Expression Levels Of Adipogenic Genes Research Articles

DUSP8-attenuated ERK1/2 signaling mediates lipogenesis and steroidogenesis in chicken granulosa cells

The lipogenesis and steroidogenesis of granulosa cells are crucial during follicular development, yet it remains unclear whether dual-specificity phosphatase 8 (DUSP8) is involved. In this study, the specific role of DUSP8 in lipogenesis and steroidogenesis was investigated through culturing chicken granulosa cells in vitro. The results revealed that the expression levels of adipogenic genes were elevated after DUSP8 overexpression and reduced after knockdown. The same was observed for lipid deposition in granulosa cells. Meanwhile, the steroidogenic gene expression and progesterone synthesis were promoted after DUSP8 overexpression and inhibited after knockdown. In addition, we also found that DUSP8 blocked the phosphorylation of extracellular regulatory kinase 1/2 (ERK1/2). Based on the previous results that activated ERK1/2 signaling inhibited lipid deposition and progesterone synthesis in chicken granulosa cells, we demonstrated that DUSP8 promoted lipid deposition and progesterone synthesis through mediating the ERK1/2 signaling pathway. The results will improve our understanding of the molecular regulatory mechanisms regarding lipid metabolism and progesterone synthesis in chicken granulosa cells.

Comparison of physicochemical properties of sorghum extract by ethanol concentration and its anti-adipogenic effect in 3T3-L1cells.

Sorghum is a vital cereal source that has various phenolic compounds and potential health-promoting benefits. This study evaluated the phenolic content, antioxidant and anti-obesity effects of sorghum extract (SE) prepared using three solvent systems: 50% (SE50), 80% (SE80), and 100% (SE100) ethanol. The results showed that SE50 exhibited the highest total polyphenol and flavonoid content among the sorghum extracts using different ethanol concentrations as extraction solvents. In addition, SE50 showed significantly higher antioxidant capacity than the other extracts. Interestingly, SE50 significantly inhibited lipid accumulation in 3T3-L1 adipocytes; however, SE80 and SE100 had no beneficial effects. Moreover, SE50 significantly downregulated the mRNA expression levels of adipogenic genes (Cebpα, Pparγ, and Fabp4) and lipogenic genes (Srebp1c, Fas, and Scd1). These results suggest that SE50 is superior to other ethanol extracts in phenolic contents, antioxidant and anti-obesity activities, and it could be used as a nutraceutical for anti-obesity.

Inhibition of PPARγ Gene Particle Downregulates the Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells into Adipocytes

This study aims to evaluate the effect of peroxisome proliferator-activated receptor (PPAR) γ gene inhibition on the adipogenic differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs). Primary BMSCs were isolated from rabbit bone marrow, cultured, and the markers of BMSCs on cell’s surface were analyzed using flow cytometry. The experiment involved five groups, namely, control: untreated BMSCs; model: BMSCs treated with ethanol; empty siRNA: BMSCs treated with ethanol + empty siRNA; PPARγ: BMSCs treated with ethanol + PPARγ siRNA; and PPARγ inhibitor: BMSCs treated with ethanol + T0070907. RT-PCR and Western blotting were used to detect changes in the expression level of PPARγ, PETALA2 (AP2), lipoprotein lipase (LPL), fatty acid transport protein (FATP) 1, and fatty acid transporter (FAT). Adipocyte count and triacylglycerol content of the model and the empty siRNA groups were considerably greater than the control group ( P < 0.01 ). After the inhibition with PPARγ or T0070907, adipocyte count and triacylglycerol content of the PPARγ and T0070907 groups were significantly reduced ( P < 0.01 ), with no statistically significantly difference than the control group ( P > 0.05 ). The expression levels of PPARγ gene and protein in the model and empty siRNA groups were ominously enhanced than the control group ( P < 0.01 ), and after inhibition with PPARγ or T0070907, the PPARγ gene or protein expression level of PPARγ and T0070907 groups significantly reduced ( P < 0.01 ), with no statistically significance difference compared to the control group ( P > 0.05 ). The expression levels of Ap2, LPL, FATP1, and FAT genes in the model and empty siRNA groups were considerably greater compared to the control group ( P < 0.01 ). Inhibition with PPARγ or T0070907 in the PPARγ and T0070907 groups, respectively, lead to significantly reduced expression levels of adipogenic genes ( P < 0.01 ), with no statistically significance difference compared to the control ( P > 0.05 ). Inhibition of PPARγ gene downregulates the differentiation of BMSCs into adipocytes, indicating its putative role in the expression of adipogenic genes.

Effects of puerarin on the AKT signaling pathway in bovine preadipocyte differentiation.

ObjectivePuerarin has the potential of regulating the differentiation of preadipocytes, but its mechanism of action has not yet been elucidated. Adipocytes found in adipose tissue, the main endocrine organ, are the main sites of lipid deposition, and are widely used as a cell model in the study of in vitro fat deposition. This study aimed to investigate the effects of puerarin on adipogenesis in vitro.MethodsPuerarin was added to the culture medium during the process of adipogenesis. The proliferation and differentiation of bovine preadipocytes was measured through cell viability and staining with oil red O. The content of triacylglycerol was measured using a triglyceride assay kit. The mRNA and protein expression levels of adipogenic genes, peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer-binding protein-α, were measured using quantitative real-time polymerase chain reaction and western blotting, respectively.ResultsThe addition of puerarin significantly increased adipogenesis of bovine preadipocytes and enhanced the mRNA and protein level expression of PPARγ (p<0.01). The expression of P-Akt increased after adipogenic hormonal induction, whereas puerarin significantly increased PPARγ expression by promoting the Akt signaling component, P-Akt. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473, which may activate the downstream signaling of the Akt pathway.ConclusionPuerarin was able to promote the differentiation of preadipocytes and improve fat deposition in cattle. The mechanism of adipogenesis was found to be related to the phosphorylation level of Ser473.

Adipocyte-specific DKO of Lkb1 and mTOR protects mice against HFD-induced obesity, but results in insulin resistance

Liver kinase B1 (Lkb1) and mammalian target of rapamycin (mTOR) are key regulators of energy metabolism and cell growth. We have previously reported that adipocyte-specific KO of Lkb1 or mTOR in mice results in distinct developmental and metabolic phenotypes. Here, we aimed to assess how genetic KO of both Lkb1 and mTOR affects adipose tissue development and function in energy homeostasis. We used Adiponectin-Cre to drive adipocyte-specific double KO (DKO) of Lkb1 and mTOR in mice. We performed indirect calorimetry, glucose and insulin tolerance tests, and gene expression assays on the DKO and WT mice. We found that DKO of Lkb1 and mTOR results in reductions of brown adipose tissue and inguinal white adipose tissue mass, but in increases of liver mass. Notably, the DKO mice developed fatty liver and insulin resistance, but displayed improved glucose tolerance after high-fat diet (HFD)-feeding. Interestingly, the DKO mice were protected from HFD-induced obesity due to their higher energy expenditure and lower expression levels of adipogenic genes (CCAAT/enhancer binding protein α and PPARγ) compared with WT mice. These results together indicate that, compared with Lkb1 or mTOR single KOs, Lkb1/mTOR DKO in adipocytes results in overlapping and distinct metabolic phenotypes, and mTOR KO largely overrides the effect of Lkb1 KO.

Altered Adipose-Derived Stem Cell Characteristics in Macrodactyly

Macrodactyly is a congenital disease characterized by aggressive overgrowth of adipose tissue in digits or limbs frequently accompanied with hyperostosis and nerve enlargement; its pathological mechanism is poorly understood. Adipose-derived stem cells (ASCs) have been extensively studied in tissue engineering and regenerative medicine as an ideal alternative substitute for bone marrow-derived mesenchymal stem cells (BM-MSCs), but their pathological role is largely unknown. In this study, ASCs from macrodactyly adipose tissues (Mac-ASCs) were isolated and compared to ASCs derived from the normal abdominal subcutaneous adipose tissue (Sat-ASCs) for cell morphology, surface marker expression, proliferation rate, and tri-lineage differentiation potential. Despite similar cell morphology and cell surface marker expression, Mac-ASCs showed higher cell proportion in the S phase and increased proliferation compared with Sat-ASCs. Moreover, osteogenic and chondrogenic differentiation capacities were enhanced in Mac-ASCs, with reduced adipogenic potential. In addition, the expression levels of adipogenic genes were lower in undifferentiated Mac-ASCs than in Sat-ASCs. These findings unraveled enhanced proliferation activity, a regression in the differentiation stage, and greater potentiality of ASCs in macrodactyly, which could contribute to hyperostosis and nerve enlargement in addition to adipose tissue overgrowth in patients.

Anti-oxidant and anti-adipogenic effects of acorn (Quercus acutissima CARR.) shell extracts via regulation of wnt signaling in 3T3-L1 cells

Acorn (Quercus acutissima CARR.) is a nut from the Fagaceae family that has been used in traditional medicine for many years. However, shells from acorns are regarded as a by-product and are mostly discarded. Anti-adipogenic activities of acorn shells were investigated using 3T3-L1 cells and methanol shell extracts (AE-M). AE-M demonstrated Cu2+-chelation activities and anti-oxidant activities via reduction of oxidative stress levels induced using AAPH. Six days after adipocyte differentiation, 50 and 100 μg/mL AE-M completely suppressed 3T3-L1 adipogenesis and the anti-adipogenic effect was stronger than for the positive control 50 μM quercetin. Treatment with AE-M in 3T3-L1 cells reduced mRNA expression levels of adipogenic genes. AE-M-inhibition was found in pre-adipogenic, early, and intermediate stages of adipogenesis in 3T3-L1 cells. The Wnt/β-catenin signaling pathway is required for AE-M-inhibition of 3T3-L1 adipogenesis.

Anti-adipogenic activity of blue mussel (Mytilus edulis) extract by regulation of 3T3-L1 adipogenesis through Wnt/β-catenin signaling pathway

The blue mussel (Mytilus edulis) is widely distributed in European waters but it is little consumed. Although several studies have been reported biological activities of mussels, the anti-adipogenic activity of blue mussel and the molecular mechanism in anti-obese activity remain unknown. Here, we prepared a water extract of blue mussel (BME) and its anti-adipogenic activity was assessed in 3T3-L1 cells. BME treatment dose-dependently inhibited 3T3-L1 lipid accumulation, as judged by Oil Red O staining and microscopic analysis. Consistent with this, BME treatment markedly suppressed mRNA expression levels of adipogenic genes, such as C/EBPβ, PPARγ, C/EBPα, and FAS, in 3T3-L1 mature adipocytes. BME-inhibited lipid accumulation in 3T3-L1 adipocytes was limited with treatment after adipocyte differentiation, particularly in the early and intermediate stages. We also showed that the Wnt/β-catenin signaling pathway is required in BME-inhibited 3T3-L1 adipogenesis. These finding may provide scientific evidence for the development of functional foods against obesity.

Impact of adipogenic differentiation on stemness and osteogenic gene expression in extensive culture of human adipose-derived stem cells.

IntroductionAdipose tissue is a source of multipotent adult stem cells. Most studies on human adipose-derived stem cells (ASC) have been on the early passages. Studies in extensive expansion have not been well established yet. In this study, we aim to investigate the effects of extensive expansion on the adipogenic differentiation capability of ASC.Material and methodsThe ability of ASC to undergo adipogenic differentiation in extensive expansion was evaluated by morphological changes, differentiation assay by using Oil Red O staining and changes in the genes expression levels of adipogenic genes, osteogenic genes and stemness genes using quantitative polymerase chain reaction (qPCR) after induction.ResultsMorphological study showed that the formation of lipid droplets can be observed at all passages but decreased at P20 after induction. Data from qPCR showed that most adipogenicgenes expression increased significantlyat P5, P10 and P15 but decreased at P20 after induction. On the other hand, osteogenic genes showed no significant changes after adipogenic induction indicating low potentiality of adipogenic-induced ASC to become osteogenic cells. While stemness genes expression levels showed a decrease or no significant changes after adipogenic induction except Nanog3, which showed a significant increase at P15 and P20.ConclusionsThe ability of ASC to differentiate into mature adipogenic cells decreased after P10 and the decrease in the osteogenics gene expression level during adipogenic induction suggested that the osteogenesis and adipogenesis are not parallel events.

Mechano growth factor E peptide regulates migration and differentiation of bone marrow mesenchymal stem cells

IGF1Ec in humans or IGF1Eb in rodents (known as mechano growth factor (MGF)) has a unique E domain, and the C-terminal end of the E domain (MGF E peptide) plays important roles in proliferation, migration and differentiation of many cell types. Bone marrow mesenchymal stem cells (BMSCs) have multiple differentiation potentials and are considered as perfect seed cells for tissue repair. But the role of MGF E peptide on BMSCs is seldom investigated and the mechanism is still unclear. In this study, we investigated the effects of MGF E peptide on rat BMSCs (rBMSCs). Our results revealed that treatment with MGF E peptide had no effect on BMSC proliferation. However, both wound-healing and transwell assays indicated that MGF E peptide could significantly enhance rBMSCs migration ability. Further analysis indicated that MGF E peptide also reduced the expression levels of osteogenic genes, but increased the expression levels of adipogenic genes. Analysis of molecular mechanism showed that phosphorylation-Erk1/2 was activated by MGF E peptide and blockage of either Erk1/2 or IGF1 receptor could repress the migration effect of MGF E peptide. In conclusion, MGF E peptide is able to inhibit osteogenic differentiation but promote adipogenic differentiation. In addition, the migration effect of MGF E peptide on rBMSCs depends on IGF1 receptor via Erk1/2 signal pathway.