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- Sarsenbayeva, Assel, et al.
(författare)
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Excess glucocorticoid exposure contributes to adipose tissue fibrosis which involves macrophage interaction with adipose precursor cells
- 2022
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Ingår i: Biochemical Pharmacology. - : Elsevier. - 0006-2952 .- 1356-1839. ; 198
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Tidskriftsartikel (refereegranskat)abstract
- Chronic exposure to elevated glucocorticoid levels, as seen in patients with Cushing’s syndrome, can induce adipose tissue fibrosis. Macrophages play a pivotal role in adipose tissue remodelling. We used the synthetic glucocorticoid analogue dexamethasone to address glucocorticoid effects on adipose tissue fibrosis, in particular involving macrophage to preadipocyte communication. We analysed the direct effects of dexamethasone at a supra-physiological level, 0.3 µM, on gene expression of pro-fibrotic markers in human subcutaneous adipose tissue. The effects of dexamethasone on the differentiation of human SGBS preadipocytes were assessed in the presence or absence of THP1-macrophages or macrophage-conditioned medium. We measured the expression of different pro-fibrotic factors, including α-smooth muscle actin gene (ACTA2) and protein (α-SMA). Dexamethasone increased the expression of pro-fibrotic genes, e.g. CTGF, COL6A3, FN1, in adipose tissue. Macrophages abolished preadipocyte differentiation and increased the expression of the ACTA2 gene and α-SMA protein in preadipocytes after differentiation. Exposure to dexamethasone during differentiation reduced adipogenesis in preadipocytes, and elevated the expression of pro-fibrotic genes. Moreover, dexamethasone added together with macrophages further increased ACTA2 and α-SMA expression in preadipocytes, making them more myofibroblast-like. Cells differentiated in the presence of conditioned media from macrophages pretreated with or without dexamethasone had a higher expression of profibrotic genes compared to control cells. Our data suggest that macrophages promote adipose tissue fibrosis by directly interfering with preadipocyte differentiation and stimulating gene expression of pro-fibrotic factors. Excess glucocorticoid exposure also has pro-fibrotic effect on adipose tissue, but this requires the presence of macrophages.
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| 2. |
- Sarsenbayeva, Assel, et al.
(författare)
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Human macrophages stimulate expression of inflammatory mediators in adipocytes; effects of second-generation antipsychotics and glucocorticoids on cellular cross-talk
- 2021
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Ingår i: Psychoneuroendocrinology. - : Elsevier. - 0306-4530 .- 1873-3360. ; 125
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Tidskriftsartikel (refereegranskat)abstract
- Objective: Adipose tissue inflammation and distorted macrophage-adipocyte communication are positively associated with metabolic disturbances. Some pharmacological agents, such as second-generation antipsychotics (SGAs) and synthetic glucocorticoid (GC) dexamethasone, tend to induce adverse metabolic side effects and the underlying mechanisms are not fully understood. Our work aimed to study whether SGAs and dexamethasone affect macrophage phenotype and macrophage-adipocyte communication on gene expression level. We selected the model involving THP-1-derived macrophages, polarized into M0, M1, and M2 phenotypes, and primary human mature subcutaneous adipocytes.Methods: Abdominal subcutaneous adipose tissue needle biopsies were obtained from 6 healthy subjects (4F/2M; age: 22-64 yr; BMI: 21.7-27.6 kg/m2) followed by isolation of mature adipocytes. THP-1-human monocytic cell line was used for the study. THP-1 monocytes were differentiated and polarized into M0 (naive), M1 (classically activated), and M2 (alternatively activated) macrophages. During and after polarization the macrophages were treated for 24 h without (control) or with therapeutic and supra-therapeutic concentrations of olanzapine (0.2 mu M and 2.0 mu M), aripiprazole (1.0 mu M and 10 mu M) and its active metabolite dehydroaripiprazole (0.4 mu M and 4.0 mu M). Isolated mature human adipocytes were co-incubated with THP-1-derived polarized macrophages pretreated with SGAs after their polarization. Adipocytes and macrophages were collected before and after co culture for mRNA expression analysis of genes involved in inflammation.Results: Co-incubation of mature human adipocytes with human macrophages, regardless of polarization, resulted in a marked induction of pro-inflammatory cytokines in adipocytes, including IL1B,IL6, TNFA, and IL10. Remarkably, it did not affect the expression of adipokines and genes involved in the regulation of energy, lipid, and glucose metabolism in adipocytes. Dexamethasone markedly reduced gene expression of pro-inflammatory cytokines in macrophages and prevented macrophage-induced inflammatory response in adipocytes. In contrast, SGAs did not affect macrophage-adipocyte communication and had a minute anti-inflammatory effect in macrophages at supra-therapeutic concentrations. Interestingly, the adipocytes co-incubated with M1 macrophages pre-treated with dexamethasone and SGAs particularly the supra-therapeutic concentration of olanzapine, reduced expression of LPL, LIPE, AKT1, and SLC2A4, suggesting that the expression of metabolic genes in adipocytes was dependent on the presence of pro-inflammatory M1 macrophages.Conclusion: Together, these data suggest that macrophages induce expression of pro-inflammatory genes in human subcutaneous adipocytes without affecting the expression of adipokines or genes involved in energy regulation. Furthermore, our findings demonstrated that SGAs and dexamethasone had a mild effect on macrophage-adipocyte communication in M1 macrophage phenotype.
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| 3. |
- Sarsenbayeva, Assel, et al.
(författare)
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Role of glucocorticoids in adipose tissue fibrosis and interplay between macrophages and adipose cells
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Background and aimsExcessive endogenous production and administration of glucocorticoids leads to adipose tissue fibrosis as well as metabolic complications. Alternatively activated M2 macrophages are known to interfere with adipocyte differentiation and to promote phenotypic switch of preadipocytes into pro-fibrotic myofibroblasts. Glucocorticoids are known to promote M2 phenotype in macrophages. Therefore, the aim of this study was to investigate whether the effects of synthetic glucocorticoid dexamethasone on adipose tissue fibrosis are mediated through on macrophages to adipocyte communication. MethodsTranscriptomics analysis was performed on human adipose tissue treated without and with dexamethasone. We tested the effects of dexamethasone on differentiation of human SGBS adipocyte cells in the absence or presence of macrophages derived from THP-1 monocytic cell line plated on inserts. The differentiation rate was assessed by measuring cell lipid accumulation, expression of markers of adipogenesis PPARG and CEBPA. Acquisition of myofibroblast phenotype by preadipocytes was assessed by measuring the expression of myofibroblast marker α-smooth muscle actin. The pro-fibrotic activity of these cells was measured by the expression of collagen VI.ResultsOur transcriptomics data demonstrated that dexamethasone is able to directly double the expression of pro-fibrotic genes, such as CTGF, COL6A3, FN1, in adipose tissue, compared to control (p<0.05). Expression of CD163, a marker of M2 macrophages, positively correlated with the components of extracellular matrix COL6A3 and FN1 (p<0.01), and with the pro-fibrotic genes CTGF and AXL (p<0.01) in human adipose tissue. In addition, dexamethasone induced the expression of CD163 and MRC1 in THP-1 macrophages (p<0.05), suggesting that dexamethasone drives M2 phenotype in macrophages. We observed that dexamethasone inhibited adipogenesis by ~30% (p<0.01). Macrophages almost completely abolished differentiation of adipocytes by ~90% (p<0.01). Additionally, macrophages induced 2-3 fold increase in the expression of ACTA2 gene and protein in adipocytes (p<0.01). Dexamethasone alone did not affect ACTA2 in adipocytes,but in the presence of macrophages, increased ACTA2, when compared to macrophages alone (p<0.05). A similar nominal increase was observed in COL6A3. ConclusionOur data show that dexamethasone has a direct pro-fibrotic effect on adipose tissue and promotes M2 phenotype in macrophages. Furthermore, dexamethasone potentiated macrophage-induced phenotype switch of adipocytes to myofibroblasts, which suggests that its effect on adipose tissue fibrosis is mediated though macrophage-adipocyte communication.
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