Abstract:In numerous body locations, muscle and adipose tissue are in close contact. Both tissues are endocrine organs that release cytokines, playing a crutial role in the control of tissue homeostasis in health and diseases. Within this context, the identification of the signals involved in muscle–fat crosstalk has been a hot topic over the last 15 years. Recently, it has been discovered that adipose tissue and muscles can release information embedded in lipid-derived nanovesicles called ‘extracellular vesicles’ (EVs… Show more
“…However, few studies on the exosomes in meat products have been reported. Exosomes are present in muscle and fat of animals in vivo is an indisputable fact [20]. The process of exosomegenesis involves the invaginations of the plasma membrane and the formation of intracellular multivesicular bodies [3].…”
In this study, we extracted exosomes from cooked meat by ultra-high-speed centrifugation. Exosome had an average of size of 70.29 nm. Flow cytometry demonstrated that the positive rate of exosomal surface marker CD63 and CD81 were 84.5% and 95.9%. microRNA sequencing revealed the exosomal microRNA were differences among porcine muscle, fat and liver. The mice plasma levels of miR-1, miR-133a-3p, miR-206 and miR-99a were increase with varying degrees after drinking water with exosomes. GTT and ITT suggest that abnormal glucose metabolism and insulin resistance in mice. Moreover, the lipid droplets were significant increased in the liver were also observed. Transcriptome analysis identified 446 differentially expressed genes in liver. Functional enrichment analysis found that metabolic pathway were most significantly enriched. microRNA may function as a critical regulator involved in the metabolic disorder of mice. This study suggests that the exosomal microRNAs from meat products has the potential to adversely affect health.
“…However, few studies on the exosomes in meat products have been reported. Exosomes are present in muscle and fat of animals in vivo is an indisputable fact [20]. The process of exosomegenesis involves the invaginations of the plasma membrane and the formation of intracellular multivesicular bodies [3].…”
In this study, we extracted exosomes from cooked meat by ultra-high-speed centrifugation. Exosome had an average of size of 70.29 nm. Flow cytometry demonstrated that the positive rate of exosomal surface marker CD63 and CD81 were 84.5% and 95.9%. microRNA sequencing revealed the exosomal microRNA were differences among porcine muscle, fat and liver. The mice plasma levels of miR-1, miR-133a-3p, miR-206 and miR-99a were increase with varying degrees after drinking water with exosomes. GTT and ITT suggest that abnormal glucose metabolism and insulin resistance in mice. Moreover, the lipid droplets were significant increased in the liver were also observed. Transcriptome analysis identified 446 differentially expressed genes in liver. Functional enrichment analysis found that metabolic pathway were most significantly enriched. microRNA may function as a critical regulator involved in the metabolic disorder of mice. This study suggests that the exosomal microRNAs from meat products has the potential to adversely affect health.
“…In 2009, adipose-derived exosomes were first found, which can act as a mode of communication between AT and macrophages [ 53 ]. After that, accumulating evidence indicated that adipose-derived exosomes can transfer specific cargoes from cell to cell via paracrine or endocrine signaling, thereby regulating metabolic inflammation, energy metabolism, and insulin sensitivity of skeletal muscle [ 24 , 54 ]. At present, the study of AT-derived exosomes on skeletal muscle mainly focuses on the research of the cargoes (proteins and miRNAs) contained in the exosomes.…”
Section: Adipose-derived Exosomes In Adipocyte–myocyte Crosstalkmentioning
confidence: 99%
“…In the pathological context, insulin resistance is a key component in the etiology of type 2 diabetes. As the major insulin-sensitive tissues, skeletal muscle was proved to be regulated by adipose-derived exosomes with their cargo during the development of insulin resistance [ 24 ]. Proteomic characterization of adipocytic exosomes from diabetes animal models (OLETF) and controls (LETO) were firstly performed in rats.…”
Section: Proteinsmentioning
confidence: 99%
“…Notably, exosome-carried contents such as nucleic acids, proteins, and lipids are highly heterogenous, depending on the origin and physiopathologic conditions of donor cells [ 23 ]. Considering the extremely secretory activity of AT, there is growing interest in studying the characteristic and function of adipose-secreted exosomes, as well as their roles in adipocyte-muscle interaction [ 24 ]. In this review, we introduce the origin and trafficking of adipose-secreted exosomes and their recent advances in research on skeletal muscle pathophysiology.…”
Due to its prominent secretory activity, adipose tissue (AT) is now considered a major player in the crosstalk between organs, especially with skeletal muscle. In which, exosomes are effective carriers for the intercellular material transfer of a wide range of molecules that can influence a series of physiological and pathological processes in recipient cells. Considering their underlying roles, the regulatory mechanisms of adipose-secreted exosomes and their cellular crosstalk with skeletal muscle have received great attention in the field. In this review, we describe what is currently known of adipose-secreted exosomes, as well as their applications in skeletal muscle pathophysiology.
“…Despite evidence suggesting a diverse range of distinct regulatory functions for myoblast and myotube SM-EVs, the majority of studies to date have typically isolated heterogeneous EV fractions using protocols such as ultracentrifugation or precipitation that have often not accounted for the presence and contribution of co-isolated components such as lipoproteins and RNA binding proteins (e.g. AGO2) routinely identi ed in EV studies due to the lack of an optimal isolation method (23)(24)(25). As such, optimising effective SM-EV isolation methods with high levels of purity will be important if we are to accurately de ne their composition and functional effects in physiological and pathophysiological systems.…”
Background: Skeletal muscle extracellular vesicles (SM-EVs) regulate gene expression events in myogenic differentiation. Optimising effective SM-EV isolation methods offering high levels of purity will be important to accurately define their composition and functionality. Size-exclusion chromatography (SEC) applied in combination with ultrafiltration (UF) has the potential to increase sample throughput, scalability and selectivity. However, an optimal UF+SEC methodology has not been tested for the isolation of myotube derived EVs. Our aim was to compare two different UF protocols and define an optimal window of SEC fractions to maximise SM-EVs recovery and sample purity.
Methods: C2C12 myotube conditioned medium was pre-concentrated using Amicon® Ultra 15 or Vivaspin®20, 100KDa UF columns and processed by SEC (IZON, qEV 70nm). The resulting thirty fractions obtained were individually analysed to identify an optimal fraction window for EV recovery.
Results: EV markers Alix and TSG101 could be detected up to fraction 13, while CD9 and Annexin A2 only up to fraction 6. ApoA1+ lipoprotein contaminants were detected from fraction 6 onwards for both protocols. Amicon and Vivaspin UF preconcentration protocols led to qualitative and quantitative variations in EV marker profiles and purity. Eliminating lipoprotein co-isolation by reducing the SEC fraction window resulted in a net loss of particles, but increased measures of sample purity and had only a negligible impact on the presence of EV marker proteins.
Conclusion: In conclusion, this study developed optimal UF+SEC protocols for the isolation of SM-EVs based on sample purity (fractions 1-5) and total abundance (fractions 2-10). The resulting protocols will be valuable in isolating highly pure SM-EV preparations for biomarker studies.
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