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Objective: To establish an efficacious and efficient fermentation method of enhancing the anti-adipogenesis effect of mulberry (Morus alba) leaves using Cordyceps militais. Methods: Dried mulberry leaves, dried mulberry leaves with 50% raw silkworm pupa and raw silkworm pupa were fermented with Cordyceps militais for 4 weeks at 25 °C, after which the dried mulberry leaves and fermented product were extracted with 70% ethanol and subjected to high performance liquid chromatography (HPLC). The contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid were determined. We then used the 3T3-L1 cells to investigate whether extracts of fermentation enhanced anti-adipogenesis activity in vitro. Results: HPLC showed that fermentation changed the contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid. Furthermore, fermented dried mulberry leaves with 50% raw silkworm pupa had a better efficacy of anti-adipogenesis than dried mulberry leaves, fermented dried mulberry leaves and fermented silkworm pupa and inhibited triglycerides accumulation and glucose consumption. Additionally, fermented dried mulberry leaves with 50% raw silkworm pupa inhibited PPAR-? signaling. Conclusions: Fermentation with Cordyceps militaris enhanced anti-adipogenesis efficacy of mulberry leaves.
ABSTRACT
Objective: To establish an efficacious and efficient fermentation method of enhancing the anti-adipogenesis effect of mulberry (Morus alba) leaves using Cordyceps militais. Methods: Dried mulberry leaves, dried mulberry leaves with 50% raw silkworm pupa and raw silkworm pupa were fermented with Cordyceps militais for 4 weeks at 25 °C, after which the dried mulberry leaves and fermented product were extracted with 70% ethanol and subjected to high performance liquid chromatography (HPLC). The contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid were determined. We then used the 3T3-L1 cells to investigate whether extracts of fermentation enhanced anti-adipogenesis activity in vitro. Results: HPLC showed that fermentation changed the contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid. Furthermore, fermented dried mulberry leaves with 50% raw silkworm pupa had a better efficacy of anti-adipogenesis than dried mulberry leaves, fermented dried mulberry leaves and fermented silkworm pupa and inhibited triglycerides accumulation and glucose consumption. Additionally, fermented dried mulberry leaves with 50% raw silkworm pupa inhibited PPAR-? signaling. Conclusions: Fermentation with Cordyceps militaris enhanced anti-adipogenesis efficacy of mulberry leaves.
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PURPOSE: Ascorbic acid has been reported to have an adipogenic effect on 3T3-L1 preadipocytes, while evidence also suggests that ascorbic acid reduces body weight in humans. In this study, we tested the effects of ascorbic acid on adipogenesis and the balance of lipid accumulation in ovariectomized rats, in addition to long-term culture of differentiated 3T3-L1 adipocytes. MATERIALS AND METHODS: Murine 3T3-L1 fibroblasts and ovariectomized rats were treated with ascorbic acid at various time points. In vitro adipogenesis was analyzed by Oil Red O staining, and in vivo body fat was measured by a body composition analyzer using nuclear magnetic resonance. RESULTS: When ascorbic acid was applied during an early time point in 3T3-L1 preadipocyte differentiation and after bilateral ovariectomy (OVX) in rats, adipogenesis and fat mass gain significantly increased, respectively. However, lipid accumulation in well-differentiated 3T3-L1 adipocytes showed a significant reduction when ascorbic acid was applied after differentiation (10 days after induction). Also, oral ascorbic acid administration 4 weeks after OVX in rats significantly reduced both body weight and subcutaneous fat layer. In comparison to the results of ascorbic acid, which is a well-known cofactor for an enzyme of collagen synthesis, and the antioxidant ramalin, a potent antioxidant but not a cofactor, showed only a lipolytic effect in well-differentiated 3T3-L1 adipocytes, not an adipogenic effect. CONCLUSION: Taking these results into account, we concluded that ascorbic acid has both an adipogenic effect as a cofactor of an enzymatic process and a lipolytic effect as an antioxidant.
Subject(s)
Animals , Female , Mice , 3T3-L1 Cells , Adipocytes/drug effects , Adipocytes/metabolism , Adipogenesis/drug effects , Antioxidants/pharmacology , Ascorbic Acid/pharmacology , Body Composition/drug effects , Body Weight/drug effects , Cell Differentiation/drug effects , Fibroblasts/drug effects , Fibroblasts/metabolism , Lipolysis/drug effects , Ovariectomy , Rats, Sprague-DawleyABSTRACT
Objective: To show the preventive effect of Brazilian propolis on metabolic syndrome.Methods: Nine Brazilian propolis were examined for inhibition ofα-glucosidase, absorption of sugar in mice, and lipid accumulation, glycerol 3-phosphate dehydrogenase, and adiponection production in mouse 3T3-L1 cells.Results: In nine Brazilian propolis, AF-06, AF-19, and AFG-06 propolis inhibited rat internal α-glucosidase, and AF-06 propolis inhibited the absorption of sugar in mice. In 3T3-L1 cells, AF-06 and AF-08 propolis inhibited accumulation lipid, and inhibited glycerol 3-phosphate dehydrogenase.Conclusion: Brazilian propolis AF-06 and AF-08 are natural products which offer promise in the prevention of diabetes and metabolic syndrome. Incorporating dietary supplements into a treatment plan with medicines with similar effects requires further study.
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Objective To investigate the influence of risperidone on differentiation of 3T3‐L1 pre‐adipocytes .Methods 3T3‐L1 pre‐adipocytes were induced to differentiate into mature adipocytes by adopting the classic hormone cocktail method and observed by the oil red O staining .Meanwhile ,the inducing medium was added with risperidone for studying its influence on 3T3‐L1 pre‐adi‐pocytes differentiation .Results 3T3‐L1 pre‐adipocytes were successfully differentiated into the mature adipocytes ,0 .1 ,1 ,10μmol/L risperidone all could inhibit the differentiation of 3T3‐L1 pre‐adipocytes .Conclusion Risperidone can inhibit the differentiation of 3T3‐L1 pre‐adipocytes .
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The dietary intake of whole grains is known to reduce the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. To investigate whether there are anti-adipogenic activities in various Korean cereals, we assessed water extracts of nine cereals. The results showed that treatment of 3T3-L1 adipocytes with Sorghum bicolor L. Moench, Setaria italica Beauvois, or Panicum miliaceum L. extract significantly inhibited adipocyte differentiation, as determined by measuring oil red-O staining, triglyceride accumulation, and glycerol 3-phosphate dehydrogenase activity. Among the nine cereals, P. miliaceum L. showed the highest anti-adipogenic activity. The effects of P. miliaceum L. on mRNA expression of peroxisome proliferator-activated receptor-gamma, sterol regulatory element-binding protein 1, and the CCAAT/enhancer binding protein-alpha were evaluated, revealing that the extract significantly decreased the expression of these genes in a dose-dependent manner. Moreover, P. miliaceum L. extract changed the ratio of monounsaturated fatty acids to saturated fatty acids in adipocytes, which is related to biological activity and cell characteristics. These results suggest that some cereals efficiently suppress adipogenesis in 3T3-L1 adipocytes. In particular, the effect of P. miliaceum L. on adipocyte differentiation is associated with the downregulation of adipogenic genes and fatty acid accumulation in adipocytes.
Subject(s)
3T3-L1 Cells , Adipocytes , Adipogenesis , Cardiovascular Diseases , Edible Grain , Chronic Disease , Down-Regulation , Fatty Acids , Fatty Acids, Monounsaturated , Glycerol , Glycerophosphates , Incidence , Obesity , Oxidoreductases , Panicum , Peroxisomes , RNA, Messenger , Setaria Plant , Sorghum , Sterol Regulatory Element Binding Protein 1 , Transcription Factors , WaterABSTRACT
Obesity and its related metabolic diseases become major health problems in the world.Adipose tissue plays an important role in the development of obesity.FSP27,a member of the CIDE family proteins,is expressed at high levels in white adipose tissue and differentiated 3T3L1 cells.The objective of current study is to establish a FSP27 knockdown preadipocyte cell line to investigate the in vivo function of mouse FSP27.The double strand siRNA of mouse FSP27 corresponding to nucleotides 270 to 291 was synthesized and inserted into pSilencer2.1.pSilencer-siFSP27 was co-transfected into 293T cells with the HA-mFSP27 expression vector to test its knock-down efficiency.The FSP27 siRNA was then transferred to a lentiviral vector.Lentivirus were generated and used to infect 3T3-L1 cells.It was shown here that lentivirus containing FSP27siRNA can effectively knockdown FSP27 expression in 3T3-L1 cells.Establishment of FSP27 knock-down cell line provides a useful tool for the study of in vivo function FSP27.
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The purpose of this study was to find out effects of treatment of ginsenoside Re, Rc and EGCG on mRNA expressions of leptin, hormone sensitive lipase (HSL) and resistin in 3T3-L1 adipocytes. The concentrations of EGCG were treated with 0.01 x 10(-7), 0.1 x 10(-7), 1 x 10(-7) and 1 x 10(-6) M or 100 microgram/ml ginsenoside Re, Rc in culture cell for 13 days. mRNA expression of leptin wasn't expressed in preadipocyte but according to differentiation of adipocyte, the that of mRNA expression was decreased at gensenosids or EGCG treated cells compared with non treated adipocyte. Expression of HSL mRNA was increased in G-Re, G-Rc and EGCG treated cells compared with non treated cells. The resistin level was significantly decreased in adipocytes treated with G-Re, G-Rc and EGCG. These pattern was similar to leptin expression.These results support that treatment of gensenosides or EGCG in 3T3-L1 adipocyte resulted to affect of leptin and resistin as well as HSL mRNA levels, accordingly, levels of leptin and HSL will be acted by signalling body fat stores to the hypothalamus which in turn regulates food intake and energy expenditure to maintain body weight homeostasis. And also regulation of resistin mRNA will prevent to diabetics attacked with obesity. In conclusion, we suggest that consumption of ginseng saponine or EGCG might prevent human diabetics or/and obesity.
Subject(s)
Humans , 3T3-L1 Cells , Adipocytes , Adipose Tissue , Body Weight , Catechin , Eating , Energy Metabolism , Homeostasis , Hypothalamus , Leptin , Obesity , Panax , Resistin , RNA, Messenger , Saponins , Sterol Esterase , TeaABSTRACT
Objective To establish the cell line stably expressing INSIG2 and observe its effecet on fat metabolism after overexpression of INSIG2.Methods The eukaryotic expression plasmid pcDNA3.1(+)-INSIG2 was constructed,which was transfected into 3T3-L1 cells.The expression of INSIG2 and related genes were detected by RT-PCR and immunohistochemistry,the contents of FFA in cell culture medium and adipocyte differentiation were detected by ELISA and Oil Red "O"staining respectively.Results After pcDNA3.1(+)-INSIG2 was transfected into the 3T3-L1 cells,the expression of INSIG1 mRNA and FAS mRNA were down-regulated,the content of FFA in the cell culture medium was decreased and adipocyte differentiation was drepressed.Conclusion The cell line stably expressing INSIG2 was successfully established,the transfected INSIG2 may have a drepressant effect on fat metabolism.
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Objective:To investigate the influence of high expression of Retn gene on glucose uptake by 3T3-L1 cells and to study its mechanism in inducing insulin resistance.Methods:(1)Radioimmunoassay was used to determine the glucose uptake by 3T3-L1 cells with low-,normal-and high-level Retn expression under basal and insulin-stimulated states.(2)RT-PCR and real-time RT-PCR were used to determine the mRNA levels of several glucose transport proteins in 3T3-L1 cells with different expression of Retn,including insulin receptor substrate-1(IRS-1),phosphatidylinositol 3-kinase(PI-3K),AKT-2,glucose transporter-4(GLUT-4),p38 mitogen-activated protein kinase(p38MAPK),and glycogen synthase kinase-3?(GSK-3?).(Results:) The uptake of glucose decreased with the increase of Retn expression under basal and insulin-stimulated conditions.The mRNA expression of 2 signal protein PI-3K and AKT-2 decreased with the increase of Retn expression;and the expression of GSK-3? and p38MAPK increased with the increase of Retn expression.Conclusion: Resistin protein can induce insulin resistance in adipocytes,which might be related to the expression changes of some proteins in PI-3K and Ras pathways.
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Expression of apelin mRNA in the isolated mouse adipocytes was confirmed by Northern blot. The level of apelin mRNA increased during stage of differentiation of 3T3-L1 cells. Insulin up-regulated the apelin expression in 3T3-L1 adipocytes, suggesting that apelin expression in adipocyte may correlate with obesity, insulin resistance and hypertension.