Expression Of Adipocyte-specific Markers Research Articles

319-LB: GLP-1 Receptor Agonist Promotes Brown Remodeling in? Human Adipose-Derived Stromal Cells Isolated from Omentum through Foxc2

Background: Brown adipose tissue (BAT) can disperse stored energy as heat. Promoting BAT-like features in white adipose tissue (WAT) is an attractive therapeutic approach to oppose obesity, glucose intolerance and insulin resistance. Accumulating evidence has revealed the significant role of glucagon-like peptide-1 (GLP-1) in improving insulin resistance in adipose tissue by reducing fat mass and adipocyte hypertrophy. Forkhead box C2 (FOXC2) plays a vital role in the regulation of lipid metabolism. Objects: Here, we investigated the effect of brown remodeling induced by the GLP-1 receptor (GLP-1R) agonist exenatide (exendin-4) in human adipose-derived stromal cells (hADSCs) isolated from omentum. We also looked at the role of FOXC2 in this process. Methods: hADSCs were isolated, cultured and differentiated as previously described. The effect of Exendin-4 on brown remodeling during differentiation of hADSCs was examined. The knockdown by siRNA was performed to determine the roles of FOXC2 in brown remodeling during differentiation of hADSCs. Results: In our study, oil red O test showed that Exendin-4 significantly increased the number of smaller lipid droplets, which is more similar to the morphological characteristics of brown fat. QPCR showed that Exendin-4 treatment could significantly up-regulate mRNA levels of some pro-brown adipogenic genes (UCP-1, FOXC2, PPARGC1A, PRDM16 and DIO2). Western blot also indicated that Exendin-4 could increase the protein levels of FOXC2 and UCP-1. Exendin-4-induced responses were partially blocked by GLP-1 receptor antagonist exendin9-39. The pro-brown adipogenic effects and increased expression of adipocyte specific markers caused by Exendin-4 was significantly reversed by the knockdown of FOXC2. Conclusions: Our data highlight that a GLP-1R agonist promotes brown remodeling of WAT in a FOXC2-dependent manner, this might be one of the mechanisms underlying its effect on weight loss. Disclosure X. He: None. L. Liu: None. W. Ke: None. Y. Li: None.

Increased Angiogenic and Adipogenic Differentiation Potentials in Adipose-Derived Stromal Cells from Thigh Subcutaneous Adipose Depots Compared with Cells from the Abdomen.

Adipose-derived stromal cells (ADSCs) may play a pivotal role by differentiating into multilineage cells or by secreting growth factors or cytokines in cell-assisted lipotransfer, which participates in adipose tissue regeneration. The angiogenic potential of various ADSCs from different anatomical regions remains uncertain. The authors sought to offer appropriate choices of sources of adipose-derived stromal cells for cell-assisted lipotransfer and tissue engineering. ADSCs were harvested from subcutaneous adipose depots in the abdomen and thighs. The expression of adipocyte-specific markers was evaluated, and Oil Red O staining was performed to assess the capacity for adipogenic differentiation. Angiogenic differentiation potential was evaluated by detecting the expression of vascular endothelial growth factor, vascular endothelial growth factor 2, and CD31. A tube formation assay was also performed to analyze the angiogenic differentiation capacity. ADSCs from the thigh showed more significant angiogenic and adipogenic potential. More lipogenesis was identified in ADSCs from the thigh, and this was accompanied by the enhancement of adipocyte markers. Angiogenesis was more vigorous in the thigh-derived stromal cells, and ADSCs from the thigh depot showed more junctions and longer tubule formation on Matrigel in vitro. Thigh-derived ADSCs exhibited greater capacity for adipogenic and angiogenic differentiation and would be a better option for cell-assisted lipotransfer and tissue engineering.

Long non-coding RNA TCONS_00041960 enhances osteogenesis and inhibits adipogenesis of rat bone marrow mesenchymal stem cell by targeting miR-204-5p and miR-125a-3p.

A growing number of long non‐coding RNAs (lncRNAs) have been found to be involved in diverse biological processes such as cell cycle regulation, embryonic development, and cell differentiation. However, limited knowledge is available concerning the underlying mechanisms of lncRNA functions. In this study, we found down‐regulation of TCONS_00041960 during adipogenic and osteogenic differentiation of glucocorticoid‐treated bone marrow mesenchymal stem cells (BMSCs). Furthermore, up‐regulation of TCONS_00041960 promoted expression of osteogenic genes Runx2, osterix, and osteocalcin, and anti‐adipogenic gene glucocorticoid‐induced leucine zipper (GILZ). Conversely, expression of adipocyte‐specific markers was decreased in the presence of over‐expressed TCONS_00041960. Mechanistically, we determined that TCONS_00041960 as a competing endogenous RNA interacted with miR‐204‐5p and miR‐125a‐3p to regulate Runx2 and GILZ, respectively. Overall, we identified a new TCONS_00041960‐miR‐204‐5p/miR‐125a‐3p‐Runx2/GILZ axis involved in regulation of adipogenic and osteogenic differentiation of glucocorticoid‐treated BMSCs.

Expression Profiles and Biological Roles of miR-196a in Swine.

MicroRNAs (miRNAs) are a class of small non-coding RNA molecules, which play important roles in animals by targeting mRNA transcripts for translational repression. Recent studies have demonstrated that miRNAs are involved in regulation of adipocyte development. The expression of miR-196a in different porcine tissues and developing fat tissues was detected, and gene ontology (GO) term enrichment was then used to predict the expression profiles and potential biological roles of miR-196a in swine. To further verify the roles of miR-196a in porcine adipocyte development, a recombinant adenovirus encoding miR-196a gene (Ad-miR-196a) was constructed and used to study the effect of miR-196a on preadipocyte proliferation and differentiation. Here, our data demonstrate that miR-196a displays a tissue-specific expression pattern and has comprehensive biological roles in swine, especially in adipose development. In addition, overexpression of miR-196a had no effect on preadipocyte proliferation, but induced preadipocyte differentiation by increasing expression of adipocyte specific markers, lipid accumulation and triglyceride content. These data represent the first demonstration of miR-196a expression profiles and roles in swine, thereby providing valuable insight into the functions of miR-196a in adipocyte biology.

Effect of Glucagon-like Peptide-1 on the Differentiation of Adipose-derived Stem Cells into Osteoblasts and Adipocytes.

ObjectivesGlucagon-like peptide-1 (GLP-1) is an intestinally secreted hormone and it plays an important role in the regulation of glucose homeostasis. However, the possible role of GLP-1 in the differentiation of adipose-derived stem cells (ADSCs) remains unknown. Therefore this study investigated the effect of GLP-1 on the differentiation of ADSCs into osteoblasts and adipocytes.MethodsADSCs were isolated from human adipose tissues of the abdomens, cultured and characterized by flow cytometry and multi-lineage potential assay. ADSCs were induced in osteogenic and adipogenic media treated with two different doses (10 and 100 nM) of GLP-1, and then the effect of GLP-1 on differentiation of ADSCs into osteoblast and adipocyte was examined. The signaling pathway involved in these processes was also examined.ResultsIsolated human ADSCs expressed mesenchymal stem cell (MSC) specific markers as well as GLP-1 receptor (GLP-1R) proteins. They also showed multiple-lineage potential of MSC. GLP-1 was upregulated the activity and mRNA expression of osteoblast-specific marker, alkaline phosphatase and the mineralization of calcium. In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2). This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).ConclusionThis result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation. The ERK signaling pathway seems to be involved in these differentiation processes mediated by GLP-1.

Control of adipogenesis by ezrin, radixin and moesin-dependent biomechanics remodeling

We have recently shown that altered stem cell biomechanics can regulate the lineage commitment through a family of the membrane–cytoskeleton linker proteins (ERM; ezrin, radixin, moesin). The ERM proteins not only modulate the cell stiffness and actin cytoskeleton organization, but also rearrange focal adhesions and therefore influence the biochemically-directed stem cell differentiation. Combining silencing RNA, atomic force microscopy, and fluorescence microscopy, the role of the ERM proteins involved in the regulation of stem cell biomechanics and adipogenic differentiation was quantitatively determined. Transient ERM knockdown by RNAi caused disassembly of actin stress fibers and focal adhesions and a decrease in the cell stiffness. The silencing RNA treatment not only induced mechanical changes in stem cells but impaired adipogenesis in a time-dependent manner. While siRNA ERM treatment at day 0 substantially interfered with adipogenesis, the same treatment at day 3 of adipogenic differentiation significantly facilitated adipogenesis, as assessed by the expression of adipocyte-specific markers. The intact biomechanics homeostasis appears to be critical for the adipogenic induction. These findings may lead to potential biomechanical intervention techniques and methodologies to control the fate and extent of adipogenesis that would likely be involved in stem cell-based therapeutics for soft tissue repair and regeneration.

Characterization of lipid metabolism in insulin-sensitive adipocytes differentiated from immortalized human mesenchymal stem cells

There is a great demand for cell models to study human adipocyte function. Here we describe the adipogenic differentiation of a telomerase-immortalized human mesenchymal stem cell line (hMSC-Tert) that maintains numerous features of terminally differentiated adipocytes even after prolonged withdrawal of the peroxisome proliferator activated receptor γ (PPARγ) agonist rosiglitazone. Differentiated hMSC-Tert developed the characteristic monolocular phenotype of mature adipocytes. The expression of adipocyte specific markers was highly increased during differentiation. Most importantly, the presence of the PPARγ agonist rosiglitazone was not required for the stable expression of lipoprotein lipase, adipocyte fatty acid binding protein and perilipin on mRNA and protein levels. Adiponectin expression was post-transcriptionally down-regulated in the absence of rosiglitazone. Insulin sensitivity as measured by insulin-induced phosphorylation of Akt and S6 ribosomal protein was also independent of rosiglitazone. In addition to commonly used adipogenic markers, we investigated further PPARγ-stimulated proteins with a role in lipid metabolism. We observed an increase of lipoprotein receptor (VLDLR, LRP1) and apolipoprotein E expression during differentiation. Despite this increased expression, the receptor-mediated endocytosis of lipoproteins was decreased in differentiated adipocytes, suggesting that these proteins may have an additional function in adipose tissue beyond lipoprotein uptake.

Association between altered expression of adipogenic factor SREBP1 in lipoatrophic adipose tissue from HIV-1-infected patients and abnormal adipocyte differentiation and insulin resistance

Lipodystrophy is a major side-effect of antiretroviral therapy but its pathophysiology remains elusive. In-vitro studies show that HIV-1-protease inhibitors affect adipocyte differentiation at an early step involving sterol-regulatory-element-binding-protein-1 (SREBP1), but in-vivo studies are lacking. We compared fat morphology and mRNA and protein expression of major adipocyte differentiation markers and cytokines in subcutaneous abdominal adipose tissue from 26 HIV-1-infected patients who developed peripheral lipoatrophy while on protease inhibitors and from 18 HIV-1-seronegative healthy controls. Patients' fat contained a higher proportion of small adipocytes than control fat, together with lower mRNA concentrations of the adipogenic differentiation factors CCAAT-enhancer binding protein (C/EBP) beta and alpha, peroxisome proliferator-activated receptor (PPAR) gamma, and the 1c isoform of SREBP1, with a median decrease of 93% in the latter. The SREBP1 protein concentration was increased 2.6-fold, whereas the PPARgamma protein concentration was decreased by 70%. The expression of adipocyte-specific markers, including leptin, was lower in fat from patients than in fat from controls, whereas expression of tumour necrosis factor (TNF) alpha was higher and correlated negatively with the expression of SREBP1c and downstream adipogenic factors. SREBP1c mRNA concentrations correlated negatively, and TNFalpha mRNA concentrations positively, with glycaemia and insulin resistance, but did not correlate with lipid variables. The altered differentiation status of peripheral adipocytes in HIV-1-infected patients with antiretroviral-induced lipoatrophy is associated with greatly reduced SREBP1c expression. Since the differentiation factor SREBP1 is rapidly targeted by protease inhibitors in vitro, our results suggest that SREBP1c could be an important mediator of peripheral lipoatrophy in this setting, leading to metabolic alterations such as insulin resistance.