FIGURE 6.

TMAO induced cardiac fibrosis via NLRP3 activation. Knockdown of NLRP3 with siRNA could reverse the effect of TMAO on cardiac fibroblasts. (A) NLRP3, pro-IL-1β, IL-1β, TGF-β, Col I and Col III protein expression levels were analyzed by western blotting and quantified (B) according to immunoblotting (the ratio of protein pixel density/GAPDH pixel density). (C) Representative images and quantification (D) of immunofluorescence staining for α-SMA (red) and DAPI (blue) in cardiac sections. (E) Edu staining showing cardiac fibroblast proliferation after 24 h of treatment, and quantification (F) of the percentage of Edu-positive cells (red). (G) Transwell assay showing the stained migrated cardiac fibroblasts after 24 h and calculation (H). All scale bars = 50 μm. All data were analyzed at least three times. Mean ± SD. ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001 (vs. TMAO+si NC).