Upregulated Autophagy in Calcific Aortic Valve Stenosis Confers Protection of Valvular Interstitial Cells

Figures

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  Figure 1. Autophagy pathways are differentially regulated in the calcified tissue of human valves. Gene expression analyses of key autophagy genes in the healthy, thickened, and calcified tissue of human valves from calcific aortic valve stenosis (CAVS; n = 55) and aortic regurgitation (AR; n = 9) patients. Data presented as box-and-whisker plots with 5–95 percentiles. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.

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  Figure 2. Lysosomal pathways are differentially regulated in the calcified tissue of human valves. Gene expression of key lysosomal genes in the healthy, thickened, and calcified tissue of human valves from calcific aortic valve stenosis (CAVS; n = 55) and aortic regurgitation (AR; n = 9) patients. Data presented as box-and-whisker plots with 5–95 percentiles. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.

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  Figure 3. Autophagic activity is differentially expressed in AR and in CAVS and serves as a survival mechanism in valvular interstitial cells (VICs). (A) Immunoblotting against LC3-I and LC3-II in human VICs treated for 2 h with bafilomycin (1nM) (B+) or vehicle control (DMSO) (B-), including quantification of the autophagic flux measured as the LC3-II ratio between (B+/B-)/β-actin. CAVS: n = 5, AR: n = 4. (B) VIC viability from CAVS patients after 24 h treatment with bafilomycin (1nM) or vehicle control (DMSO) n = 7. (C) Immunoblotting against LC3-I and LC3-II in human VICs from CAVS patients treated for 2 h with bafilomycin (1nM) or vehicle control (DMSO) followed by 24 h treatment with 2.6 mM phosphate (Pi) or control (media) n = 4. Quantified as the densiometry of LC3-II/β-actin. Data presented as mean ± S.D. * = p < 0.05, ** = p <0.01, *** = p < 0.001, **** = p < 0.0001.