CONTEXT: Adipose tissue distribution is a key factor influencing metabolic health and risk in obesity-associated comorbidities. OBJECTIVE: Here we aim to compare the proteomic profiles of mature adipocytes from different depots. METHODS: Abdominal subcutaneous (SA) and omental visceral adipocytes (VA) were isolated from paired adipose tissue biopsies obtained during bariatric surgery on 19 severely obese women (body mass index > 30 kg/m2) and analyzed using state-of-the-art mass spectrometry. Differential expression analysis and weighted gene co-expression network analysis (WGCNA) were performed to investigate proteome signature properties and to examine a possible association of the protein expression with the clinical data. RESULTS: We identified 3686 protein groups and found 1140 differentially expressed proteins (adj. P value < 0.05), of which 576 proteins were upregulated in SA and 564 in VA samples. We provide a global protein profile of abdominal SA and omental VA, present the most differentially expressed pathways and processes distinguishing SA from VA, and correlate them with clinical and body composition data. We show that SA are significantly more active in processes linked to vesicular transport and secretion, and to increased lipid metabolism activity. Conversely, the expression of proteins involved in the mitochondrial energy metabolism and translational or biosynthetic activity is higher in VA. CONCLUSION: Our analysis represents a valuable resource of protein expression profiles in abdominal SA and omental VA, highlighting key differences in their role in obesity.
- MeSH
- bariatrická chirurgie MeSH
- dospělí MeSH
- genové regulační sítě MeSH
- lidé středního věku MeSH
- lidé MeSH
- morbidní obezita metabolismus patologie chirurgie MeSH
- nitrobřišní tuk cytologie metabolismus patologie MeSH
- omentum cytologie metabolismus patologie chirurgie MeSH
- podkožní břišní tuk cytologie metabolismus patologie MeSH
- proteomika MeSH
- tukové buňky metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
- práce podpořená grantem MeSH
Aim of the study: The aim was to perform a morphometric analysis of subcutaneous and visceral adipose tissue of morbidly obese women and to determine the relationship between adipocyte size and the development of type 2 diabetes (T2D). Materials and methods: White adipose tissue of morbidly obese women was obtained from subcutaneous and omental adipose tissue during bariatric surgery. The same tissues were obtained at judicial autopsy in non-obese (lean) non-diabetic patients. The harvested tissue was embedded in paraffin and 5 μm thick hematoxylin-eosin stained sections were analyzed by the Olympus cellSens system. Statistical evaluation was performed by GraphPad Prism 6.1 software. Results: We found a relationship between adipocyte size and the presence of T2D. The most pronounced changes were seen in visceral adipocytes (cell diameter increased from 61.9 μm in controls to 79.5 μm in patients with T2D). Also, the size of the subcutaneous adipocytes increased against the control. A statistically significant difference between diabetic and non-diabetic patients was not proven in subcutaneous adipocytes. We also observed differences in the distribution of adipocyte mean diameters. Whilst in the control group there was a normal (Gaussian) distribution, in the morbidly obese we found an asymmetric distribution with a positive skewness to the right. Conclusion: We have demonstrated that in morbidly obese women a significant increase in visceral adipocyte size is associated with the development of both insulin resistance and T2D.
- MeSH
- bílé tukové buňky cytologie MeSH
- diabetes mellitus 2. typu patofyziologie MeSH
- dospělí MeSH
- lidé MeSH
- morbidní obezita * chirurgie patofyziologie MeSH
- nitrobřišní tuk cytologie MeSH
- podkožní břišní tuk cytologie patofyziologie MeSH
- tukové buňky cytologie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
Data from experimental animal models and in vitro studies suggest that both hyperlipoproteinemia and obesity predispose to development of proinflammatory pathways of macrophages within adipose tissue. The aim of this study was to analyze whether non-HDL cholesterol concentration in healthy living kidney donors (LKDs) is related to the number and phenotype of proinflammatory macrophages in visceral and subcutaneous adipose tissue. Adipose tissue samples were collected by cleansing the kidney grafts of LKDs obtained peroperatively. The stromal vascular fractions of these tissues were analyzed by flow cytometry. Proinflammatory macrophages were defined as CD14(+) cells coexpressing CD16(+) and high-expression CD36 as well (CD14(+)CD16(+)CD36(+++)), while CD16 negativity and CD163 positivity identified alternatively stimulated, anti-inflammatory macrophages. Non-HDL cholesterol concentration positively correlated to proinflammatory macrophages within visceral adipose tissue, with increased strength with more precise phenotype determination. On the contrary, the proportion of alternatively stimulated macrophages correlated negatively with non-HDL cholesterol. The present study suggests a relationship of non-HDL cholesterol concentration to the number and phenotype proportion of macrophages in visceral adipose tissue of healthy humans.
- MeSH
- CD antigeny metabolismus MeSH
- cholesterol metabolismus MeSH
- dospělí MeSH
- ledviny MeSH
- lidé středního věku MeSH
- lidé MeSH
- makrofágy cytologie metabolismus MeSH
- nitrobřišní tuk cytologie metabolismus MeSH
- transplantace ledvin MeSH
- žijící dárci * MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND AND AIMS: Macrophages play important roles in adipose tissue inflammation and its consequences. Unfortunately, a detailed description of the macrophage phenotypes in different human adipose tissues is not available. SUBJECTS AND METHODS: Subcutaneous, visceral and perivascular adipose tissues were obtained from 52 living kidney donors during live donor nephrectomy. Stromal vascular fractions were isolated, and the macrophage phenotypes were analyzed by flow cytometry using surface markers (CD14, CD16, CD36, and CD163). RESULTS: In addition to CD16 positivity, pro-inflammatory macrophages also display high scavenger receptor CD36 expression. The great majority of CD16 negative macrophages express the anti-inflammatory CD163 marker. The presence of pro-inflammatory macrophages was almost twice as high in visceral (p < 0.0001) and perivascular (p < 0.0001) adipose tissues than in subcutaneous tissue. This difference was substantially more pronounced in the postmenopausal women subgroup, consequentlly, the total difference was driven by this subgroup. CONCLUSION: We obtained detailed information about M1 and M2 macrophage phenotypes in human adipose tissue. The visceral and perivascular adipose tissues had substantially higher pro-inflammatory characteristics than the subcutaneous tissue. The higher proportion of pro-inflammatory macrophages in the visceral adipose tissue of postmenopausal women might be related to an increased cardiovascular risk.
- MeSH
- dárci tkání MeSH
- fenotyp MeSH
- lidé středního věku MeSH
- lidé MeSH
- makrofágy cytologie MeSH
- nitrobřišní tuk cytologie MeSH
- podkožní tuk cytologie MeSH
- průtoková cytometrie MeSH
- separace buněk MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
BACKGROUND: Our study focused on the ability of epicardial adipocytes to produce bioactive substances and compare the extent of this production with the production of adipokines in visceral adipocytes, which are well known endocrine cells capable of contributing to the development of atherosclerosis. MATERIAL AND METHODS: The gene expression of human cytokines (IL-6, IL-8, IL-18, RANTES and MCP-1) and adipokines (leptin and adiponectin) was measured in primary cell lines of epicardial and visceral adipocytes, both in undifferentiated and mature statuses, after a 21-day-long differentiation protocol. Each condition was assayed in triplicate in two independent primary cell lines obtained from two different donors. RESULTS: The epicardial preadipocytes showed an increased expression of IL-8 (3.25-fold, p<0.05) compared with visceral preadipocytes. The expression of the atheroprotective adiponectin in epicardial preadipocytes was minimal compared with the expression in visceral preadipocytes (p<0.0001). Moreover, the expression of the genes of interest was dependent on the differentiation degree and cell origin. We observed an altered expression of the proinflammatory genes IL-8 (0.016-fold, p<0.01) and MCP-1 (0.19-fold, p<0.05) in differentiated epicardial adipocytes compared with undifferentiated adipocytes. The epicardial adipocytes showed an increased expression of IL-6 (8.13-fold, p<0.05) compared with the visceral adipocytes. CONCLUSION: Our results suggest that epicardial adipocytes substantially differ from visceral adipocytes and might locally contribute to the pathogenesis of coronary atherosclerosis.
- MeSH
- buněčná diferenciace fyziologie MeSH
- chemokin CCL2 genetika MeSH
- chemokin CCL5 genetika MeSH
- cytokiny genetika MeSH
- dospělí MeSH
- exprese genu fyziologie MeSH
- interleukin-18 genetika MeSH
- interleukin-6 genetika MeSH
- interleukin-8 genetika MeSH
- kultivované buňky MeSH
- lidé MeSH
- nitrobřišní tuk cytologie fyziologie MeSH
- perikard cytologie fyziologie MeSH
- pilotní projekty MeSH
- tukové buňky cytologie fyziologie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
AIMS/HYPOTHESIS: Our goal was to identify a set of human adipose tissue macrophage (ATM)-specific markers and investigate whether their gene expression in subcutaneous adipose tissue (SAT) as well as in visceral adipose tissue (VAT) is related to obesity and to the occurrence of the metabolic syndrome. METHODS: ATM-specific markers were identified by DNA microarray analysis of adipose tissue cell types isolated from SAT of lean and obese individuals. We then analysed gene expression of these markers by reverse transcription quantitative PCR in paired samples of SAT and VAT from 53 women stratified into four groups (lean, overweight, obese and obese with the metabolic syndrome). Anthropometric measurements, euglycaemic-hyperinsulinaemic clamp, blood analysis and computed tomography scans were performed. RESULTS: A panel of 24 genes was selected as ATM-specific markers based on overexpression in ATM compared with other adipose tissue cell types. In SAT and VAT, gene expression of ATM markers was lowest in lean and highest in the metabolic syndrome group. mRNA levels in the two fat depots were negatively correlated with glucose disposal rate and positively associated with indices of adiposity and the metabolic syndrome. CONCLUSIONS/INTERPRETATION: In humans, expression of ATM-specific genes increases with the degree of adiposity and correlates with markers of insulin resistance and the metabolic syndrome to a similar degree in SAT and in VAT.
- MeSH
- dospělí MeSH
- kultivované buňky MeSH
- lidé středního věku MeSH
- lidé MeSH
- makrofágy metabolismus MeSH
- metabolický syndrom metabolismus MeSH
- mladý dospělý MeSH
- nadváha metabolismus MeSH
- nitrobřišní tuk cytologie metabolismus MeSH
- obezita metabolismus MeSH
- podkožní tuk cytologie metabolismus MeSH
- senioři MeSH
- tuková tkáň cytologie metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- abdominální obezita metabolismus MeSH
- ateroskleróza metabolismus MeSH
- diabetes mellitus 2. typu metabolismus MeSH
- energetický metabolismus fyziologie genetika MeSH
- genetická predispozice k nemoci genetika MeSH
- inzulinová rezistence fyziologie MeSH
- lidé MeSH
- metabolický syndrom metabolismus MeSH
- nitrobřišní tuk cytologie imunologie metabolismus MeSH
- obezita diagnóza klasifikace metabolismus MeSH
- oxidační stres imunologie MeSH
- střevní sliznice chemie mikrobiologie MeSH
- xenobiotika metabolismus škodlivé účinky MeSH
- Check Tag
- lidé MeSH
