Increased Circulating Angiopoietin-Like Protein 8 Levels Are Associated with Thoracic Aortic Dissection and Higher Inflammatory Conditions

Figures

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  Fig. 1. Receiving operator curves using ANGPTL8 and hs-CRP plasma levels for prediction of TAD. The area under the curve (AUC) for the plasma ANGPTL8 levels was 0.746, for hs-CRP levels was 0.807, for D-dimer was 0.877, and for the combination of ANGPTL8, hs-CRP, and D-dimer was 0.927

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  Fig. 2. ANGPTL8 is increased in thoracic aortic dissection (TAD) tissues and expressed in macrophages and vascular smooth cells. a Representative immunostaining and semiquantitative analysis of ANGPTL8 in the aortas of TAD and control patients (n = 6 per group). Data are presented as mean ± standard error of the mean (SEM). *P < 0.05. b Co-staining of α-SMA (red) and ANGPTL8 (green) in TAD tissues. c Co-staining of MAC-2 (red) and ANGPTL8 (green) in TAD tissues. Data are presented as mean ± SEM. *P < 0.05

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  Fig. 3. ANGPTL8 expression was increased in angiotensin II (AngII)-induced macrophage cells, while inhibition of ANGPTL8 via ANGPTL8 siRNA decreased the expression of ANGPTL8 and inflammatory factors. a, b ANGPTL8 levels in RAW264.7 cells were increased after AngII treatment, as assessed by Western blot. c, d ANGPTL8 siRNA decreased the expression of ANGPTL8 in RAW264.7 cells induced by AngII. e RAW264.7 cells were treated with 25, 50, or 100 nmol/L AngII for 24 h. The 100 nmol/L AngII was used for the ANGPTL8 siRNA study. f–i qPCR analysis of gene expression of ANGPTL8 and the inflammatory factors TNF-α, IL-6, MMP-9, and IL-1β in RAW264.7 cells. j–m ELISA analysis of macrophage secreted IL-1β, IL-6, TNF-α, and MCP-1. The RAW264.7 cells were treated with AngII (100 nmol/L) except for the control group. ANGPTL8 siRNA-1 and ANGPTL8 siRNA-2 represent the two sequences of ANGPTL8 siRNA. *P < 0.05 vs. control group, #P < 0.05 vs. AngII group. ANGPTL8 siRNA decreased the expression of the inflammatory factors induced by AngII in RAW264.7 cells. Data are presented as mean ± SEM. *P < 0.05

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  Fig. 4. ANGPTL8 expression was increased in angiotensin II (AngII)-induced HUASMCs, while inhibition of ANGPTL8 via ANGPTL8 siRNA decreased the expression of ANGPTL8 and apoptosis markers. a, b ANGPTL8 levels in HUASMCs were increased after AngII treatment, as assessed by Western blot. c, d ANGPTL8 siRNA decreased the expression of ANGPTL8 in HUASMCs induced by AngII. e HUASMCs were treated with 25, 50, or 100 nmol/L AngII for 24 h. The 100 nmol/L AngII was used for the ANGPTL8 siRNA study. f, g qPCR analysis of the gene expression of ANGPTL8 and the antiapoptotic genes Bcl-2 and Bim in HUASMCs. h, i TUNEL staining to detect the apoptosis of HUASMCs induced by AngII. The HUASMCs were treated with AngII (100 nmol/L) except for the control group. ANGPTL8 siRNA-1 and ANGPTL8 siRNA-2 represent the two sequences of ANGPTL8 siRNA *P < 0.05 vs. control group, #P < 0.05 vs. AngII group. ANGPTL8 siRNA decreased the expression of the apoptotic factors and decreased the apoptosis of HUASMCs induced by AngII. Data are presented as mean ± SEM. *P < 0.05