RESUMEN
BACKGROUND: Obesity is closely associated with lipid accumulation and intestinal microbiota dysbiosis. It has been proved that probiotics supplement contributes to alleviate obesity. The objective of this study was to investigate the mechanism by which Lactobacillus plantarum HF02 (LP-HF02) alleviated lipid accumulation and intestinal microbiota dysbiosis in high-fat diet-induced obese mice. RESULTS: Our results showed that LP-HF02 ameliorated body weight, dyslipidemia, liver lipid accumulation, and liver injury in obese mice. As expected, LP-HF02 inhibited pancreatic lipase activity in small intestinal contents and increased fecal triglyceride levels, thereby reducing dietary fat hydrolysis and absorption. Moreover, LP-HF02 ameliorated the intestinal microbiota composition, as evidenced by the enhanced ratio of Bacteroides to Firmicutes, the decreased abundance of pathogenic bacteria (including Bacteroides, Alistipes, Blautia, and Colidextribacter) and the increased abundance of beneficial bacteria (including Muribaculaceae, Akkermansia, Faecalibaculum, and Rikenellaceae_RC9_gut_group). LP-HF02 also increased fecal short-chain fatty acids (SCFAs) levels and colonic mucosal thickness, and subsequently decreased serum lipopolysaccharide (LPS), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α) levels in obese mice. Additionally, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot results demonstrated that LP-HF02 ameliorated hepatic lipid accumulation via activating the adenosine monophosphate (AMP)-activated protein kinase (AMPK) pathway. CONCLUSION: Therefore, our results indicated that LP-HF02 could be considered as a probiotic preparation for preventing obesity. © 2023 Society of Chemical Industry.
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Disbiosis , Microbioma Gastrointestinal , Lactobacillus plantarum , Metabolismo de los Lípidos , Obesidad , Lactobacillus plantarum/química , Lactobacillus plantarum/metabolismo , Ratones Endogámicos C57BL , Masculino , Animales , Ratones , Disbiosis/complicaciones , Disbiosis/metabolismo , Dieta Alta en Grasa , Obesidad/complicaciones , Obesidad/metabolismo , Heces/química , Adipocitos/química , Adipocitos/metabolismoRESUMEN
Exposure to fine particulate matter (PM2.5 ) from incomplete fossil fuel combustion (coal, oil, gas and diesel) has been linked to increased morbidity and mortality due to metabolic diseases. PM2.5 exaggerate adipose inflammation and insulin resistance in mice with diet-induced obesity. Here, we elucidate the hypothesis that such systemic effects may be triggered by adhered particle components affecting adipose tissue directly. Studying adipocytes differentiated from primary human mesenchymal stem cells, we found that lipophilic organic chemicals (OC) from diesel exhaust particles induced inflammation-associated genes and increased secretion of the chemokine CXLC8/interleukin-8 as well as matrix metalloprotease 1. The oxidative stress response gene haem oxygenase-1 and tumour necrosis factor alpha were seemingly not affected, while aryl hydrocarbon receptor-regulated genes, cytochrome P450 1A1 (CYP1A1) and CYP1B1 and plasminogen activator inhibitor-2, were clearly up-regulated. Finally, expression of ß-adrenergic receptor, known to regulate adipocyte homoeostasis, was down-regulated by exposure to these lipophilic OC. Our results indicate that low concentrations of OC from combustion particles have the potential to modify expression of genes in adipocytes that may be linked to metabolic disease. Further studies on mechanisms linking PM exposure and metabolic diseases are warranted.
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Contaminantes Atmosféricos , Células Madre Mesenquimatosas , Humanos , Ratones , Animales , Emisiones de Vehículos/toxicidad , Material Particulado/toxicidad , Compuestos Orgánicos , Adipocitos/química , Inflamación , Contaminantes Atmosféricos/toxicidadRESUMEN
Intramuscular fat content is an important determinant of meat quality, and preadipocyte differentiation plays a critical role in intramuscular fat deposition in pigs. However, many types of RNA differentiation, including messenger RNA (mRNA), long non-coding RNA (lncRNA), and circular RNA (circRNA) remain unreported despite their crucial roles in regulating adipogenesis. Chinese Guizhou Congjiang pigs are raised in the Guizhou province of China for their high-quality meat. Therefore, it is important for breeders to explore the mechanisms of proliferation and differentiation of intramuscular adipocytes from the longissimus dorsi muscle of these pigs. In the present study, a transcriptome analysis of intramuscular preadipocytes from Chinese Guizhou Congjiang pigs, including analyses of mRNAs, lncRNAs, and circRNAs at days 0 (D0), 4 (D4), and 8 (D8) was performed. A total of 1,538, 639, and 445 differentially expressed (DE) mRNAs, 479, 192, and 126 DE lncRNAs, and 360, 439, and 304 DE circRNAs were detected between D4 and D0, D8 and D0, and D8 and D4, respectively. Functional analyses identified many significantly enriched RNAs related to lipid deposition, cell differentiation, metabolism processes, and obesity-related diseases, biological processes, and pathways. We identified two lncRNAs (TCONS_00012086 and TCONS_00007245) closely related to fat deposition according to their target genes and tissue expression profiles. Subcellular distribution analysis using quantitative real-time PCR (qRT-PCR) revealed that both TCONS_00012086 and TCONS_00007245 are cytoplasmic lncRNAs. These data provide a genome-wide resource for mRNAs, lncRNAs, and circRNAs potentially involved in Chinese Guizhou Congjiang pig fat metabolism, thus improving our understanding of their function in adipogenesis.
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Adipocitos/citología , MicroARNs/genética , ARN Circular/genética , ARN Largo no Codificante/genética , Análisis de Secuencia de ARN/veterinaria , Adipocitos/química , Adipogénesis , Animales , Diferenciación Celular , Células Cultivadas , China , Grasas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Metabolismo de los Lípidos , Carne/análisis , PorcinosRESUMEN
Risk factors for nonalcoholic hepatic steatosis include obesity and vitamin D deficiency which commonly coexist. Thus, the role of vitamin D signaling in the prevention of hepatic steatosis in the absence of obesity or a "Western" high-fat diet is unclear. These studies were performed to address the role of the adipocyte vitamin D receptor (VDR) in the prevention of hepatic steatosis in mice fed a chow diet containing 5% fat by weight. Female mice with adipocyte VDR ablation (Adipoq-Cre; VDRflox/flox) exhibited a mild increase in weight gain at age 70 days, accompanied by an increase in visceral white adipose tissue (VAT) weight. While they did not exhibit evidence of hepatic inflammation or fibrosis, an increase in hepatic lipid content was observed. This was accompanied by an increase in the hepatic expression of genes involved in fatty acid transport and synthesis, as well as fatty acid oxidation. Markers of hepatic inflammation and fibrosis were unaffected by adipocyte VDR ablation. Consistent with the increase in VAT weight in the Adipoq-Cre; VDRflox/flox mice, higher levels of transcripts encoding adipogenesis-related genes were observed in VAT. In contrast to other models of impaired vitamin D signaling studied in the setting of a high-fat or "Western" diet, the Adipoq-Cre; VDRflox/flox mice do not exhibit hepatic inflammation or fibrosis. These findings suggest that the adipocyte VDR regulates hepatic lipid accumulation, but in the absence of obesity or a high-fat diet, is not required to prevent hepatic inflammation or fibrosis.
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Dieta con Restricción de Grasas , Grasa Intraabdominal/metabolismo , Enfermedad del Hígado Graso no Alcohólico/fisiopatología , Receptores de Calcitriol/fisiología , Adipocitos/química , Animales , Femenino , Metabolismo de los Lípidos/fisiología , Hígado/metabolismo , Ratones , Ratones Noqueados , Enfermedad del Hígado Graso no Alcohólico/prevención & control , Receptores de Calcitriol/deficiencia , Receptores de Calcitriol/genética , Transducción de Señal/fisiología , Vitamina D/metabolismoRESUMEN
Exosomes are secreted into the extracellular space by most cell types and contain various molecular constituents, which play roles in many biological processes. Adipose-derived mesenchymal stem cells (ADSCs) can differentiate into a variety of cell types and secrete a series of paracrine factors through exosomes. ADSC-derived exosomes have shown diagnostic and therapeutic potential in many clinical diseases. The molecular components are critical for their mechanisms. Several methods have been developed for exosome purification, including ultracentrifugation, ultrafiltration, density gradient purification, size-based isolation, polymer precipitation and immuno-affinity purification. Thus, we employed four methods to isolate exosomes from the hADSC culture medium, including ultracentrifugation, size exclusion chromatography, ExoQuick-TC precipitation and ExoQuick-TC ULTRA isolation. Following exosome isolation, we performed quantitative proteomic analysis of the exosome proteins using isobaric tags for relative and absolute quantification (iTRAQ) labelling, combined with 2D-LC-MS/MS. There were 599 universal and 138 stably expressed proteins in hADSC-derived exosomes. We proved that these proteins were potential hADSC-derived exosomes markers, including CD109, CD166, HSPA4, TRAP1, RAB2A, RAB11B and RAB14. From the quantitative proteomic analysis, we demonstrated that hADSC-derived exosome protein expression varied, with lipopolysaccharide (LPS) treatment, in the different isolation methods. Pathway analysis and proliferation, migration and endothelial tube formation assays showed varying effects in cells stimulated with hADSC-derived exosomes from different isolation methods. Our study revealed that different isolation methods might introduce variations in the protein composition in exosomes, which reflects their effects on biological function. The pros and cons of these methods are important points to consider for downstream research applications.
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Fraccionamiento Celular/métodos , Exosomas/química , Células Madre Mesenquimatosas/química , Proteoma/química , Proteómica/métodos , Adipocitos/química , Células Cultivadas , Exosomas/metabolismo , Células Endoteliales de la Vena Umbilical Humana/química , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Células Madre Mesenquimatosas/metabolismo , Redes y Vías MetabólicasRESUMEN
CONTEXT: Healthy hyperplasic (many but smaller fat cells) white adipose tissue (WAT) expansion is mediated by recruitment, proliferation and/or differentiation of new fat cells. This process (adipogenesis) is controlled by transcriptional programs that have been mostly identified in rodents. OBJECTIVE: A systemic investigation of adipogenic human transcription factors (TFs) that are relevant for metabolic conditions has not been revealed previously. METHODS: TFs regulated in WAT by obesity, adipose morphology, cancer cachexia, and insulin resistance were selected from microarrays. Their role in differentiation of human adipose tissue-derived stem cells (hASC) was investigated by RNA interference (RNAi) screen. Lipid accumulation, cell number, and lipolysis were measured for all screened factors (148 TFs). RNA (RNAseq), protein (Western blot) expression, insulin, and catecholamine responsiveness were examined in hASC following siRNA treatment of selected target TFs. RESULTS: Analysis of TFs regulated by metabolic conditions in human WAT revealed that many of them belong to adipogenesis-regulating pathways. The RNAi screen identified 39 genes that affected fat cell differentiation in vitro, where 11 genes were novel. Of the latter JARID2 stood out as being necessary for formation of healthy fat cell metabolic phenotype by regulating expression of multiple fat cell phenotype-specific genes. CONCLUSION: This comprehensive RNAi screening in hASC suggests that a large proportion of WAT TFs that are impacted by metabolic conditions might be important for hyperplastic adipose tissue expansion. The screen also identified JARID2 as a novel TF essential for the development of functional adipocytes.
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Adipocitos/metabolismo , Adipogénesis/genética , Complejo Represivo Polycomb 2/genética , Interferencia de ARN/fisiología , Factores de Transcripción/análisis , Factores de Transcripción/genética , Adipocitos/química , Adipocitos/patología , Tejido Adiposo Blanco/química , Tejido Adiposo Blanco/patología , Adolescente , Secuencia de Bases , Diferenciación Celular/genética , Células Cultivadas , Femenino , Neoplasias Gastrointestinales , Regulación de la Expresión Génica , Humanos , Hiperplasia/genética , Resistencia a la Insulina/genética , Masculino , Obesidad/genética , Complejo Represivo Polycomb 2/fisiología , Células Madre/química , Factores de Transcripción/fisiologíaRESUMEN
Adipose tissue is a primary regulator of energy balance and metabolism. The distribution of adipose tissue depots is of clinical interest because the accumulation of upper-body subcutaneous (ASAT) and visceral adipose tissue (VAT) is associated with cardiometabolic diseases, whereas lower-body glutealfemoral adipose tissue (GFAT) appears to be protective. There is heterogeneity in morphology and metabolism of adipocytes obtained from different regions of the body, but detailed knowledge of the constituent proteins in each depot is lacking. Here, we determined the human adipocyte proteome from ASAT, VAT, and GFAT using high-resolution Sequential Window Acquisition of all Theoretical (SWATH) mass spectrometry proteomics. We quantified 4,220 proteins in adipocytes, and 2,329 proteins were expressed in all three adipose depots. Comparative analysis revealed significant differences between adipocytes from different regions (6% and 8% when comparing VAT vs. ASAT and GFAT, 3% when comparing the subcutaneous adipose tissue depots, ASAT and GFAT), with marked differences in proteins that regulate metabolic functions. The VAT adipocyte proteome was overrepresented with proteins of glycolysis, lipogenesis, oxidative stress, and mitochondrial dysfunction. The GFAT adipocyte proteome predicted the activation of peroxisome proliferator-activated receptor α (PPARα), fatty acid, and branched-chain amino acid (BCAA) oxidation, enhanced tricarboxylic acid (TCA) cycle flux, and oxidative phosphorylation, which was supported by metabolomic data obtained from adipocytes. Together, this proteomic analysis provides an important resource and novel insights that enhance the understanding of metabolic heterogeneity in the regional adipocytes of humans.NEW & NOTEWORTHY Adipocyte metabolism varies depending on anatomical location and the adipocyte protein composition may orchestrate this heterogeneity. We used SWATH proteomics in patient-matched human upper- (visceral and subcutaneous) and lower-body (glutealfemoral) adipocytes and detected 4,220 proteins and distinguishable regional proteomes. Upper-body adipocyte proteins were associated with glycolysis, de novo lipogenesis, mitochondrial dysfunction, and oxidative stress, whereas lower-body adipocyte proteins were associated with enhanced PPARα activation, fatty acid, and BCAA oxidation, TCA cycle flux, and oxidative phosphorylation.
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Adipocitos/metabolismo , Metabolismo Energético/fisiología , Proteoma/análisis , Adipocitos/química , Adipocitos/patología , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Grasa Intraabdominal/metabolismo , Obesidad/metabolismo , Obesidad/patología , Especificidad de Órganos , Proteómica , Grasa Subcutánea/metabolismoRESUMEN
Eurysoloids A (1) and B (2), two novel diastereomeric sesterterpenoids possessing a pentacyclic 5/6/5/10/5 framework with an unusual macrocyclic ether system, were isolated from Eurysolen gracilis Prain. Their structures were unambiguously determined by spectroscopic, single-crystal X-ray diffraction and DP4+ analyses. A plausible biosynthetic pathway for compounds 1 and 2 was proposed. Both compounds exhibited immunosuppressive activity via inhibiting the production of cytokine IFN-γ of T cells, and compound 2 inhibited adipogenesis in 3T3-L1 adipocytes.
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Adipocitos/química , Adipogénesis/efectos de los fármacos , Éter/metabolismo , Lamiaceae/química , Sesterterpenos/farmacología , Células 3T3-L1 , Adipocitos/metabolismo , Animales , Éter/química , Ratones , Estructura Molecular , Sesterterpenos/química , Sesterterpenos/aislamiento & purificaciónRESUMEN
Exosomes are released from a variety of cells to communicate with recipient cells. Exosomes contain microRNAs (miRNAs), which are noncoding RNAs that suppress target genes. Our previous proteomic study (FEBS Open Bio 2016, 6, 816-826) demonstrated that 3T3-L1 adipocytes secrete exosome components as well as growth factors, inspiring us to investigate what type of miRNA is involved in adipocyte-secreted exosomes and what functions they carry out in recipient cells. Here, we profiled miRNAs in 3T3-L1 adipocyte-secreted exosomes and revealed suppression of muscle differentiation by adipocyte-derived exosomes. Through our microarray analysis, we detected over 300 exosomal miRNAs during adipocyte differentiation. Exosomal miRNAs present during adipocyte differentiation included not only pro-adipogenic miRNAs but also miRNAs associated with muscular dystrophy. Gene ontology analysis predicted that the target genes of miRNAs are associated primarily with transcriptional regulation. To further investigate whether adipocyte-secreted exosomes regulate the expression levels of genes involved in muscle differentiation, we treated cultured myoblasts with adipocyte-derived exosome fractions. Intriguingly, the expression levels of myogenic regulatory factors, Myog and Myf6, and other muscle differentiation markers, myosin heavy-chain 3 and insulin-like growth factor 2, were significantly downregulated in myoblasts treated with adipocyte-derived exosomes. Immature adipocyte-derived exosomes exhibited a stronger suppressive effect than mature adipocyte-derived exosomes. Our results suggest that adipocytes suppress the expression levels of muscle differentiation-associated genes in myoblasts via adipocyte-secreted exosomes containing miRNAs.
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Adipocitos/citología , Exosomas/genética , Marcadores Genéticos , MicroARNs/genética , Mioblastos/citología , Células 3T3-L1 , Adipocitos/química , Animales , Regulación hacia Abajo , Perfilación de la Expresión Génica , Factor II del Crecimiento Similar a la Insulina/genética , Ratones , Desarrollo de Músculos , Mioblastos/química , Factores Reguladores Miogénicos/genética , Miogenina/genética , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
Hibernoma is a rare benign adipocytic tumor composed of a proliferation of brown and white fat cells varying in their proportions. The tumor may also contain fat cells resembling lipoblasts, which makes it difficult to distinguish it from atypical lipomatous tumor/well differentiated liposarcoma (ALT/WDLS). Although nuclear expressions of murine double minute 2 (MDM2) and cyclin-dependent kinase 4 (CDK4) are widely used as immunohistochemical surrogate markers for ALT/WDLS, the utility of these proteins in distinguishing between hibernoma and ALT/WDLS still remains to be elucidated. We evaluated immunohistochemical expressions of MDM2 and CDK4 in 10 hibernomas expressing uncoupling protein-1 (UCP-1), a mitochondrial protein transporter consistently expressed in brown fat cells, and lacking MDM2 gene amplification, which was analyzed by fluorescence in situ hybridization (FISH). In contrast to the data previously obtained, nuclear expression of MDM2 was observed in 100% (10/10 cases) of the hibernomas irrespective of the proportion of brown fat cells, whereas no cases were positive for CDK4. The tumors also showed almost concurrent expression of p53 (in 9/10 cases) and ubiquitin-specific-processing protease 7 (USP7) (in 10/10 cases), which deubiquitinates and stabilizes MDM2, potentially resulting in its nuclear expression without MDM2 gene amplification. MDM2 expression may thus be a diagnostic pitfall for hibernoma particularly in differentiating it from ALT/WDLS.
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Adipocitos/química , Biomarcadores de Tumor/análisis , Núcleo Celular/química , Lipoma/química , Proteínas Proto-Oncogénicas c-mdm2/análisis , Adipocitos/patología , Adulto , Anciano , Biomarcadores de Tumor/genética , Núcleo Celular/patología , Quinasa 4 Dependiente de la Ciclina/análisis , Diagnóstico Diferencial , Femenino , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Lipoma/genética , Lipoma/patología , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Proteínas Proto-Oncogénicas c-mdm2/genética , Proteína p53 Supresora de Tumor/análisis , Peptidasa Específica de Ubiquitina 7/análisis , Proteína Desacopladora 1/análisis , Adulto JovenRESUMEN
In recent years, obesity has become a major public health concern. Obesity has been previously associated with lowgrade inflammation and TNFα induction in adipose tissue, which subsequently disrupts adipocyte metabolism. MicroRNAs (miRNAs/miRs) are important metabolic factors and their dysregulation has been associated with obesityrelated metabolic syndromes. In fact, it has been directly suggested that miR424 may be functionally associated with adipogenesis, although its exact role in this process remains unclear. The present study aimed to identify the function of miR424 in adipogenesis. In the present study, miR424 expression levels were analyzed during adipogenesis and the differential expression of this miRNA in the adipose tissue of obese and nonobese children was also assessed. Furthermore, the interaction between miR424 and the adipocytokine TNFα was determined. Finally, miR424 target genes and downstream signaling pathways were predicted via bioinformatics and analyzed by performing a luciferase reporter assay to elucidate the functional mechanisms of miR424 in adipogenesis of visceral adipocytes. The results revealed that the expression levels of miR424 upregulated in the adipose tissue biopsies from obese children compared with the biopsies of nonobese children. However, in cultured adipocytes, the expression levels of miR424 were discovered to be gradually downregulated during the adipogenesis process. TNFα treatment significantly downregulated the expression levels of miR424 via binding to its promoter region and reducing its transcriptional activity. Through bioinformatic prediction analysis, miR424 target genes were analyzed, of which several were identified to be involved in signaling pathways that are known to regulate adipogenesis, such as the Wnt signaling pathway. In conclusion, the present study indicated that miR424 was regulated by TNFα and served an important role in adipogenesis.
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Adipocitos/citología , MicroARNs/genética , Obesidad/genética , Factor de Necrosis Tumoral alfa/metabolismo , Adipocitos/química , Adipogénesis , Estudios de Casos y Controles , Diferenciación Celular , Células Cultivadas , Niño , Humanos , Masculino , Obesidad/metabolismo , Regiones Promotoras Genéticas , Regulación hacia Arriba , Vía de Señalización WntRESUMEN
Determination of the adipogenic potential and behavior of adipose-derived mesenchymal stem/stromal cells (ASCs) is particularly relevant for their potential clinical application in regenerative medicine, especially when regeneration is supported by biomaterials or scaffolds. Scaffolds need to be able to induce tissue repair and limit undesired adipogenic differentiation. Depending on the scaffold employed, determination of cell behavior may be hindered by material interference with staining, which will limit either cells identification or dye quantification. Collagen is a promising biomaterial in regenerative medicine, however, histological analysis of cells cultured on collagen-based scaffolds is challenging. Here we describe a new histological method based on iron hematoxylin combined with Oil red O (ORO) staining, for the determination of the adipogenic differentiation of ASCs cultivated on a collagen-based 2D scaffold. ASCs were seeded on collagen films or plastic, differentiated into adipocytes for 14 days, and then stained with either ORO or iron hematoxylin and ORO combined. The collagen films avidly absorbed the ORO dye; conventional staining and quantification by dye extraction failed to discriminate between differentiated and undifferentiated cells on the films. On the contrary, the iron hematoxylin-ORO combination provided a quantitative and more reliable determination of adipocytes based on single cell count. This method is particularly recommended for determining the adipogenic differentiation potential of ASCs and other cell types grown on highly absorptive materials that need to be validated for their potential use in bioengineering and regenerative medicine.
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Adipocitos/química , Colágeno/química , Células Madre Mesenquimatosas/química , Adipocitos/citología , Compuestos Azo/química , Diferenciación Celular , Células Cultivadas , Hematoxilina/química , Humanos , Hierro/química , Células Madre Mesenquimatosas/citología , Coloración y EtiquetadoRESUMEN
The cellular response to fluctuations in blood glucose levels consists of integrative regulation of cell glucose uptake and glucose utilization in the cytosol, resulting in altered levels of glucose in the cytosol. Cytosolic glucose is difficult to be measured in the intact tissue, however recently methods have become available that allow measurements of glucose in single living cells with fluorescence resonance energy transfer (FRET) based protein sensors. By studying the dynamics of cytosolic glucose levels in different experimental settings, we can gain insights into the properties of plasma membrane permeability to glucose and glucose utilization in the cytosol, and how these processes are modulated by different environmental conditions, agents and enzymes. In this review, we compare the cytosolic regulation of glucose in adipocytes and astrocytes - two important regulators of energy balance and glucose homeostasis in whole body and brain, respectively, with particular emphasis on the data obtained with FRET based protein sensors as well as other biochemical and molecular approaches.
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Adipocitos/química , Astrocitos/química , Citosol/química , Transferencia Resonante de Energía de Fluorescencia , Glucosa/análisis , Nanopartículas/química , Adipocitos/metabolismo , Astrocitos/metabolismo , Células Cultivadas , Citosol/metabolismo , Glucosa/metabolismo , HumanosRESUMEN
This study investigated the effect of ELOVL6 (elongation of very long chain fatty acids protein 6) and its underlying mechanism on lipid metabolism in bovine adipocytes. The ELOVL6 gene was overexpressed in bovine adipocytes by adenoviruses, and RNA sequencing was performed. Overexpression of ELOVL6 showed reduced proportions of C14:0 (Myristic) and C16:0 (palmitate) fatty acids and increased proportions of C18.0 (stearate) and C20:4n6 (arachidonic) fatty acids in adipocytes. In addition, a total of 2170 differentially expressed genes (DEGs) were found, containing 1802 up-regulated and 368 down-regulated genes. KEGG pathway analysis revealed that the down-regulated genes were linked with the regulation of lipolysis and the Wnt signaling pathway. The up-regulated genes were mainly involved in the FoxO signaling pathway; the PI3K-Akt signaling pathway; and the cAMP signaling pathway. In conclusion, our results suggest that ELOVL6 could affect the fatty acid composition in bovine adipocytes. We identified numerous related DEGs and pathways, which may provide a basis for studying the function and molecular mechanism of the ELOVL6 gene in regulating lipid metabolism.
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Adipocitos/metabolismo , Bovinos/metabolismo , Elongasas de Ácidos Grasos/metabolismo , Metabolismo de los Lípidos , Adipocitos/química , Animales , Bovinos/genética , Células Cultivadas , Elongasas de Ácidos Grasos/química , Elongasas de Ácidos Grasos/genética , Ácidos Grasos/análisis , Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Metabolismo de los Lípidos/genética , Lipólisis/genética , Filogenia , Alineación de Secuencia , Análisis de Secuencia de ProteínaRESUMEN
Populations of industrialized countries have registered a dramatically increasing prevalence in obesity for many years. Despite continuous research, mechanisms involved in the storage and utilization of chemical energy in adipocytes are still under investigation. Adipocytes have the task to store excessive energy in the form of triacylglycerols (TG) and it is already well-known that the fatty acyl composition of TG is largely determined by the composition of the diet. In contrast to TG, the composition of adipocyte phospholipids was less comprehensively investigated. In this study, the compositions of the most abundant phospholipid classes of 3T3-L1 undifferentiated (preadipocytes) and differentiated cells (adipocytes) were determined. The lipid fractions were isolated by normal phase high-performance thin-layer chromatography and subsequently analyzed by electrospray ionization mass spectrometry. Additionally, the fatty acyl (FA) compositions were determined by gas chromatography. The positions of the FA residues were additionally confirmed by phospholipase A2 digestion. The advantages and disadvantages of the different analytical approaches will be discussed. It will be shown that undifferentiated 3T3-L1 and mature adipocytes differ extremely regarding their compositions. This goes along with an increase in odd-chain fatty acids. Graphical abstract.
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Adipocitos/química , Adipocitos/citología , Metabolismo de los Lípidos , Lípidos/química , Células 3T3-L1 , Adipocitos/metabolismo , Animales , Diferenciación Celular , Cromatografía en Capa Delgada/métodos , Ácidos Grasos/química , Ácidos Grasos/metabolismo , Cromatografía de Gases y Espectrometría de Masas/métodos , Ratones , Triglicéridos/química , Triglicéridos/metabolismoRESUMEN
BACKGROUND AND OBJECTIVE: Obesity is highly complicated by hypertension and hyperglycemia. In particular, it has been proposed that obesity-related hypertension is caused by adipocyte-derived factors that are recognized as undetermined proteins secreted from adipocytes. Adipocyte-derived factors have been known to be related to aldosterone secretion in the adrenal gland. So far, Wnt proteins, CTRP-1, VLDL, LDL, HDL and leptin have been demonstrated to stimulate aldosterone secretion. In contrast, it has not yet been clarified whether adipocyte-derived factors also affect adrenal cortisol secretion. METHODS AND RESULTS: In the present study, we investigated the effect of adipocyte-derived factors on cortisol synthase gene CYP11B1 mRNA expression in vitro study using adrenocortical carcinoma H295R cells and mouse fibroblast 3T3-L1cells. Interestingly, adipocyte-derived factors were demonstrated to have the ability to stimulate CYP11B1 mRNA expression. CONCLUSION: Since CYP11B1 is well known as a limiting enzyme of cortisol synthesis, our study suggests that adipocyte-derived factors may stimulate cortisol secretion, as well as aldosterone secretion. Taken together, adipocyte-derived factors may be the cause of metabolic syndrome due to their stimulating effects on aldosterone/cortisol secretion. Therefore, the innovation of novel drugs against them may possibly be a new approach against metabolic syndrome.
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Adipocitos/química , Corteza Suprarrenal/efectos de los fármacos , Citocromo P-450 CYP11B2/biosíntesis , Esteroide 11-beta-Hidroxilasa/biosíntesis , Corteza Suprarrenal/metabolismo , Neoplasias de la Corteza Suprarrenal/metabolismo , Neoplasias de la Corteza Suprarrenal/patología , Animales , Carcinoma/metabolismo , Carcinoma/patología , Línea Celular , Línea Celular Tumoral , Citocromo P-450 CYP11B2/genética , Fibroblastos , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Hidrocortisona/metabolismo , Leptina/metabolismo , Leptina/farmacología , Lipoproteínas LDL/metabolismo , Lipoproteínas LDL/farmacología , Ratones , Proteínas/genética , Proteínas/fisiología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Esteroide 11-beta-Hidroxilasa/genética , Proteínas Wnt/metabolismo , Proteínas Wnt/farmacología , Zona Fascicular/efectos de los fármacos , Zona Fascicular/metabolismoRESUMEN
The ability to discriminately analyze the chemical constituents of single cells and organelles is highly sought after and necessary to establish true biomarkers. Some major challenges of individual cell analysis include requirement and expenditure of a large sample of cells as well as extensive extraction and separation techniques. Here, we describe methods to perform individual cell and organelle extractions of both tissues and cells in vitro using nanomanipulation coupled to mass spectrometry. Lipid profiles display heterogeneity from extracted adipocytes and lipid droplets, demonstrating the necessity for single cell analysis. The application of these techniques can be applied to other cell and organelle types for selective and thorough monitoring of disease progression and biomarker discovery.
Asunto(s)
Lipidómica/métodos , Lípidos/análisis , Micromanipulación/métodos , Análisis de la Célula Individual/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Células 3T3-L1 , Adipocitos/química , Adipocitos/citología , Adipocitos/metabolismo , Adipogénesis , Animales , Células Cultivadas , Diseño de Equipo , Humanos , Gotas Lipídicas/química , Gotas Lipídicas/metabolismo , Metabolismo de los Lípidos , Lipidómica/instrumentación , Ratones , Micromanipulación/instrumentación , Análisis de la Célula Individual/instrumentación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/instrumentaciónRESUMEN
BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent stem cells that are able to differentiate into several cell types, including cartilage, fat, and bone. As a common progenitor, MSC differentiation has to be tightly regulated to maintain the balance of their differentiation commitment. It has been reported that the decision process of MSCs into fat and bone cells is competing and reciprocal. Several factors have been suggested as critical factors that affect adipo-osteogenic decision, including melatonin and smad4. Yes-associated protein (YAP) is an important effector protein in the Hippo signaling pathway that acts as a transcriptional regulator by activating the transcription of the genes involved in cell proliferation and anti-apoptosis. The non-canonical role of YAP in regulating bone homeostasis by promoting osteogenesis and suppressing adipogenesis was recently demonstrated in a mouse model. However, it is unclear whether YAP is also crucial for modulating human MSC differentiation to fat and bone. METHODS: The expression level of YAP during MSC differentiation was modulated using pharmaceutical molecule and genetic experiments through gain- and loss-of-function approaches. RESULTS: We demonstrated for the first time that YAP has a non-canonical role in regulating the balance of adipo-osteogenic differentiation of human MSCs. The result from synchrotron radiation-based Fourier transform infrared (FTIR) microspectroscopy showed unique metabolic fingerprints generated from YAP-targeted differentiated cells that were clearly distinguished from non-manipulated control. CONCLUSIONS: These results, thus, identify YAP as an important effector protein that regulates human MSC differentiation to fat and bone and suggests the use of FTIR microspectroscopy as a promising technique in stem cell research.
Asunto(s)
Adipogénesis , Proteínas de Ciclo Celular/metabolismo , Diferenciación Celular , Osteogénesis , Factores de Transcripción/metabolismo , Adipocitos/química , Adipocitos/citología , Adipocitos/metabolismo , Adipogénesis/efectos de los fármacos , Proteínas de Ciclo Celular/genética , Diferenciación Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Dobutamina/farmacología , Humanos , Inmunofenotipificación , Lisofosfolípidos/farmacología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/química , Osteoblastos/citología , Osteoblastos/metabolismo , Osteogénesis/efectos de los fármacos , Análisis de Componente Principal , Espectroscopía Infrarroja por Transformada de Fourier , Factores de Transcripción/genética , Cordón Umbilical/citologíaRESUMEN
The 3T3-L1 pre-adipocyte cell line is widely used to study the fat cell differentiation in vitro. Researchers also use this cell model to study obesity and insulin resistance. We surveyed the literature, the gene expression omnibus and the sequence read archive for RNA-Seq and ChIP-Seq datasets of MDI-induced 3T3-L1 differentiating cells sampled at one or more time points. The metadata of the relevant datasets were manually curated using unified language across the original studies. The raw reads were collected and pre-processed using a reproducible state-of-the-art pipeline. The final datasets are presented as reads count in genes for the RNA-Seq and reads count in peaks for the ChIP-Seq dataset. The curated datasets are available as two Bioconductor experimental data packages curatedAdipoRNA and curatedAdipoChIP. In addition, the packages document the source code of the data collection and the pre-processing pipelines. Here, we provide a descriptive analysis of the datasets with context and technical validation.
Asunto(s)
Adipocitos/citología , Proteínas de Unión al ADN/genética , Perfilación de la Expresión Génica/métodos , 1-Metil-3-Isobutilxantina/farmacología , Células 3T3-L1 , Adipocitos/química , Adipocitos/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Secuenciación de Inmunoprecipitación de Cromatina , Dexametasona/farmacología , Secuenciación de Nucleótidos de Alto Rendimiento , Insulina/farmacología , Ratones , Modelos Moleculares , Análisis de Secuencia de ARNRESUMEN
OBJECTIVE: The aim of this study was to determine the potential association of foot pain and plasmatic adipocytes as physiological biomarkers of childhood obesity with the incidence of flatfoot in a cohort of Egyptian school children aged 6 -12 years. METHODS: A total of 550 Egyptian schoolchildren (220 boys and 330 girls) aged 6-12 years were randomly invited to participate in this descriptive survey analysis. For all children, we assessed the diagnosis and severity of flatfoot as well as plasma adipocytes, as well as adiponectin, leptin, resistin, IL-6, and TNF-α, using the Dennis method and immunoassay techniques respectively. Foot pain was assessed by using a standard VAS of 100 mm and Faces Pain Scale, respectively. RESULTS: Flat foot was predicted in 30.4% of school-age children, most of them showed a higher frequency of overweight (33.3%) and obesity (62.5%). Boys showed higher ranges of flat foot than girls. Foot pain significantly correlated with flat foot and obesity among the studied populations. In overweight-obese children, plasmatic adipocyte variables, as well as adiponectin, leptin, resistin, IL-6, TNF-α showed significant correlations with foot stance, especially in boys. Also, the studied adipocyte variables along with BMI, age, gender explained about~65% of the variance of flatfoot with pain among our school-age students. CONCLUSION: Foot pain showed an association with flat foot and childhood obesity in 30.4% of school-age students (6-12 years). Foot pain was shown to correlate positively with the incidence of flat foot and changes in adiposity markers, as well as adiponectin, leptin, resistin, Il-6, TNF-α.