• Title/Summary/Keyword: oil red-O staining

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Protective effect of Buddha's Temple extract against tert-butyl hydroperoxide stimulation-induced oxidative stress in DF-1 cells

  • Eun Hye Park;Sung-Jo Kim
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1120-1129
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    • 2023
  • Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.

Effect of silver (Ag) deposition before physical developer to develop latent fingerprints on wet paper (젖은 종이류에서 잠재지문을 현출하기 위한 physical developer 전 은 (Ag) 증착의 효과)

  • Wonyoung Lee;Min-Jeong Kang;Yoo-Jung Lee;Je-seol Yu
    • Analytical Science and Technology
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    • v.36 no.6
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    • pp.324-331
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    • 2023
  • Latent fingerprints on wet paper cannot be developed using amino acid reaction reagents. Therefore, physical developer (PD) or lipid staining reagents like oil red O (ORO) should be utilized. However, ORO is not very effective in developing fingerprints that are older than approximately 4 weeks. On the other hand, PD performs well in developing older fingerprints, but it cannot do so for relatively fresh fingerprints. Additionally, PD has the disadvantage of taking a long time to develop fingerprints. In this study, in order to overcome the limitations of PD, we attempted to increase its reactivity by applying Ag-PD, which involves depositing silver onto paper using vacuum metal deposition (VMD), and compared this with fingerprints developed using ORO and PD. As a result, Ag-PD showed superior fingerprint development compared to ORO and PD on wet paper stored for 2~8 weeks, and the fingerprint development time for PD was significantly reduced to 90~150 seconds.

Inhibition of Lipid Accumulation in 3T3-L1 Adipocytes by Different Enzymatic Hydrolysates of Dried Red Sea Cucumber Stichopus japonicus (건조 방법에 따른 홍해삼(Stipchopus japonicus) 효소 가수분해물의 지방 축적 억제 효과)

  • Kim, Seo-Young;Oh, Jae-Young;Kim, Eun-A;Heo, Soo-Jin;Kim, Kil-Nam;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.5
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    • pp.707-716
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    • 2020
  • Red sea cucumber Stichopus japonicus, was dried using three methods-far-infrared ray, vacuum, and freeze drying and then enzymatically hydrolyzed using nine proteases: Alcalase, Flavourzyme, Kojizyme, Neutrase, Protamex, trypsin, α-chymotrypsin, and papain. In addition, the potential ability of hydrolysates to inhibit lipid accumulation in 3T3-L1 adipocytes was evaluated. The yield of hydrolysates from red sea cucumbers dried using each method was higher than that of the distilled water extract, and protein contents were either similar or higher. The hydrolysates that exhibited inhibitory effects on lipid accumulation, as demonstrated via Oil red O staining, were those obtained by far-infrared ray drying coupled with Alcalase, Flavourzyme, Kojizyme, or Neutrase treatment. In addition to the advantages of far-infrared drying and the characteristics of Flavourzyme, the Flavourzyme hydrolysate of far-infrared-dried red sea cucumber showed the highest inhibitory effect on lipid accumulation. In addition, this hydrolysate significantly decreased the expression of the protein factor fatty acid-binding protein 4, which is related to the late differentiation of 3T3-L1 adipocytes. Taken together, these results suggest that Flavourzyme hydrolysates from farinfrared-dried red sea cucumber may be used as a functional food and/or a pharmaceutical ingredient for the inhibition of lipid accumulation.

Anti-adipogenic Activity of Acer tegmentosum and its Constituent, Catechin in 3T3-L1 Cells

  • Liu, Qing;Shin, Eun-Jin;Ahn, Mi-Jeong;Hwang, Bang-Yeon;Lee, Mi-Kyeong
    • Natural Product Sciences
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    • v.17 no.3
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    • pp.212-215
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    • 2011
  • In the course of screening anti-adipogenic activity of natural products employing the preadipocyte cell line, 3T3-L1 as an in vitro assay system, the EtOAc fraction of the stem barks of Acer tegmentosum Maxim (Aceraceae) showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Activity-guided fractionation led to the isolation of active constituent, (+)-catechin. (+)-Catechin showed inhibitory activity on adipocyte differentiation in dose-dependent manner. Further studies with interval treatment demonstrated that (+)-catechin exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Our present study also showed that (+)-catechin significantly inhibited the preadipocyte proliferation. Taken together, these results suggest that (+)-catechin, a constituent of A. tegmentosum might contribute the anti-adipogenic activity of A. tegmentosum.

Effect of Foilum mori on Adipocyte Differentiation

  • Lee, Geun-Won;Byun, Sang-Yo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.5
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    • pp.299-302
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    • 2003
  • Several natural products were tested to control the differentiation of cultured human mesenchymal stem cell into adipocyte. The extent of the inhibitory effect on the conversion of adipose was measured using Oil red O staining, which stains accumulated lipid droplets in the cytoplasm of adipocyte. Of the various natural product extracts, the adipocyte differentiation was inhibited by an extract from Folium mori in the concentration range 1${\times}$10$\^$-4/∼5${\times}$l0$\^$-2/ g/mL. These results suggest that Folium mori has an inhibitory activity toward the adipose conversion, which is a major cause of obesity.

Synergic Effect on Adipocytic Differentiation by a Combination of Thyroxine andGlucocorticoid in A549 Cells (A549 폐암세포주의 지방세포 분화에 미치는 티록신 및 당질 코르티코이드 호르몬의 상승 효과)

  • Hyeon-Ji Kim;Sun-Ha Moon;Sung-Ho Lee;Byeong-Gyun Jeon
    • Journal of Life Science
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    • v.33 no.9
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    • pp.693-702
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    • 2023
  • The present study examined the rate of cell growth and differentiation potential into adipocytes in A549 lung adenocarcinoma cells exposed to each adipogenic medium containing glucose metabolism hormones, such as thyroxine (T4) thyroid hormone and glucocorticoid (GC) adrenal steroid hormone, as well as pioglitazone (PGZ), a PPARγ agonist. Following each adipogenic treatment for 2 weeks, the rate of cell growth was significantly (p<0.05) inhibited, and the level of telomerase activity was significantly (p<0.05) decreased in the PGZ-based adipogenic medium containing both T4 and GC hormone compared with those containing each T4 or GC hormone. Moreover, the adiposome-like vesicles were highly reacted with Oil-Red O staining solution, and the levels of transcripts expressed in the differentiating adipocytes for adipogenesis, including adinopectin, leptin, and resistin, were significantly (p<0.05) increased in the PGZ-based adipogenic medium containing both T4 and GC hormone compared with those of the adipogenic medium containing each T4 or GC hormone, implying that adipocytic differentiation has fully occurred in the A549 cancer cells. Based on present observations, the PGZ-based adipogenic medium containing both T4 and GC efficiently induces inhibition of cell growth and cellular differentiation into adipocytes in A549 cancer cells rather than in the adipogenic medium containing only T4 or GC hormone. Adipogenic treatment could provide potential probability in cancer chemotherapy.

Anti-obesity effect of resveratrol-amplified grape skin extracts on 3T3-L1 adipocytes differentiation

  • Zhang, Xian-Hua;Huang, Bo;Choi, Soo-Kyong;Seo, Jung-Sook
    • Nutrition Research and Practice
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    • v.6 no.4
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    • pp.286-293
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    • 2012
  • Resveratrol (3,4,5-trihydroxy-trans-stilbene), a phytoalexin found in grape skin, grape products, and peanuts as well as red wine, has been reported to have various biological and pharmacological properties. The purpose of this study was to investigate the anti-obesity effect of resveratrol-amplified grape skin extracts on adipocytes. The anti-obesity effects of grape skin extracts were investigated by measuring proliferation and differentiation in 3T3-L1 cells. The effect of grape skin ethanol extracts on cell proliferation was detected by the MTS assay. The morphological changes and degree of adipogenesis of preadipocyte 3T3-L1 cells were measured by Oil Red-O staining assay. Treatment with extracts of resveratrol-amplified grape skin decreased lipid accumulation and glycerol-3-phosphate dehydrogenase activity without affecting 3T3-L1 cell viability. Grape skin extract treatment resulted in significantly attenuated expression of key adipogenic transcription factors, including peroxisome proliferator-activated receptor, CCAAT/enhancer-binding proteins, and their target genes (FAS, aP2, SCD-1, and LPL). These results indicate that resveratrol-amplified grape skin extracts may be useful for preventing obesity by regulating lipid metabolism.

Comparison of Dipogenic and Osteogenic Potentials of Visceral and Subcutaneous Adipose Stem Cells of Human (인체 피하지방과 내장지방에서 유래한 줄기세포의 지방분화 및 골분화 능력 차이)

  • Kang, Min Gu;Park, Jong Lim;Lee, Jin Hee;Chang, Hak;Minn, Kyung Won;Park, Gyu Ju
    • Archives of Plastic Surgery
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    • v.34 no.5
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    • pp.537-542
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    • 2007
  • Purpose: Adipose tissue contains a population of pluripotent stem cells capable of differentiating along multiple mesenchymal cell lineages. It is well known that fat depots from different part of our body shows different nature not only in morphological aspect but also physiologic aspect. The authors compared the adipogenic potentials and osteogenic potentials of adipose stem cells from different anatomical sites of human. Methods: After laparotomy by surgery team, the authors isolated these adipose stem cells successfully from 7 men with an average age of 58, and induced differentiation along adipogenic and osteogenic lineages in vitro. On the 14th day, cells cultured in adipogenic media differentiated into adipocytes in vitro, as evidenced by positive Oil Red O staining of lipid vacuoles. On the 21st day, cells cultured in osteogenic media differentiated into osteoblasts in vitro as demonstrated by Alizarin red staining of a calcified extracellular matrix. Results: After exposure to adipogenic and osteogenic differentiation medium, subcutaneous adipose stem cells were found to possess greater adipogenic and osteogenic potentials than cells isolated from visceral adipose tissues. Conclusion: This study indicates that adipogenic and osteogenic potentials of adipose stem cells vary by their anatomical sites, with subcutaneous adipose stem cells exhibiting higher adipogenic and osteogenic potential than those isolated from visceral fat.

Tanshinone I, an Active Ingredient of Salvia miltiorrhiza, Inhibits Differentiation of 3T3-L1 Preadipocytes and Lipid Accumulation in Zebrafish

  • Kwon, Hyo-Shin;Jang, Byeong-Churl
    • Journal of Korean Medicine for Obesity Research
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    • v.20 no.2
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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.

Protective Effects of Korean Red Ginseng against Alcohol-induced Hepatosteatosis (알코올에 의해 유발된 지방변성증에서 홍삼의 보호효과)

  • Kim, Sun Ju;Ki, Sung Hwan;Lee, Sangkyu
    • Journal of Life Science
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    • v.25 no.3
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    • pp.317-322
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    • 2015
  • Alcohol-induced fatty liver (steatosis) results from excessive generation of reducing equivalents by ethanol metabolism. Generally, chronic ethanol treatment causes hepatosteatosis by regulating sterol regulatory element-binding protein 1c (SREBP-1c), which increases the synthesis of hepatic lipids. The effect of ethanol on SREBP-1c is mediated through mammalian sirtuin-1 (SIRT-1), a NAD+-dependent protein deacetylase that regulates hepatic lipid metabolism. Ginseng is a widely used herbal medicine that is used in Asia for its anti-diabetes and anti-obesity effects. The pharmacological and therapeutic effects of ginseng are primarily produced by bioactive constituents known as ginsenosides. Here, we examined the regulatory effects of Korean red ginseng (KRG) extracts on SREBP-1c and SIRT-1 on lipid homeostasis in AML-12 mouse hepatocytes. AML-12 cells were treated with ethanol and/or KRG extracts (0 - 1,000 μg/ml). Lipid droplets were assayed using Oil red O staining, and western blotting was used to measure SIRT-1 and SREBP-1 expression. Treatment with KRG extracts restored SIRT-1 expression and reduced SREBP-1c expression in ethanol-treated cells. We also showed that KRG extract and ginsenosides Rb2 and Rd significantly decreased SREBP-1 acetylation in ethanol-treated cells. These results show that treatment with KRG extract and its active ginsenoside constituents Rb2 and Rd protected against alcohol-related hepatosteatosis via regulation of SIRT-1 and downstream acetylation of SREBP-1c, which altered hepatic lipid metabolism.