Research Paper Advance Articles

Figure 1. Functional evaluation of aortic stiffness between old (n = 5–10, red) and young (n = 5–10, blue) mice. (A) Decision tree allowing the functional evaluation of the animal with the elastic fiber. “^*” identifies parameters that are methodologically accessible for clinical studies. (B) mRNA expression of vascular contraction markers (αSMA, Alpha Smooth Muscle Actin - SM22α, Smooth muscle protein 22-alpha - MLCK, Myosin light-chain kinase). (C) mRNA expression of endothelin (ET-1) and endothelial (eNOS) and inducible (iNOS) nitric oxide synthase. (D) Plasma assay of endothelin 1. (E) Ex vivo measurement of vascular reactivity after stimulation with phenylephrine or acetylcholine. (F, G) Western blot of native and phosphorylated forms of eNOS (panel F) and quantification of gray level by ImageJ (panel G). (H) Plasma dosage of nitric oxide (NO). (I, J) Evaluation of extracellular matrix: by atomic force microscopy. (I) Imaging of elastic fibers (EF) with PeakForce error and DMT modulus (scale bar: 10 µm). (J) Quantifications of EF rugosity (left), stiffening of EF (middle) and inter-EF spaces (IEFS, right). Statistical test: Mann-Whitney. Mean+/− SEM. Significant differences (^*p < 0.05, ^**p < 0.001, ^***p < 0.0001, Mann-Whitney).