Omentin-1 is associated with atrial fibrillation in patients with cardiac valve disease

Figures

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  Fig. 1. Epicardial adipose tissue (EAT) structure, omentin-1, and TGF-β1 expression in human samples. Representative pictures of Masson’s trichrome stained right atrial appendages (a) (100× magnification) revealed severe atrial fibrosis in patients with atrial fibrillation (AF) (AF group, n = 18; nAF group, n = 13). Representative immunohistochemical images of omentin-1 (b) in right atrial appendages and quantitative protein expression level. Omentin-1 expression in right atrial appendages was detected via western blotting (c) (AF group, n = 4; nAF group, n = 4). Representative images of H&E-stained EAT (d) (100× magnification) showed adipocyte hypertrophy in patients with AF (AF group, n = 20; nAF group, n = 20). Representative images of Masson’s trichrome-stained EAT (D) (100× magnification) indicated severe EAT fibrosis in patients with AF. Representative immunohistochemical images of omentin-1 (e) and TGF-β1 (e) in EAT and respective quantitative protein expression levels. *P < 0.05 vs nAF group, **P < 0.01 vs nAF group, ***P < 0.001 vs nAF group

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  Fig. 2. Omentin-1 inhibited TGF-β1-induced activation of cardiac fibroblasts (CFs). α-SMA, COL1, and COL3 protein levels in non-treated and treated CFs were detected via western blotting (a). The mRNA levels of α-SMA, COL1a, and COL3a1were detected via RT-qPCR. Expression of mRNA was normalized to GAPDH. (b) Representative images of immunofluorescence staining for α-SMA in CFs (c) (200× magnification) showed the antifibrotic effect of omentin-1. p-SMAD2, t-SMAD2, p-SMAD3, t-SMAD3 protein levels in CFs were detected via western blotting (d). (e) CF proliferation assay. (f) Representative images of the CFs scratch assay (100× magnification). The values were mean ± SEM of three independent experiments. *P < 0.05 vs control group, **P < 0.01 vs control group, ***P < 0.001 vs control group, †P < 0.05 vs TGF-β1 group, ††P < 0.01 vs TGF-β1 group, †††P < 0.001 vs TGF-β1 group

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  Fig. 3. Omentin-1 inhibited TGF-β1-induced endothelial-mesenchymal transition (EndMT) of human umbilical vein endothelial cells (HUVECs). Vimentin and VE-Cad protein levels in HUVECs were detected via western blotting (a). The mRNA levels of vimentin and VE-Cad were detected via RT-qPCR (b). Expression of mRNA were normalized to GAPDH. Representative images of immunofluorescence staining for vimentin (c) and VE-Cad (d) (200× magnification) in HUVECs. The p-SMAD3 and t-SMAD3 protein levels in HUVECs were detected via western blotting (e). (f) Representative images of HUVECs scratch assay (× 100 magnification). The values were mean ± SEM of three independent experiments. *P < 0.05 vs control group, **P < 0.01 vs control group, ***P < 0.001 vs control group, †P < 0.05 vs TGF-β1 group, ††P < 0.01 vs TGF-β1 group, †††P < 0.001 vs TGF-β1 group

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  Fig. 4. Hypoxia resulted in a change in the secretion phenotype of adipocytes. Omentin-1 protein levels in adipocytes was detected via western blotting (a). The concentrations of omentin-1 in the CM of adipocyte treated with normoxia or hypoxia were determined by ELISA (b). The mRNA levels of TGF-β1 in adipocytes (24 h normoxia, 24 h hypoxia, 36 h normoxia, 36 h hypoxia) was detected via RT-qPCR. Expression of mRNA was normalized to GAPDH. (c) The mRNA levels of α-SMA, COL1a, and COL3a1 in non-CM-treated or CM-treated CFs were detected via RT-qPCR. The mRNA level was normalized to GAPDH. (d, e) (f) Representative images of CFs scratch assay (100× magnification). The values were mean ± SEM of three independent experiments. *P < 0.05 vs normoxia 24 h group, ***P < 0.001 vs normoxia 24 h group, #P < 0.05 vs normoxia 36 h group, ###P < 0.001 vs normoxia 36 h group, †P < 0.05 vs control group, ††P < 0.01 vs control group, †††P < 0.001 vs control group