PFOS and Its Substitute OBS Cause Endothelial Dysfunction to Promote Atherogenesis in ApoE –/– Mice

Figures

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  Figure 1. Effects of PFOS and OBS on the levels of blood lipids in ApoE–/– mice. (A) Flowchart of animal experimental procedure. (B–E) Levels of TG, TC, HDL-C, and LDL-C in plasma of mice at 6, 8, and 12 weeks postexposure (n = 4). (F) Levels of PFOS and OBS in plasma of mice at 12 weeks postexposure (n = 6). (G) Mass spectrometry images show the spatial distribution of PFOS in the aorta of mice, and average fluorescence intensity was quantitatively analyzed (n = 3). *P < 0.05, **P < 0.01. Scale bar: 100 μm.

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  Figure 2. Effects of PFOS and OBS on vascular endothelium in the aorta of ApoE–/– mice. (A,B) Fluorescent images (A) and quantitative analysis (B) of VE-cadherin protein expression in aortic tissues of mice at 12 weeks after PFOS or OBS exposure. (C–E) Relative expression levels of cell adhesion molecules (C), inflammatory factors (D) and cell junction genes (E) determined using RT-qPCR in aortic tissues of mice at 12 weeks after PFOS or OBS exposure (n = 4). *P < 0.05, **P < 0.01. Scale bar: 100 μm.

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  Figure 3. Effects of exposure to OBS on endothelial cytotoxicity and gene expression profile in HUVECs. (A-D) Cells were exposed to PFOS or an OBS for 24 and 48 h, followed by cell viability (A and B) and LDH release assays (C and D). (E) Volcano plots showed the changes in mRNA expression levels after cells were exposed to 20 μmol/L OBS for 48 h. (F) GO analysis of significantly upregulated genes was shown. (G) KEGG analysis of significantly upregulated genes was shown. n = 3, *P < 0.05, **P < 0.01.

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  Figure 4. Impacts of PFOS and OBS on cell adhesion and inflammation in HUVECs. (A and B) Cell adhesion of THP-1 cells labeled with BCCF-AM to HUVECs were observed and the number of adhering cells were counted. (C–F) Representative Western blots were shown and protein expression levels of cell adhesion molecules and inflammation factors were quantified after exposure to PFOS (C and E) and OBS (D and F). n = 3, *P < 0.05, **P < 0.01. Scale bar: 100 μm.

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  Figure 5. Effects of PFOS and OBS exposure on cell permeability and cell junctions in HUVECs. (A) Relative quantitative analysis of endothelial permeability after exposure to PFOS or OBS. (B) Representative fluorescence images of the gap junction assay, and relative distances of scrape after exposure to PFOS or OBS. (C and D) Representative Western blots, and relative expression levels of cell junction proteins after exposure to PFOS (C) or OBS (D). n = 3, *P < 0.05, **P < 0.01. Scale bar: 100 μm.