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Mus musculus

ABSTRACT: miRNA profiles within lean and obese adipose tissue macrophages-derived exosomes

PROVIDER: PRJNA382562 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5443730.fastq.gz	Fastqsanger.gz
	SRR5443721.fastq.gz	Fastqsanger.gz
	SRR5443731.fastq.gz	Fastqsanger.gz
	SRR5443722.fastq.gz	Fastqsanger.gz
	SRR5443732.fastq.gz	Fastqsanger.gz

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miRNA profiles within lean and obese adipose tissue macrophages-derived exosomes

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Project description:Affymetrix miRNA 3.0 array profiling of adipocyte-derived exosomes from obese and lean human subjects. We used miRNA arrays to profile exosomes shed from obese and lean human subcutaneous fat that was cultured for 60 minutes. Human obese and lean subcutaneous fat were surgically acquired, dissected, and promptly cultured for 60 minutes. We used the culture supernatants for exosome purification and isolation using ExoQuick-TC Precipitation Solution.

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Project description:Affymetrix miRNA 3.0 array profiling of adipocyte-derived exosomes from obese and lean human subjects We used miRNA arrays to profile exosomes shed from obese and lean human visceral and subcutaneous fat that was cultured for 60 minutes. Human obese and lean visceral and subcutaneous fat were surgically acquired, dissected, and promptly cultured for 60 minutes. We used the culture supernatants for exosome purification and isolation using ExoQuick-TC Precipitation Solution

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Project description:In mammals, expansion of adipose tissue mass induces accumulation of adipose tissue macrophages (ATMs). We isolated CD11c- (FB) and CD11c+ (FBC) perigonadal ATMs from SVCs of lean (C57BL/6J Lep +/+) and obese leptin-deficient (C57BL/6J Lep ob/ob) mice. We used expression microarrays to generate transcription profiles of perigonadal ATMs from lean (C57BL/6J Lep +/+) and obese (C57BL/6J Lep ob/ob) mice. Profiling purified FBs and FBCs, we identified 521 transcripts whose expression was differentially (nominal p-value < 0.01) expressed between FBs from lean and obese mice and 1509 genes whose expression was differentially (nominal p-value <0.01) expressed between FBC from lean and obese mice RNA was isolated from sorted FBC (F4/80+, CD11b+, CD11c+) cells and FB ( F4/80+, CD11b+, CD11c-) cells and using RNeasy micro-kits (Qiagen), using a PicoPure RNA isolation kit then amplified two-rounds. Labeled cRNA Mouse Genome 430 2.0 arrays (purified FB and FBC adipose tissue macrophages. There was a total of sixteen samples. FB and FBC populations were isolated from 4 lean and 4 obese mice.

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Adipose tissue stromal cells from lean, obese, and formerly obese mice

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Project description:Background and Aims: Insulin resistance is a key factor in the pathogenesis of NAFLD. We evaluated the importance of subcutaneous abdominal adipose tissue (SAAT) inflammation and both plasma and SAAT-derived exosomes in regulating insulin sensitivity in people with obesity and NAFLD. Methods: Adipose tissue inflammation (macrophage and T cell content and gene expression of proinflammatory cytokines), liver and whole-body insulin sensitivity (assessed by a hyperinsulinemic-euglycemic clamp and glucose tracer infusion), and 24-hour serial plasma cytokine concentrations were evaluated in three groups stratified by adiposity, insulin sensitivity and intrahepatic triglyceride (IHTG) content: 1) metabolically-healthy lean (LEAN; n=14); 2) metabolically-healthy obese with normal IHTG content (OB-NL; n=28); and 3) metabolically-unhealthy obese with NAFLD (OB-NAFLD; n=28). The effect of plasma and SAAT-derived exosomes on insulin action (insulin-stimulated Akt phosphorylation) in human skeletal muscle myotubes was assessed in a subset of participants. Results: Proinflammatory macrophages, proinflammatory CD4 and CD8 T cell populations, and gene expression of several cytokines in SAAT were greater in the OB-NAFLD than the OB-NL and LEAN groups. However, with the exception of PAI-1, which was greater in the OB-NAFLD than the LEAN and OB-NL groups, 24-hour plasma cytokine concentration areas-under-the-curve (AUC) were not different between groups. The percentage of proinflammatory macrophages and plasma PAI-1 concentration AUC were inversely correlated with both hepatic and whole-body insulin sensitivity. Compared with exosomes from OB-NL participants, plasma and SAAT-derived exosomes obtained from the OB-NAFLD group impaired insulin action in myotubes. Conclusion: These results suggest SAAT-derived exosomes and PAI-1 are involved in the pathogenesis of systemic insulin resistance in people with obesity and NAFLD. ClinicalTrials.gov number: NCT02706262.

2021-10-01 | GSE156906 | GEO

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