Fig. 4.

Effects of nAChRs inhibition on PA reactivity. (A) Vascular responses were analyzed in endothelium-intact PAs from C57BL/6J mice (n = 4 animals) from untreated (CTRL), (n = 6 rings), or overnight treated with 15 % CSE (n = 6 rings), alone or in combination with 10 μM Me (Me + CSE, n = 6 rings). Average values of the endothelium-dependent vasodilation induced by increasing amounts of ACh (left) or increasing amounts of riociguat (right) were assessed and expressed as percentages of relaxation related to the maximum 5-HT-driven vasoconstriction. (B) PAs from α7^−/− C57BL/6J mice (n = 5 animals) were untreated (n = 7 rings) or overnight treated with 15 % CSE (n = 6 rings) and vascular responses were analyzed as in (A). (C) C57BL/6J mice (n = 5 per group) were exposed to air (CTRL; n = 8 rings), cigarette smoke (CS; n = 9 rings), or a combination of cigarette smoke and 2 mg/kg Me (Me + CS; n = 9 rings) over a two-week period, following the specified timeline. Afterwards vascular responses were analyzed as in (A). Lung sections from each animal group were stained with nuclear fast red and analyzed by light microscopy. Representative images from controls, CS treated or Me + CS were selected. Data are represented as the mean ± s.e.m.; Statistical comparison between groups was performed using one-way ANOVA test followed by Tukey's test (A, C) or two-sided unpaired Student's t-test (B) (∗)p < 0.05, (∗∗)p < 0.01, (∗∗∗)p < 0.001 (∗∗∗∗)p < 0.0001.