NAMPT/SIRT1 Attenuate Ang II-Induced Vascular Remodeling and Vulnerability to Hypertension by Inhibiting the ROS/MAPK Pathway

Figure 3

Ang II-induced endothelial dysfunction and changes in the proliferation and migration of MOVAs are ameliorated by the overexpression of NAMPT. (a) The protein levels and densitometric quantification of apoptosis markers (Caspase 3 and Bcl-2) and P-eNOS/eNOS were determined in cells transfected with the NAMPT plasmid and treated with Ang II for 48 h using Western blot analyses. (b) The expression of the endothelial cell permeability marker ZO-1 in MAECs was determined using immunofluorescence staining (ZO-1: red; nuclei (DAPI): blue). Scale  μm. (c) MAECs subjected to different treatments were stained with Annexin V and propidium iodide (PI) and analyzed using flow cytometry to evaluate the effect of NAMPT on cell apoptosis after the Ang II treatment. Quantification of cell apoptosis is showed in the right panel. (d) Relative levels of proliferation-related proteins (PCNA and cyclin D1) and extracellular matrix synthesis-related proteins (MMP-2 and MMP-9) in cultured MOVAs from the indicated groups. (e, f). Representative images of scratch-wound assays and transwell assays and quantification of migrated MOVAs ( in each group; scale  μm). Data are presented as ., , and compared with the Ctrl or between the two indicated groups.

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