Fig. 1
Glucose or endocrine activity of adipose tissue after Semaglutide treatment. A Workflow of EAT and SAT treatment with semaglutide (Sema) (1 nM), insulin (INS) (2.5 or 5 μg/mL) or both for 90 min. Afterwards, glucose was analyzed by colorimetric assay or exosomes were isolated by ExoSpin kit for proteomics analysis. B Bar plot shows mean ± SD of GLP1 receptor (NM_002062.5) mRNA expression levels on epicardial and subcutaneous adipose tissue (EAT and SAT) from patients underwent cardiac surgery (n = 2). Their mRNA expression levels were quantified by quantitative PCR and represented according to β-actin (NM_001101.5) expression levels as arbitrary units (a.u.) based on 2−(ACTB/gene) algorithm. Different regions were checked. Bar plots shows mean ± SD of glucose uptake by EAT and SAT under different treatments and times. Glucose consumption by SAT after insulin treatment was performed for 30 and 90 min (n = 2), glucose consumption by EAT and SAT from 3 patients was analyzed after insulin treatment at different concentrations (2.5 and 5 μg/mL). Wilcoxon rank test determined unsignificant changes. C 9 out of 12 SAT or EAT samples were insulin responders. Dot plot shows individual data, mean ± SD of glucose consumption by EAT and SAT after semaglutide (Sema), insulin (INS) or combined treatment (Sema + INS). Mann–Whitney test did not show statistical significance regarding EAT. Nonetheless, the glucose consumption increased after insulin [n = 9, CONTROL (15.590 ± 8.053) vs INS (24.240 ± 7.332), *p = .0400] and combined treatment [n = 5, CONTROL (15.590 ± 8.053) vs Sema + INS (29.992 ± 6.243), **p = .0040]. Bar plot represents mean ± SD of glucose consumption (mg/dL) after fat biopsies treatment in those selected patients for proteomics analysis (n = 3). D Venn’s diagram and String plots of the identified proteins from exosomes by TripleTOF. Proteins on each group of treatment were selected if they were identified at least twice. They were included on FunRich software for identifying those differential proteins regarding treatments. A1TR (SERPINA1), CUL7, KV37 and TRPV5 were only identified after insulin (INS) treatment on EAT. STRING analysis shows a network among A1TR, CUL7 and INS. GELS (GSN) and TFRE were identified after Semaglutide (Sema) treatment. STRING analysis showed relationship with GLP1R. On subcutaneous fat (SAT), PEDF, CO4B, K2C5, KV320, PRDX1, ANXA5, GELS (GSN) and FABP4 were only identified after cotreatment (Sema and INS). STRING analysis shows a network among the last three proteins (ANXA5, GSN and FABP4), INS and GLP1R. E Upper left representative western blot of epicardial (EAT) and subcutaneous fat (SAT) proteins from exosomes and eluted part or total secretome with (+) or without (−) washing with saline solution (PBS) and stained with albumin and CD9 tetraspanin antibodies. Upper right bar graph represents mean ± SD of FABP4 intensity identified on following western blots of SAT with combined treatment (Sema + INS) regarding control from 7 patients. Wilcoxon test did not show statistical significance. Down representative western blots of SAT with FABP4 or IgG antibodies (Ab)