The selection of proper reference genes and materials is critical in the design of PCR experiments, especially for differential expression studies. In this study, we propose a method to identify robust endogenous control miRNAs in the visceral adipose tissue of C57BL/6J mice with non-alcoholic fatty liver disease induced by alternating Western and control diets. This study outlines a comprehensive methodology for the analysis of microRNA endogenous controls using microfluidic cards in conjunction with miRNA profiling through small RNA sequencing and subsequent validation by quantitative PCR and the RefFinder algorithm. Criteria included were fold change, p-value, reads per million, and gene stability assessment. A set of six putative endogenous microRNAs was identified (miR-331-3p, let-7a-5p, miR-1839-5p, miR-151a-5p, let-7d-5p, and let-7c-5p). Subsequent validation and analysis using the RefFinder algorithm assessed the stability of the selected genes, and a combination of the three most stable endogenous miRNA controls (miR-331-3p, let-7a- 5p, and miR-1839-5p) exhibiting consistent expression patterns with minimal variability was set. Given the absence of universal endogenous controls, individual evaluation of normalizers for each experiment is imperative for accurate miRNA expression measurements. This approach, which combines multiple techniques and assessments, provides a reliable strategy for identifying and validating endogenous controls in miRNA studies.

• MeSH
• algoritmy MeSH
• mikro RNA * genetika   metabolismus   MeSH
• modely nemocí na zvířatech * MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nealkoholová steatóza jater * genetika   metabolismus   patologie   MeSH
• nitrobřišní tuk * metabolismus   MeSH
• regulace genové exprese MeSH
• stanovení celkové genové exprese metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

We investigated the sex-dependent effects of inflammatory responses in visceral adipose tissue (VAT) and perivascular adipose tissue (PVAT), as well as hematological status, in relation to cardiovascular disorders associated with prediabetes. Using male and female hereditary hypertriglyceridemic (HHTg) rats-a nonobese prediabetic model featuring dyslipidemia, hepatic steatosis, and insulin resistance-we found that HHTg females exhibited more pronounced hypertriglyceridemia than males, while HHTg males had higher non-fasting glucose levels. Additionally, HHTg females had higher platelet counts, larger platelet volumes, and lower antithrombin inhibitory activity. Regarding low-grade chronic inflammation, HHTg males exhibited increased serum leptin and leukocyte levels, while females had increased serum interleukin-6 (IL-6). Both sexes had increased circulating plasminogen activator inhibitor-1 (PAI-1), higher PAI-1 gene expression in VAT and PVAT, and elevated intercellular adhesion molecule-1 (ICAM-1) gene expression in the aorta, contributing to endothelial dysfunction in the HHTg strain. However, HHTg females had lower tumor necrosis factor alpha (TNFα) gene expression in the aorta. Severe dyslipidemia in this prediabetic model was associated with hypercoagulation and low-grade chronic inflammation. The increase in PAI-1 expression in both VAT and PVAT seems to indicate a link between inflammation and vascular dysfunction. Despite the more pronounced dyslipidemia and procoagulation status in females, their milder inflammatory response may reflect an association between reduced cardiovascular damage and prediabetes.

• MeSH
• dyslipidemie * metabolismus   patologie   genetika   MeSH
• inhibitor aktivátoru plazminogenu 1 * metabolismus   genetika   MeSH
• krysa rodu rattus MeSH
• modely nemocí na zvířatech * MeSH
• nemoci cév metabolismus   patologie   etiologie   genetika   MeSH
• nitrobřišní tuk * metabolismus   patologie   MeSH
• pohlavní dimorfismus MeSH
• zánět * metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: This study tested the hypothesis that limited subcutaneous adipose tissue (SAT) expansion represents a primary predisposition to the development of type 2 diabetes mellitus (T2DM), independent of obesity, and identified novel markers of SAT dysfunction in the inheritance of T2DM. METHODS: First-degree relatives (FDR) of T2DM patients (n = 19) and control individuals (n = 19) without obesity (fat mass < 25%) were cross-sectionally compared. Body composition (bioimpedance, computed tomography) and insulin sensitivity (IS; oral glucose tolerance test, clamp) were measured. SAT obtained by needle biopsy was used to analyze adipocyte size, lipidome, mRNA expression, and inflammatory markers. Primary cultures of adipose precursors were analyzed for adipogenic capacity and metabolism. RESULTS: Compared with control individuals, FDR individuals had lower IS and a higher amount of visceral fat. However, SAT-derived adipose precursors did not differ in their ability to proliferate and differentiate or in metabolic parameters (lipolysis, mitochondrial oxidation). In SAT of FDR individuals, lipidomic and mRNA expression analysis revealed accumulation of triglycerides containing polyunsaturated fatty acids and increased mRNA expression of lysyl oxidase (LOX). These parameters correlated with IS, visceral fat accumulation, and mRNA expression of inflammatory and cellular stress genes. CONCLUSIONS: The intrinsic adipogenic potential of SAT is not affected by a family history of T2DM. However, alterations in LOX mRNA and polyunsaturated fatty acids in triacylglycerols are likely related to the risk of developing T2DM independent of obesity.

• MeSH
• diabetes mellitus 2. typu * genetika   metabolismus   MeSH
• inzulinová rezistence * genetika   MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• nenasycené mastné kyseliny metabolismus   MeSH
• nitrobřišní tuk metabolismus   MeSH
• obezita genetika   metabolismus   MeSH
• podkožní tuk metabolismus   MeSH
• průřezové studie MeSH
• triglyceridy metabolismus   MeSH
• tuková tkáň metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The aim of this study was to monitor changes in the components of the metabolic syndrome defined by Adult Treatment Panel III and the risk of adipose tissue. The study population consisted of 45 patients (30 women, 15 men) who underwent one bariatric procedure - partial jejuno-ileal derivation (n=17), sleeve resection (n=14) or laparoscopic gastric - plication (n=14). Components of metabolic syndrome such as waist circumference, morning glycemia/antihypertension, TAG, HDL cholesterol and blood pressure (BP)/antihypertension were monitored in probands. In addition, Dual Energy X-Ray Absorciometry measurements were performed. Parameters were monitored over the course of one year. The study shows that it is an effective method of weight reduction for the study population with metabolic effects in the risk components of metabolic syndrome - fasting glycemia, increase in HDL cholesterol and reduction in triacylglycerols in the blood, reduction in waist circumference and BP or direct disappearance of metabolic syndrome. Significantly, of the entire cohort, 68.9 % of the probands studied showed signs of metabolic syndrome when measured before the intervention. At the end of follow-up, only 22.2 % of probands showed metabolic syndrome. It was also found that if the amount of visceral fat was reduced, the overall risk of metabolic syndrome was also reduced. The study demonstrates a significant positive effect of bariatric surgery on parameters of metabolic syndrome. The study also showed a positive effect of reduced visceral fat volume on the components of metabolic syndrome.

• MeSH
• bariatrická chirurgie * MeSH
• dospělí MeSH
• HDL-cholesterol MeSH
• index tělesné hmotnosti MeSH
• lidé MeSH
• metabolický syndrom * MeSH
• nitrobřišní tuk metabolismus   MeSH
• pilotní projekty MeSH
• tuková tkáň metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

In this study, we aimed to comprehensively characterize the proteomic landscapes of subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) in patients with severe obesity, to establish their associations with clinical characteristics, and to identify potential serum protein biomarkers indicative of tissue-specific alterations or metabolic states. We conducted a cross-sectional analysis of 32 patients with severe obesity (16 males and 16 females) of Central European descent who underwent bariatric surgery. Clinical parameters and body composition were assessed using dual-energy X-ray absorptiometry (DXA) and bioelectrical impedance, with 15 patients diagnosed with type 2 diabetes (T2D) and 17 with hypertension. Paired SAT and VAT samples, along with serum samples, were subjected to state-of-the-art proteomics liquid chromatography-mass spectrometry (LC-MS). Our analysis identified 7,284 proteins across SAT and VAT, with 1,249 differentially expressed proteins between the tissues and 1,206 proteins identified in serum. Correlation analyses between differential protein expression and clinical traits suggest a significant role of SAT in the pathogenesis of obesity and related metabolic complications. Specifically, the SAT proteomic profile revealed marked alterations in metabolic pathways and processes contributing to tissue fibrosis and inflammation. Although we do not establish a definitive causal relationship, it appears that VAT might respond to SAT metabolic dysfunction by potentially enhancing mitochondrial activity and expanding its capacity. However, when this adaptive response is exceeded, it could possibly contribute to insulin resistance (IR) and in some cases, it may be associated with the progression to T2D. Our findings provide critical insights into the molecular foundations of SAT and VAT in obesity and may inform the development of targeted therapeutic strategies.NEW & NOTEWORTHY This study provides insights into distinct proteomic profiles of subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT), and serum in patients with severe obesity and their associations with clinical traits and body composition. It underscores SAT's crucial role in obesity development and related complications, such as insulin resistance (IR) and type 2 diabetes (T2D). Our findings emphasize the importance of understanding the SAT and VAT balance in energy homeostasis, proteostasis, and the potential role of SAT capacity in the development of metabolic disorders.

• MeSH
• biologické markery metabolismus   MeSH
• diabetes mellitus 2. typu * metabolismus   MeSH
• inzulinová rezistence * MeSH
• lidé MeSH
• morbidní obezita * metabolismus   MeSH
• nitrobřišní tuk metabolismus   MeSH
• obezita metabolismus   MeSH
• podkožní tuk metabolismus   MeSH
• proteiny metabolismus   MeSH
• proteomika MeSH
• průřezové studie MeSH
• tuková tkáň metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

INTRODUCTION: Pancreatic steatosis (PS) has both metabolic consequences and local effects on the pancreas itself. Magnetic resonance imaging (MRI) is the most reliable non-invasive method for diagnosing PS. We investigated the impact of metabolic syndrome (MS) on the presence of PS, differences in individuals with and without PS, and the metabolic effects of bariatric procedures. METHODS: Changes in anthropometric and basic biochemistry values and MS occurrence were evaluated in 34 patients with obesity who underwent a bariatric procedure. After the procedure, patients underwent MRI with manual 3D segmentation mask creation to determine the pancreatic fat content (PFC). We compared the differences in the PFC and the presence of PS in individuals with and without MS and compared patients with and without PS. RESULTS: We found no significant difference in the PFC between the groups with and without MS or in the occurrence of PS. There were significant differences in patients with and without PS, especially in body mass index (BMI), fat mass, visceral adipose tissue (VAT), select adipocytokines, and lipid spectrum with no difference in glycemia levels. Significant metabolic effects of bariatric procedures were observed. CONCLUSIONS: Bariatric procedures can be considered effective in the treatment of obesity, MS, and some of its components. Measuring PFC using MRI did not show any difference in relation to MS, but patients who lost weight to BMI < 30 did not suffer from PS and had lower overall fat mass and VAT. Glycemia levels did not have an impact on the presence of PS.

• MeSH
• bariatrická chirurgie * MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• metabolický syndrom * diagnostické zobrazování   metabolismus   MeSH
• morbidní obezita * chirurgie   MeSH
• nitrobřišní tuk metabolismus   MeSH
• obezita metabolismus   MeSH
• pankreas diagnostické zobrazování   metabolismus   MeSH
• retrospektivní studie MeSH
• ztučnělá játra * patologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

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

Schizophrenia is a severe neuropsychiatric disease associated with substantially higher mortality. Reduced life expectancy in schizophrenia relates to an increased prevalence of metabolic disturbance, and antipsychotic medication is a major contributor. Molecular mechanisms underlying adverse metabolic effects of antipsychotics are not fully understood; however, adipose tissue homeostasis deregulation appears to be a critical factor. We employed mass spectrometry-based untargeted proteomics to assess the effect of chronic olanzapine, risperidone, and haloperidol treatment in visceral adipose tissue of prenatally methylazoxymethanol (MAM) acetate exposed rats, a well-validated neurodevelopmental animal model of schizophrenia. Bioinformatics analysis of differentially expressed proteins was performed to highlight the pathways affected by MAM and the antipsychotics treatment. MAM model was associated with the deregulation of the TOR (target of rapamycin) signalling pathway. Notably, alterations in protein expression triggered by antipsychotics were observed only in schizophrenia-like MAM animals where we revealed hundreds of affected proteins according to our two-fold threshold, but not in control animals. Treatments with all antipsychotics in MAM rats resulted in the downregulation of mRNA processing and splicing, while drug-specific effects included among others upregulation of insulin resistance (olanzapine), upregulation of fatty acid metabolism (risperidone), and upregulation of nucleic acid metabolism (haloperidol). Our data indicate that deregulation of several energetic and metabolic pathways in adipose tissue is associated with APs administration and is prominent in MAM schizophrenia-like model but not in control animals.

• MeSH
• antipsychotika terapeutické užití   MeSH
• haloperidol farmakologie   terapeutické užití   MeSH
• krysa rodu rattus MeSH
• methylazoxymethanolacetát farmakologie   MeSH
• modely nemocí na zvířatech MeSH
• nitrobřišní tuk účinky léků   embryologie   metabolismus   MeSH
• olanzapin farmakologie   terapeutické užití   MeSH
• potkani Sprague-Dawley MeSH
• proteomika MeSH
• risperidon farmakologie   terapeutické užití   MeSH
• schizofrenie farmakoterapie   MeSH
• signální transdukce účinky léků   MeSH
• těhotenství MeSH
• TOR serin-threoninkinasy metabolismus   MeSH
• tuková tkáň účinky léků   metabolismus   MeSH
• zpožděný efekt prenatální expozice chemicky indukované   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Ovarian hormone deficiency leads to increased body weight, visceral adiposity, fatty liver and disorders associated with menopausal metabolic syndrome. To better understand the underlying mechanisms of these disorders in their early phases of development, we investigated the effect of ovariectomy on lipid and glucose metabolism. Compared to sham-operated controls, ovariectomized Wistar female rats markedly increased whole body and visceral adipose tissue weight (p ˂ 0.05) and exhibited insulin resistance in peripheral tissues. Severe hepatic triglyceride accumulation (p ˂ 0.001) after ovariectomy preceded changes in both serum lipids and glucose intolerance, reflecting alterations in some CYP proteins. Increased CYP2E1 (p ˂ 0.05) and decreased CYP4A (p ˂ 0.001) after ovariectomy reduced fatty acid oxidation and induced hepatic steatosis. Decreased triglyceride metabolism and secretion from the liver contributed to hepatic triglyceride accumulation in response to ovariectomy. In addition, interscapular brown adipose tissue of ovariectomized rats exhibited decreased fatty acid oxidation (p ˂ 0.01), lipogenesis (p ˂ 0.05) and lipolysis (p ˂ 0.05) despite an increase in tissue weight. The results provide evidence that impaired hepatic triglycerides and dysregulation of some CYP450 proteins may have been involved in the development of hepatic steatosis. The low metabolic activity of brown adipose tissue may have contributed to visceral adiposity as well as triglyceride accumulation during the postmenopausal period.

• MeSH
• bílá tuková tkáň metabolismus   MeSH
• dieta s vysokým obsahem tuků MeSH
• dyslipidemie metabolismus   MeSH
• glukosa metabolismus   MeSH
• hmotnostní přírůstek MeSH
• hnědá tuková tkáň metabolismus   MeSH
• inzulin metabolismus   MeSH
• inzulinová rezistence MeSH
• játra metabolismus   MeSH
• krysa rodu rattus MeSH
• lipidy krev   MeSH
• lipogeneze účinky léků   MeSH
• lipolýza MeSH
• menopauza metabolismus   fyziologie   MeSH
• metabolismus lipidů účinky léků   fyziologie   MeSH
• nitrobřišní tuk metabolismus   MeSH
• obezita metabolismus   MeSH
• ovarektomie škodlivé účinky   MeSH
• poruchy metabolismu lipidů etiologie   patofyziologie   MeSH
• postmenopauza metabolismus   fyziologie   MeSH
• potkani Wistar MeSH
• systém (enzymů) cytochromů P-450 metabolismus   fyziologie   MeSH
• triglyceridy metabolismus   MeSH
• ztučnělá játra metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Metabolic syndrome (MetS) is an important cause of worldwide morbidity and mortality. Its complex pathogenesis includes, on the one hand, sedentary lifestyle and high caloric intake, and, on the other hand, there is a clear genetic predisposition. PD (Polydactylous rat) is an animal model of hypertriglyceridemia, insulin resistance, and obesity. To unravel the genetic and pathophysiologic background of this phenotype, we compared morphometric and metabolic parameters as well as liver transcriptomes among PD, spontaneously hypertensive rat, and Brown Norway (BN) strains fed a high-fat diet (HFD). After 4 weeks of HFD, PD rats displayed marked hypertriglyceridemia but without the expected hepatic steatosis. Moreover, the PD strain showed significant weight gain, including increased weight of retroperitoneal and epididymal fat pads, and impaired glucose tolerance. In the liver transcriptome, we found 5480 differentially expressed genes, which were enriched for pathways involved in fatty acid beta and omega oxidation, glucocorticoid metabolism, oxidative stress, complement activation, triacylglycerol and lipid droplets synthesis, focal adhesion, prostaglandin synthesis, interferon signaling, and tricarboxylic acid cycle pathways. Interestingly, the PD strain, contrary to SHR and BN rats, did not express the Acsm3 (acyl-CoA synthetase medium-chain family member 3) gene in the liver. Together, these results suggest disturbances in fatty acid utilization as a molecular mechanism predisposing PD rats to hypertriglyceridemia and fat accumulation.

• MeSH
• dieta s vysokým obsahem tuků škodlivé účinky   MeSH
• exprese genu MeSH
• hypertriglyceridemie krev   genetika   MeSH
• játra metabolismus   MeSH
• koenzym A-ligasy genetika   MeSH
• krysa rodu rattus MeSH
• modely nemocí na zvířatech MeSH
• nitrobřišní tuk metabolismus   MeSH
• polydaktylie MeSH
• potkani inbrední SHR MeSH
• potkani Wistar MeSH
• stanovení celkové genové exprese metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH