Fig. 3

Circulating NAFLD hepatic sEV induces NLRP3 inflammasome-dependent endothelial hyperpermeability in coronary microvessels. NAFLD or control hepatic sEVs were isolated by differential ultracentrifugation, identified, and administered to naive NLRP3^+/+ and NLRP3^−/− mice via caudal vein injection. A Representative electron micrograph of sEVs reveals the morphology and size. Scale bar, 200 nm. B Size distribution analysis of sEVs by nanoparticle tracking analysis. C Western blot analyses of sEV markers, including CD63, HSP70, and TSG101. D In vivo optical imaging system-obtained fluorescence imaging of cardiac tissue, n = 6 per group. The pellet derived from the ultracentrifugation of DiR alone was used as a vehicle control. E–H Representative fluorescent confocal images of cleaved-caspase-1(CASP1), IL-1β, and ZO-1/2 (green) with vWF (red) and the summarized data of the Manders overlap coefficient. The area of interest (AOI) is selected for higher magnification, n = 6 per group. Scale bar, 20 μm. I, J Representative images and the summarized data of Evans blue concentrations in heart tissues, n = 6 per group. Data are expressed as the mean ± SEM. Statistics: One-way ANOVA, *P < 0.05, **P < 0.01 vs. NLRP3^+/+ mice injected with MCS hepatic sEVs; ^#P < 0.05, ^##P < 0.01 vs. NLRP3^+/+ mice injected with MCD hepatic sEVs. ^&&P < 0.01 vs. NLRP3^+/+ mice injected with ND hepatic sEVs; ^$$P < 0.01 vs. NLRP3^+/+ mice injected with HFD hepatic sEVs