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.
The pathological features of a mass in the back skin region of an 8-year-old castrated male dog are described herein. The cut section of the tumor was white to tan with a soft multilobulated mass containing hemorrhagic and necrotic foci and a mucinous-like composition. Microscopically, the tumor was composed of a mixture of lipocytes, lipoblasts, spindle cells and stellate cells and had a myxoid background. Oil red O staining revealed that the cytoplasm of neoplastic cells contained large numbers of lipid droplets. Immunohistochemically, tumor cells were positive for vimentin and S-100 protein. The skin mass was diagnosed as myxoid liposarcoma.
Journal of the Korean Society of Food Science and Nutrition
/
v.44
no.9
/
pp.1264-1269
/
2015
To evaluate the functional effect of ornithine produced by isolated lactic acid bacteria, we examined the anti-lipid accumulation effect of ornithine produced by isolate lactic acid bacteria on 3T3-L1 cells. Lactic acid bacteria (Pediococcus strain) were isolated from nuruk, which is made from wheat, rice, and barley (whole grain, grits, or flour) by fermenting microorganisms (Aspergillus, Rhizopus, and yeasts). Pediococcus strain was identified by 16S rDNA sequencing analyses, and cells were collected by centrifugation and developed as an ornithine starter. makgeolli, an ornithine-containing Korean traditional alcoholic beverage, was made with isolated lactic acid bacteria and arginine. makgeolli was made with the help of ornithine starter using a makgeolli making kit. We evaluated the anti-proliferation effect of ornithine makgeolli on 3T3-L1 cells. To determine the anti-proliferation effect of ornithine makgeolli on preadipocytes, lipid droplets were quantified and stained with Oil Red O. makgeolli made with ornithine starter and arginine showed a 3-fold higher concentration of ornithine compared to makgeolli without starter and arginine. In the results of 3T3-L1 cell line experiment, lipid accumulation was significantly reduced by adding 0.05 mg/mL of ornithine makgeolli compare to the control (adipocyte without sample). In conclusion, ornithine makgeolli containing ornithine starter isolated from nuruk showed an anti-lipid accumulation effect with increased ornithine content without toxicity.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.7
/
pp.895-900
/
2012
The effects of three fractions, hexane (BHHH), chloroform (BHHC), and ethyl acetate (BHHE), from water extract of Benincasa hispida on the underlying mechanisms of adipogenesis were investigated in 3T3-L1 cells. Intracellular lipid droplets were stained with Oil Red O dye and quantified. Compared to control, lipid accumulation significantly decreased by 11% and 13% upon treatment with BHHC and BHHE, respectively at a concentration of 50 ${\mu}g/mL$. Intracellular triglyceride (TG) levels were also reduced by 21% and 16%, respectively, at the same concentration. To determine the mechanism behind the reductions in TG content and lipid accumulation, glycerol release and expression levels of adipogenic marker genes were measured. The levels of free glycerol released into culture medium increased by 13% and 17% upon treatment with BHHC and BHHE, respectively. In subsequent measurements using real-time polymerization chain reaction, the mRNA levels of $PPAR{\gamma}$, C/$EBP{\alpha}$, and leptin significantly decreased upon treatment with BHHE (45%, 67%, and 35%) in comparison with non-treated control. These results suggest that BHHE inhibits adipocyte differentiation by blocking $PPAR{\gamma}$, C/$EBP{\alpha}$, and leptin gene expression in 3T3-L1 cells, resulting in reduced lipid accumulation, increased glycerol release, and intracellular triglycerides.
Ultrastructural changes of the pericarp in Citrus reticulata has been investigated during hesperidium abscission. The pericarp was composed of compactly arranged parenchyma cell layers during early stages of fruit development. The outermost exocarp was green and active in photosynthesis. However, cells in the exocarp soon changed into collenchyma cells by developing unevenly thickened walls within a short time frame. As the fruit approached maturation, the chlorophyll gradually disappeared and chloroplasts were transformed into carotenoid-rich chromoplasts. In the mature fruit the exocarp consisted of large, lobed collenchyma cells with primary pit fields and numerous plasmodesmata. The immature mesocarp was a relatively hard and thick layer, located directly under the exocarp. With development, the deeper layers of the exocarp merged into the white, spongy mesocarp. Before separation of the hesperidium from the plant, some unusual features were detected in the plasma membrane of the exocarp cells. The number of small vacuoles and dark, irregular osmiophilic lipid bodies also increased enormously in the exocarp collenchyma after the abscission. They occurred between the plasma membrane and the wall, and invaginated pockets of the plasma membrane containing double-membraned vesicles were also frequently noticed. The lipid bodies in the cytoplasm were often associated with other organelles, especially with plastids and mitochondria. The plastids, which were irregular or amoeboid in shape, contained numerous large lipid droplets, and occasional clusters of phytoferritin, as well as few loosely -oriented peripheral lamellae. Myelin-like configurations of membrane were frequently observed in the vacuoles, as was the association of lipid bodies with the vacuolar membrane. Most vacuoles had an irregular outline, and lipid bodies were often connected to the tonoplast of the vacuoles. The structural changes underlying developmental, particularly to senescence, processes in various hesperidium will be reported in the separate paper.
Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.
Min, Byulchorong;Lee, Heejin;Song, Ji Hye;Han, Myung Joo;Chung, Jayong
Nutrition Research and Practice
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v.8
no.6
/
pp.655-661
/
2014
BACKGROUND/OBJECTIVES: The purpose of this study was to examine the effects and associated mechanisms of arctiin, a lignan compound found in burdock, on adipogenesis in 3T3-L1 cells. Also, the effects of arctiin supplementation in obese mice fed a high-fat diet on adiposity were examined. MATERIALS/METHODS: 3T3-L1 cells were treated with arctiin (12.5 to $100{\mu}M$) during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining and intracellular triglyceride contents. The expressions of genes related to adipogenesis were measured by real-time RT-PCR and Western blot analyses. For in vivo study, C57BL/6J mice were first fed either a control diet (CON) or high-fat diet (HF) to induce obesity, and then fed CON, HF, or HF with 500 mg/kg BW arctiin (HF + AC) for four weeks. RESULTS: Arctiin treatment to 3T3-L1 pre-adipocytes markedly decreased adipogenesis in a dose-dependent manner. The arctiin treatment significantly decreased the protein levels of the key adipogenic regulators $PPAR{\gamma}$ and $C/EBP{\alpha}$, and also significantly inhibited the expression of SREBP-1c, fatty acid synthase, fatty acid-binding protein and lipoprotein lipase. Also, arctiin greatly increased the phosphorylation of AMP-activated protein kinase (AMPK) and its downstream target phosphorylated-acetyl CoA carboxylase. Furthermore, administration of arctiin significantly decreased the body weight in obese mice fed with the high-fat diet. The epididymal, perirenal or total visceral adipose tissue weights of mice were all significantly lower in the HF + AC than in the HF. Arctiin administration also decreased the sizes of lipid droplets in the epididymal adipose tissue. CONCLUSIONS: Arctiin inhibited adipogenesis in 3T3-L1 adipocytes through the inhibition of $PPAR{\gamma}$ and $C/EBP{\alpha}$ and the activation of AMPK signaling pathways. These findings suggest that arctiin has a potential benefit in preventing obesity.
This study was carried out to examine in vitro first whether the storage proteins, which the fat bodies of last larvae from Hyphantria cunea secrete into haemolymph, can be uptaked by the fat body cells of prepupa and then how the uptaked storage proteins can be accumulated in the fat body cells, if uptaken. The fat bodies which had been isolated from last instar larvae were cultured in 1 ml of Grace's insect medium containing $50{\mu}l$ of $^{3}H$-leucine (5.0 mCi/mol, Dupont) at $28{\pm}2^{\circ}C$ for 6 hrs. After the homogenates of the cultured fat bodies were centrifuged at 10,000 rpm for 10 minutes, the proteins included in the supernatant were separated by polyacrylamide gel electrophoreses (non-SDS, 6%). The next treatment of the electrophoresed gel was followed by rinsing. A storage protein band of several bands in the rinsed gel was sliced off. With elution of sliced storage protein bands in Tris-glycine buffer, the purification of radioactive storage proteins from fat bodies was finished. After the purified radioactive storage proteins were added in Grace's insect midis containing fat bodies of the prepupae, they were cultured for the randomly following minutes given as 3, 5, 7, 10, 15, 20 and 30 and for the randomly following hours given as 1, 2, 3 and 4 respectively. The double fixations of the cultured fat bodies in aldehyde and $OsO_4$, were followed by preparation of ultrathin sections from Epon-Araldite blocks through dehydration and embedding. The electron microscope autoradiographic treatment of all prepared sections were performed by the dipping method (Kim et al., 1987). The finally prepared specimens were examined with electron microscope. The fat body cells of the prepupa could be found to uptake the storage preteins of the last instar larvae, which were included in the culture medium, mostly by formation of coated vesicles. The in vitro uptake of the storage proteins actively occurred by 30 minutes after the addition of purified storage proteins in the culture medium. After culture for 7 minutes with the storage proteins, the uptaked radioactive storage proteins labelled a number of lysosomal granules. After culture for 20 minutes with the storage proteins, the radioactive storage proteins were finally incorporated and accumulated in lipid droplets and protein granules. The frequency in the fat body cell of radiolabelled lipid droplets occurs approximately 60%, while the frequency, in which the radiolabelled protein granules occurs in a fat body cell, is approximately 40%.
Kim, Jeong-Ja;Seo, Bu-Il;Choi, Hong-Sik;Kim, Seong-Mo;Woo, Chang-Hoon;Koo, Jin-Suk;Park, Gyu-Ryeol
Korean Journal of Oriental Medicine
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v.16
no.2
/
pp.167-179
/
2010
Objective : The present study has been undertaken to investigate the preventive effects of Daekumeumja on fatty degeneration of liver and immunosuppression induced by alcohol in rats. Method : Except for the normal group, we fed rat on 25% alcohol for 55 days. And Daekumeumja(DK) extract was administrated for the same period. We measured the serum component in rat's blood, weight of internal organs, liver triglyceride contents, histomorphometry and histopathological observation of internal organs. Results : 1. In the change measurement of serum components, DK group(50 mg/kg, 200 mg/kg) showed significant decrease of AST, ALT, albumin, ALP and triglyceride in comparison with those of the alcohol control group. 2. In the change measurement of internal organ's weight, DK group(50 mg/kg, 200 mg/kg) showed significant increase of relative body weights of liver, thymus and spleen in comparison with those of the alcohol control group. 3. DK group(50 mg/kg, 200 mg/kg) showed significant decrease of hepatic triglyceride contents in comparison with those of the alcohol control group. 4. In histomorphometrical changes of liver, DK group(50 mg/kg, 200 mg/kg) showed significant decrease of numbers of hepatocytes occupied by over 10% lipid droplets, percentages of regions occupied by lipid droplets and mean diameters of hepatocytes in comparison with those of the alcohol control group. In histomorphometrical changes of thymus, DK group(50 mg/kg, 200 mg/kg) showed significant increase of lobular thickness and cortex thickness in comparison with those of the alcohol control group. In histomorphometrical changes of spleen, DK group(50 mg/kg, 200 mg/kg) showed significant increase of splenic thicknesses, numbers of white pulps and mean diameters of white pulps in comparison with those of the alcohol control group. 5. In histopathological changes of liver, DK group(50 mg/kg, 200 mg/kg) showed effective inhibition of severe fatty changes in comparison with those of the alcohol control group. In histopathological changes of thymus and spleen, DK group(50 mg/kg, 200 mg/kg) showed effective inhibition of atrophic changes in comparison with those of the alcohol control group. Conclusions : Reviewing these experimental results, it appears that Daekumeumja have pharmaceutical preventive efficacy on fatty degeneration of liver and immunosuppression induced by alcohol in rats.
A study has been made to correlate morphological and biochemical differentiation in the oocytes of a tubiculous polychaete, Schizobranchia insignis. The pressent paper is concerned with an examination of the cytological changes during oogenesis and annual size distribution of oocytes, The oocytes are released from the ovary into the coelomic sac at the end of the oogonial division and grow to a maximum size (180 $\\mu$ diameter). Oogensis takes place continuously throughout a year, although the breeding season is the period between January and March. When the oocytes reach the largest size class, they remain constant in size thereafter and accumulate in the coelomic sac. The nucleolus, which first appears in the oocytes 5-10$\\mu$ diameter, grows in the early stages of oogenesis, becomes maximum in the oocytes 100-120$\\mu$ diameter, and is constant throughout the rest of the pertiod. The nuclelus initially has a single comartment but becomes bipartite prior to vitello genesis. Three types of yolk including lipid droplets, proteid granules and oval granules oof unknown composition form at different times of oogenesis. The lipid droplets and oval granules appear in the early stage, but mainly in the oocytes larger than 80 $\\mu$ diameter. Proteid yolk and cortical granules appear only in the oocytes larger than 80 $\\mu$ diameter Microvilli are abundant in the oocyte 80 $\\mu$ diameter and embedded in the vitelline membranc. In the oocytes 180 $\\mu$ diameter they have retracted from the vitelline membrane.
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