• 대한족부족관절학회지
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• 제18권3호
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• pp.100-107
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• 2014

Purpose: The purpose of this study is to evaluate the efficacy of mesenchymal stem cell (MSC) isolation by the magnetic-activated cell sorting (MACS) method in tendon tissue-derived cells compared to the colony picking method for isolation of MSCs by picking colony-forming cells. Materials and Methods: Human tendon-derived cells were isolated by enzyme digestion using normal tendon tissues from three donors. We used the magnetic kit and well-known MSC markers (CD90 or CD105) to isolate MSCs in tendon-derived cells using MACS. Cloning cylinders were used to isolate colony-forming cells having MSC characteristics in tendon-derived cells. Colony-forming unit-fibroblast (CFU-F) assay was used to evaluate the self-renewal capacity of cells isolated using the colony picking method or MACS. For comparison of differentiation potentials into osteogenic or adipogenic lineage between two groups, alizarin red S and oil red O staining were performed at 14 days after induction of differentiation in vitro. Results: Flow cytometry results showed that early passage tendon-derived cells expressed CD44 in 99.13%, CD90 in 56.51%, and CD105 in 86.19%. In the CFU-F assay, CD90+ or CD105+ cells isolated with MACS showed larger colony formation in size than cells isolated using the colony picking method. We also observed that CD90+ or CD105+ cells were constantly differentiated into both osteogenic and adipogenic lineages in cells from all donors, whereas cells isolated using the colony picking method were heterogeneous in differentiation potentials to the osteogenic and adipogenic lineages. Conclusion: CD90+ or CD105+ cells isolated using MACS showed superior MSC characteristics in the self-renewal and multi-differentiation capacities compared with cells isolated using the colony picking method.

• 대한한방내과학회지
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• 제39권6호
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• pp.1181-1190
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• 2018

Objective: This study investigated the effect of purified medical herb extracts such as Alisma canaliculatum and Polyporus umbellatuson adipogenic differentiation of human bone marrow derived mesenchymal stromal stem cells (hBMSCs). Methods: Two different medical herb were extracted using hot distilled water. The optimal concentration of extracts were fixed at 100 ng/ml by means of cell viability and cytotoxic assay. To test the adipogenic differentiation ability of extracts, we induced the adipogenesis of hBMSCs for 21 days. At day 5, the cell was harvested to check mRNA and protein expression level of adipogenic related factors. The efficacy of lipid droplet formation was evaluated using the oil-red O staining method at days 21. Results: Two different medical herb extracts have no toxicity on hBMSCs. And two different medical herb extracts significantly decreased formation of lipid droplet compared with control groups in hBMSCs. The A. canaliculatum extract group showed the lowest mRNA and protein expression level of adipossgenic related transcription factors. This data suggests that extract of A. canaliculatum and P. umbellata decrease the adipogenic differentiation of hBMSCs. Conclusions: Our findings indicate that water-extract of A. canaliculatum and P. umbellata will be useful therapeutic reagents for prevention of obesity related disease such as diabetes, hyperlipidemia, coronary artery disease, and osteoporosis.

• Nutrition Research and Practice
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• 제15권2호
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• pp.160-172
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• 2021

BACKGROUND/OBJECTIVES: Nutritional status and food intake during pregnancy and lactation can affect fetal programming. In the current metabolic syndrome epidemic, high-fructose diets have been strongly implicated. This study investigated the effect of maternal high-fructose intake during pregnancy and lactation on the development of metabolic syndrome in adult offspring. SUBJECTS/METHODS: Drinking water with or without 20% fructose was administered to female C57BL/6J mice over the course of their pregnancy and lactation periods. After weaning, pups ate regular chow. Accu-Chek Performa was used to measure glucose levels, and a tail-cuff method was used to examine systolic blood pressure. Animals were sacrificed at 7 months, their livers were excised, and sections were stained with Oil Red O and hematoxylin and eosin (H&E) staining. Kidneys were collected for gene expression analysis using quantitative real-time Polymerase chain reaction. RESULTS: Adult offspring exposed to maternal high-fructose intake during pregnancy and lactation presented with heavier body weights, fattier livers, and broader areas under the curve in glucose tolerance test values than control offspring. Serum levels of alanine aminotransferase, aspartate aminotransferase, glucose, triglycerides, and total cholesterol and systolic blood pressure in the maternal high-fructose group were higher than that in controls. However, there were no significant differences in mRNA expressions of renin-angiotensin-aldosterone system genes and sodium transporter genes. CONCLUSIONS: These results suggest that maternal high-fructose intake during pregnancy and lactation induces metabolic syndrome with hyperglycemia, hypertension, and dyslipidemia in adult offspring.

• 한방비만학회지
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• 제21권2호
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• pp.69-79
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• 2021

Objectives: This study aimed to compare the anti-obesity effects of raw and pickled garlic in vitro in 3T3-L1 preadipocytes. Methods: The pickled garlic samples comprised the following: garlic aged in vinegar (VG), garlic aged in soy sauce, and vinegar (1:1, v/v) (PG) and raw garlic (RG) as control. Hexane, butanol, and distilled water were used to prepare the fractions. The pancreatic lipase inhibitory activity was used as a measure of anti-obesity effects of the extracts. The lipid droplet accumulation and triglyceride content in the 3T3-L1 cells were measured using Oil red O staining and triglyceride assay kits, respectively. The adipogenesis related protein expression levels were analyzed using the kits and the western blot method. Results: The pancreatic lipase inhibitory activity of the garlic extracts (VG, PG, RG) was the highest in the butanol fraction, and the inhibitory effect was the highest in RG, followed by PG and VG. All garlic butanol extracts suppressed triglyceride accumulation in differentiated adipocytes (P<0.05) through the activation of cyclic adenosine monophosphate (AMP), AMP-activated protein kinase, carnitine palmitoyl transferase-1, and the inhibition of fatty acid synthase. Raw garlic extracts significantly inhibited the expression of proteins involved in adipogenesis as compared to pickled garlic. Conclusions: Raw garlic has the potential to be an effective natural material for reducing obesity compared to pickled garlic with vinegar or soy sauce.

• Journal of Nutrition and Health
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• 제17권4호
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• pp.253-261
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• 1984

식이중 지방 수준과 fiber 종류가 흰 쥐의 혈청과 간 조직내의 지질 함량에 미치는 영향을 알아보기 위해 지방 수준을 0% , 5% , 20% 수준으로 달리 하였으며 fiber 종류는 고지방 식이군( 20% 지방 )에 pectin과 cellulose를 각각 10% 수준으로 첨가하였다. 또 식이내 cholesterol첨가의 효과를 관찰하기 위해 고지방 식이군에 0.5% cholesterol을 첨가한 후 pectin과 cellulose를 10%씩 넣어 전체적으로 7군으로 나누어 각 각 다른 식이로 4주간 사양한 후 혈청과 간에서 지질 함량을 조사하였으며 간 조직의 형태학적 조사를 실시하여 다음과 같은 결과를 얻었다. 1) 식이 섭취량은 지방 수준이 낮을수록 높았으나 뚜렷한 차이는 보이지 않았으며 pectin과 cellulose 첨가로 인해서 섭취량이 줄었다. 증체량은 지방 수준이나 fiber종류에 따라 큰 차이를 보이지 않았다. 변의 무게는 지방 수준에 따른 차이는 없었으나 fiber 첨가로 인해서는 pectin은 약간의 감소를 cellulose 는 뚜렷한 증가를 보였다. 또 cholesterol을 첨가한 군 간의 비교에서도 pectin첨가군은 감소를 cellulose첨가군은 증가를 나타냈다. 간과 epididymal fat pad의 무게는 지방 수준이 높아짐에 따라 증가하였으며 fiber 종류에 따라서는 pectin첨가군이 cellulose 첨가 군 보다 낮았다. 2) 혈청 분석 결과는 식이내 지방 수준이 높을수록 total lipids수준이 높아지는 경향을 보였으며 pectin이 cellulose보다 혈청내 지질 수준을 뚜렷하게 낮추었다. Total cholesterol 및 phospholipids 량에 있어서도 지방 수준이 증가함에 따라 높아졌으며 cellulose보다 pectin이 첨가 되었을때 현저한 감소를 나타냈다. HDL- cholesterol수준은 무지방군에서는 매우 높게 나타났으나 표준군과 고지방군간에 는 명확한 결과를 얻을 수 없었다. 3) 간 분석 결과는 지방수준이 높을수록 total lipids량이 높아지는 경향을 보였다. 또 pectin 첨가가 cellulose첨가군보다 현저히 낮은 lipids수준을 나타냈다. 간의 total cholesterol함량은 지방 수준이 높아짐에 따라 증가하였으나 뚜렷한 차이는 아니었으며 pectin첨가가 cellulose첨가보다 cholesterol 수준이 상당히 낮았다. 4) 간 조직의 현미경 관찰 결과는 지방 수준이 높아짐에 따라 조직내 지방 침착이 심해졌으며 fiber 의 첨가로 인해서는 pectin은 뚜렷한 지방 침착의 감소를 보였으나 cellulose는 거의 감소 효과를 보이지 않았다.

• 동의생리병리학회지
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• 제30권2호
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• pp.88-94
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• 2016

The study was carried out to examine the anti-obesity effects of 40% ethanol extract from green tea seed (GS) and quantitative determination of caffeine as its major compound. The specificity was satisfied with retention time and UV spectrum by analysis of caffeine using HPLC and comparison with standard compound. It showed a high linearity in the calibration curve with a coefficient of correlation (R²) of 0.9974. The amount of caffeine in GS was about 4.649 mg/g (0.465%) in the three times analysis, and relative standard deviation (RSD) was less than 0.452% by the validated method. The anti-obesity effects of GS were evaluated by using Oil Red O staining in 3T3-L1 adipocytes and body weight, visceral fat and lipid profiles in high fat diet (HFD)-induced C57BL/6 obese mice. Our results indicated that treatment with GS dose-dependently decreased lipid accumulation contents (p<0.001). Moreover, after oral administration for 30 days feeding with HFD-induced obses mice, GS (100 and 300 mg/kg/day) produced a significant decrease in serum total cholesterol (TC), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and visceral fat. Thus, the result of this study indicate that the GS may be a useful resource for the management of obesity.

• Animal Bioscience
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• 제35권5호
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• pp.763-777
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• 2022

Objective: Excessive lipid accumulation in adipocytes results in prevalence of obesity and metabolic syndrome. Curcumin (CUR), a naturally phenolic active ingredient, has been shown to have lipid-lowering effects. However, its underlying mechanisms have remained largely unknown. Therefore, the study aims to determine the effect of CUR on cellular lipid accumulation in porcine subcutaneous preadipocytes (PSPA) and to clarify novel mechanisms. Methods: The PSPA were cultured and treated with or without CUR. Both cell counting Kit-8 and lactate dehydrogenase release assays were used to examine cytotoxicity. Intracellular lipid contents were measured by oil-red-o staining extraction and triglyceride quantification. Apoptosis was determined by flow cytometry and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay. Adipogenic and apoptosis genes were analyzed by quantitative polymerase chain reaction and Western blot. Results: The CUR dose-dependently reduced the proliferation and lipid accumulation of PSPA. Noncytotoxic doses of CUR (10 to 20 μM) significantly inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and expression of adipogenic genes peroxisome proliferation-activity receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α, sterol regulatory element-binding protein-1c, adipocyte protein-2, glucose transporter-4 as well as key lipogenic enzymes fatty acid synthase and acetyl-CoA carboxylase, while ERK1/2 activation significantly reversed CUR-reduced lipid accumulation by increasing PPAR-γ. Furthermore, compared with differentiation induced media treated cells, higher dose of CUR (30 μM) significantly decreased the expression of AKT and B-cell lymphoma-2 (BCL-2), while increased the expression of BCL-2-associated X (BAX) and the BAX/BCL-2 expression ratio, suggesting triggered apoptosis by inactivating AKT and increasing BAX/BCL-2 ratio and Caspase-3 expression. Moreover, AKT activation significantly rescued CUR inhibiting lipid accumulation via repressing apoptosis. Conclusion: These results demonstrate that CUR is capable of suppressing differentiation by inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis via decreasing AKT and subsequently increasing BAX/BCL-2 ratio and Caspase-3, suggesting that CUR provides an important method for the reduction of porcine body fat, as well as the prevention and treatment of human obesity.

• 대한본초학회지
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• 제29권2호
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• pp.15-21
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• 2014

Objectives : The purpose of this study was to investigate inhibitory effects of steamed Polygonatum odoratum extract (POE) on differentiation and adipogenesis in 3T3-L1 adipocytes. Methods : Polygonatum odoratum (P. odoratum) extract was extracted with ethyl acetate. Total phenolic and flavonoid contents in POE were measured for antioxidant activity. The spectrophotometric method was used to determine the DPPH and ABTS radical scavenging activity and ferric-reducing antioxidant potential (FRAP). MTT assay was examined for cell toxicity, oil red O staining was performed for intracelluar adipogenesis in differentiated 3T3-L1 adipocytes. Western blot analysis for measurement of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor${\gamma}$ ($PPAR{\gamma}$) and AMP-activated protein kinase (AMPK) expressions were performed. Results : The results revealed that POE has antioxidant activities. Contents of total polyphenolics and flavonoids were $50.83{\pm}1.52$ GAE mg/100g dry weight of POE and $17.05{\pm}2.47$ RE mg/100g dry weight of POE, respectively. DPPH radical scavenging activity, and FRAP in 10 mg/ml concentration were $92.1{\pm}0.6%$, $244.8{\pm}9.0{\mu}M$ Fe(II) and ABTS inhibition in 5 mg/ml concentration was $84.8{\pm}4.1%$. Treatment of POE in adipocytes inhibited the differentiation and adipogenesis of 3T3-L1 adipocytes compared to those of vehicle control. Additionally, protein expressions of $C/EBP{\alpha}$ and $PPAR{\gamma}$, major transcription factor for the adipogenic genes, were significantly decreased compared to those of vehicle control (p<0.05). Futhermore, phosphorylation of AMPK was increased in 3T3-L1 adipocytes treated with POE compared to that of vehicle control (p<0.05). Conclusions : we demonstrate that steamed P. odoratum extract (POE) has potentiating antioxidant activities, inhibits differentiation and lipid accumulation and also induces energy expenditure in adipocytes, which may contribute to antiobesity property.