Effect of Sake Lees on the Inhibition of Lipid Accumulation in Adipocytes

Recent lifestyle changes, such as the Westernization of diets and the rise in the prevalence of obesity, with an associated increase in the number of patients with lifestyle-related diseases, have become a social issue. Fermented food has attracted attention as a solution to problems caused by obesity. Sake lees, a byproduct of sake brewing, represent one such food that is expected to have health benefits. In this study, we investigated the effects of sake lees components on preadipocytes (3T3-L1). We cultured preadipocytes in a medium with indigestible sake lees components (ISLCs) to investigate lipid accumulation, analyzed the glycerol 3-phosphate dehydrogenase (GPDH) and LPL activities of those cells, and performed a real-time PCR analysis of the IL6 expression in the cells. The results show that lipid accumulation and GPDH activity were significantly decreased in adipocytes treated with 1.0 mg/mL ISLCs compared to untreated cells. Furthermore, the expression of IL6 in adipocytes treated with 1.0 mg/mL ISLCs was significantly decreased and the lipase activity was significantly increased in adipocytes treated with ISLCs after differentiation. IL6 is known to have multiple functions in adipose tissue. In conclusion, ISLCs were associated with reduced lipid accumulation in adipocytes, with effects on IL6 expression and LPL activity observed throughout the differentiation period.

Phytochemicals derived from soya bean husk exert hypoglycemic and anti-adipogenic properties in cell culture models

Purpose Literature has shown that phenolic acids and flavonoids are bearing with hypoglycemic and anti-adipogenic properties. Therefore, this study aims to evaluate the possibility of phenolic-rich soya bean husk powder extract (SHPE) in combating diabetes and obesity using in vitro models. Design/methodology/approach The hypoglycemic properties were evaluated by determining the ability of SHPE (25-100 µg/mL) in inhibiting a-amylase and a-glucosidase enzymes and in triggering insulin secretion in BRIN-BD11 cells. Murine 3T3-L1 adipocytes were used for evaluating the anti-adipogenic properties of SHPE through the determination of relative lipid accumulation, triglyceride content and glycerol-3-phosphate dehydrogenase (GPDH) activity. Findings The hypoglycemic properties of SHPE was in the dose-dependent manner, where 100 µg SHPE/mL exhibited a significant higher (p < 0.05) a-amylase inhibitory activity (56.8 ± 0.11 per cent) and insulin secretion activity (0.73 ± 0.02 µg/l) against other concentrations. In contrast to the aforementioned findings, a significant lower a-glucosidase inhibitory activity (52.0 ± 0.44 per cent) was also observed in 100 µg SHPE/mL. Nevertheless, findings revealed that all the SHPE were able to inhibit the activity of a-amylase and a-glucosidase and stimulated the insulin secretion in BRIN-BD11 cells. On the other hand, the anti-adipogenic properties of SHPE were in the reverse dose-dependent manner, where 100 µg SHPE/mL demonstrated a significant lower (p < 0.05) relative lipid accumulation (48.5 ± 0.03 per cent), intracellular triglyceride content (5.7 ± 0.07 mg/dL) and GPDH activity (1.0 ± 0.01 mU/mL). These findings reflected that 100 µg SHPE/mL was a potent anti-adipogenic agent when compared with other concentrations. In conclusion, soya husk could emerge as a potential hypoglycemic and anti-adipogenic agents in in vitro models. Originality/value This was the first study to explore the effectiveness of phytochemicals derived from soya bean husk in ameliorating hyperglycemia and adipogenesis. Promising findings that derived from the present study could enable the scientists to re-evaluate the potential use of agricultural wastes, especially in the formulation of nutraceuticals.

The Inhibitory Effects of Djulis (Chenopodium formosanum) and Its Bioactive Compounds on Adipogenesis in 3T3-L1 Adipocytes

The aim of this study was to provide new insights into the role of the ethanolic extracts of Djulis (Chenopodium formosanum, EECF) and its bioactive compounds in preventing adipogenesis in 3T3-L1 adipocytes. The results demonstrated EECF significantly inhibited oil red O-stained material (OROSM), triglyceride levels and glycerol-3-phosphate dehydrogenase (GPDH) activity in 3T3-L1 adipocytes. The expression of the critical molecules involved in lipid synthesis such as PPARγ, C/EBPα and SREBP-1c was attenuated in EECF-treated cells. According to HPLC-DAD and HPLC-MS/MS analysis, rutin, kaempferol, betanin and another nine compounds were present in EECF. The suppression of lipid accumulation by rutin, kaempferol and betanin occurred by decreasing the gene expression of PPARγ, C/EBPα and SREBP-1c. Taken together, these findings suggest the presence of bioactive compounds in EECF may partly account for the anti-adipogenesis of EECF and EECF is therefore a potentially lipid lowering functional food.

ZBTB16 Overexpression Enhances White Adipogenesis and Induces Brown-Like Adipocyte Formation of Bovine White Intramuscular Preadipocytes

Background/Aims: Our study aims to characterize functions of ZBTB16 gene in the process of intramuscular fat (IMF) deposition and metabolism of bovine, thereby providing insights into mechanisms for the use of ZBTB16 in fat management. Methods: Primary preadipocytes derived from bovine IMF tissue were isolated and used as the in vitro cell model. An adenovirus Ad-ZBTB16 was transfected into bovine preadipocytes to overexpress the ZBTB16 gene. By using real-time quantitative PCR (RT-qPCR), western blotting, Oil Red-O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity assay, and cell counting kit-8 (CCK-8) test, adipogenic and proliferative signals in adipocytes were monitored to investigate effects of ZBTB16 on adipogenesis of bovine preadipocytes. Results: After transfection, mRNA and protein levels of ZBTB16 gene were significantly increased. Enhanced ZBTB16 significantly promoted preadipocyte differentiation, as evidenced by accelerated lipid accumulation, enhanced GPDH activity, consistently increased mRNA expressions of adipogenic key transcription factors PPARγ, C/EBPα, FABP4, and ADIPOQ, and markedly increased protein expressions of PPARγ and FABP4. No difference was observed concerning proliferation of preadipocytes after treatment with Ad-ZBTB16. Furthermore, relative mRNA levels of brown adipocyte selective genes (PRDM16, UCP1, Cidea, Cox8b, and PGC-1α) and beige adipocyte selective genes (CD137, TMEM26, and Tbx1) as well as UCP1 protein expression were significantly increased by Ad-ZBTB16. Meanwhile, Ad-ZBTB16 treatment remarkably induced mitochondrial biogenesis and increased relative mitochondrial DNA (mtDNA) copy number in bovine adipocytes. Conclusion: These results suggest that ZBTB16 overexpression can promote white adipogenesis and induce brown-like adipocyte formation for bovine white intramuscular preadipocytes.

Cocktail supplement with rosiglitazone: a novel inducer for chicken preadipocyte differentiation in vitro

Chicken preadipocytes cultured in cocktail supplement with rosiglitazone resulted in a marked increase in lipid droplet accumulation, glycerol-3-phosphate dehydrogenase (GPDH) activity and mRNA expression of adipocyte fatty acid-binding protein (aP2), G0/G1 switch gene 2 (G0S2), peroxisome proliferator-activated receptor γ (PPARγ) and lipolysis. The present study provides a novel induction method for in vitro chicken preadipocyte differentiation.

Traditional Herbal Formula Oyaksungi-San Inhibits Adipogenesis in 3T3-L1 Adipocytes

Background. Oyaksungi-san (OYSGS) is a herbal formula that has been used for treating cardiovascular diseases in traditional Asian medicine. Here, we investigated the antiadipogenic effect of OYSGS extract in 3T3-L1 adipose cells.Methods. 3T3-L1 preadipocytes were differentiated into adipocytes with or without OYSGS. After differentiation, we measured Oil Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, leptin production, mRNA, and protein levels of adipogenesis-related factors.Results. OYSGS extract dramatically inhibited intracellular lipid accumulation in the differentiated adipocytes. It also significantly suppressed the (GPDH) activity, triglyceride (TG) content, and leptin production by reducing the expression of adipogenesis-related genes including lipoprotein lipase, fatty acid binding protein 4, CCAAT/enhancer-binding protein-alpha (C/EBP-α), and peroxisome proliferator-activated receptor gamma (PPAR-γ). Furthermore, OYSGS clearly enhanced phosphorylation of AMP-activated protein kinase (AMPK) as well as its substrate acetyl CoA (ACC) carboxylase.Conclusions. Our results demonstrate that OYSGS negatively controls TG accumulation in 3T3-L1 adipocytes. We suggest antiadipogenic activity of OYSGS and its potential benefit in preventing obesity.

Effect of Collagen Coating on Lipid Accumulation and Differentiation on 3T3-L1 Preadipocyte Treated with Conjugated Linoleic Acid

The purpose of this study is to make use of trans10,cis12 CLA (t-CLA) that has potential for proliferation and differentiation to form adipocyte on the collagen-coated surface. Results provided evidences of good adhesion, growth, viability, and differentiation of adipocyte on collagen-coated surface compared with non-coated surface. Also, the results showed that mouse 3T3-L1 preadipocyte can be successfully and reproducibly cultured on the collagen-coated surface, and the adipocyte precursor cells placed on the collagen-coated surface are able to undergo full maturation into adipocytes in the control cells. Moreover, glycerol-3-phosphate dehydrogenase (GPDH) activity in 3T3-L1 preadipocyte cultured on collagen-coated surface with t-CLA was higher than that on polystyrene (PS) surface due to higher cell adhesion and cell viability. These results suggest that collagen coating may provide a promising approach to develop a new adipocyte replacement strategy using CLA.

Role of intracellular calcium in human adipocyte differentiation

Shi, Hang, Yuan-Di Halvorsen, Pamela N. Ellis, William O. Wilkison, and Michael B. Zemel. Role of intracellular calcium in human adipocyte differentiation. Physiol Genomics 3: 75–82, 2000.—Intracellular calcium ([Ca2+]i) modulates adipocyte lipid metabolism and inhibits the early stages of murine adipogenesis. Consequently, we evaluated effects of increasing [Ca2+]i in early and late stages of human adipocyte differentiation. Increasing [Ca2+]i with either thapsigargin or A23187 at 0–1 h of differentiation markedly suppressed differentiation, with a 40–70% decrease in triglyceride accumulation and glycerol-3 phosphate dehydrogenase (GPDH) activity ( P < 0.005). However, a 1-h pulse of either agent at 47–48 h only modestly inhibited differentiation. Sustained, mild stimulation of Ca2+ influx with either agouti protein or 10 mM KCl-induced depolarization during 0–48 h of differentiation inhibited triglyceride accumulation and GPDH activity by 20–70% ( P < 0.05) and markedly suppressed peroxisome proliferator-activated receptor gamma (PPARγ) expression. These effects were reversed by Ca2+ channel antagonism. In contrast, Ca2+ pulses late in differentiation (71–72 h or 48–72 h) markedly increased these markers of differentiation. Thus increasing [Ca2+]i appears to exert a biphasic regulatory role in human adipocyte differentiation, inhibiting the early stages while promoting the late stage of differentiation and lipid filling.

Dehydroepiandrosterone attenuates preadipocyte growth in primary cultures of stromal-vascular cells

The purpose of this study was to determine whether the antiobesity actions of dehydroepiandrosterone (DHEA) are due to an influence on preadipocyte proliferation and/or differentiation in primary cultures of pig and rat stromal-vascular (SV) cells. Pig SV cells were isolated from dorsal subcutaneous adipose tissue of 7-day-old pigs. For the proliferation assays, pig SV cells were grown for 4 days in plating medium containing DHEA at 0, 15, 50, or 150 μM. For the differentiation assays, pig SV cells were grown in plating medium for 3 days and then switched to a serum-free medium containing DHEA at 0, 15, 50, or 150 μM for the next 6 days. Rat SV cells were isolated from inguinal fat pads of 5-wk-old male rats. Rat SV cells were exposed to DHEA at 0, 5, 25, or 75 μM during proliferation. For the differentiation assays, rat SV cells were grown for 8 days in a serum-free medium containing DHEA at 0, 5, 25, or 75 μM. Preadipocyte differentiation [lipid staining, glycerol-3-phosphate dehydrogenase (GPDH) activity] and proliferation (preadipocyte-specific antigen staining) decreased with increasing levels of DHEA in cultures of pig SV cells. In cultures of rat SV cells, preadipocyte differentiation (lipid staining, GPDH activity) and proliferation ([3H]thymidine incorporation) were decreased in the 25 and 75 μM DHEA groups compared with the control and 5 μM DHEA groups. The level of expression of CCAAT enhancer binding protein-α, a master regulator of adipogenesis, in cultures of pig SV cells treated with 150 μM DHEA was 38% of control cultures. These data support the hypothesis that DHEA directly attenuates adipogenesis via attenuation of preadipocyte proliferation and differentiation.

Development of intramuscular fat in Wagyu beef cattle depends on adipogenic or antiadipogenic substances present in serum

In blood, there are many kinds of adipogenic or antiadipogenic factors such as hormones and vitamins. In this study, adipogenic activity in sera of fattened beef cattle was evaluated using cultured mouse 3T3-L1 preadipocytes. After the preadipocytes were grown to reach confluence, serum of fattened beef cattle was added into the culture medium (10%, vol/vol)for 3 days, and thereafter cellular sn-glycerol-3-phosphate dehydrogenase (GPDH) activity was determined as an index ofadipocyte differentiation. Sera were collected from 19 beef cattle (Wagyu and Wagyu × Holstein cross cattle) from three different farms at slaughter. Cellular GPDH activity was significantly different among the farms, and was affected by sex difference (i.e. sera from fattened heifers induced higher GPDH activity than those from steers). There was a positive correlation between GPDH activity and beef marbling performance (T = 0·62, P < 0·02), suggesting that serum factor(s) play a role in development of intramuscular fat deposition. Adipogenic activity was negatively correlated with serum retinol concentration (r = −0·73, P < 0·001). Neither serum cholesterol, triacylglycerol nor non-esterified fatty acid was related to adipogenic activity.Furthermore, serum retinol concentration was negatively correlated with beef marbling performance. These data imply that retinol level in blood during the fattening period may influence intramuscular fat deposition of beef cattle through its antiadipogenic action on preadipocytes present in muscle tissues.