have not been established. GSO is rich in unsaturated fatty acids, mostly oleic and linoleic acids. Unsaturated fatty acids are known to exert a therapeutic effect in nonalcoholic fatty liver disease (NAFLD). In this study, we investigated the protective effect and underlying mechanisms of GSO against NAFLD using in vitro and in vivo models. Methods: In vitro lipid accumulation was induced by free fatty acid mixture in HepG2 cells and by 3 wk of high fat diet (HFD)-feeding in Sprague-Dawley rats prior to hepatocyte isolation. The effects of GSO against diet-induced hepatic steatosis were further examined in C57BL/6J mice fed a HFD for 12 wk. Results: Oil Red O staining and intracellular triglyceride levels showed marked accumulation of lipid droplets in both HepG2 cells and rat hepatocytes, and these were attenuated by GSO treatment. In HFD-fed mice, GSO improved HFD-induced dyslipidemia and hepatic insulin resistance. Increased hepatic lipid contents were observed in HFD-fed mice and it was lowered in GSO (500 mg/kg)-treated mice by 26.4% which was evident in histological analysis. Pathway analysis of hepatic global gene expression indicated that GSO increased the expression of genes associated with ${\beta}$-oxidation (Ppara, Ppargc1a, Sirt1, and Cpt1a) and decreased the expression of lipogenic genes (Srebf1 and Mlxipl), and these were confirmed with reverse transcription and quantitative polymerase-chain reaction. Conclusion: These findings suggest that GSO has a beneficial effect on NAFLD through the suppression of lipogenesis and stimulation of fatty acid degradation pathway.