Fig. 8
ENOblock reverses SuHx-induced PH in rats. a A diagrammatic sketch depicts the SuHx-induced PH rats model and ENOblock treatment protocol (n = 6–10 per group). Male SD rats received a dose of Sugen 5416 (20 mg/kg) subcutaneously at the first day of hypoxia exposure (10% O2). After 3-week chronic hypoxia exposure, the rats were placed back to room air for 2 weeks of reoxygenation before the ENOblock treatment. During the treatment period, the rats received 20 mg/kg ENOblock treatment via intraperitoneal injection for 2 weeks (one injection per day in the first week and one injection every other day in the second week). After the completion of ENOblock treatment, we measured RVP and quantified RVSP b (n = 6–10 per group, **P < 0.01), analyzed right ventricular hypertrophy c (n = 6–10 per group, **P < 0.01, *P < 0.05), and assessed pulmonary artery remodeling d (**P < 0.01) were analyzed and calculated accordingly. Representative HE staining images of the pulmonary arteries in the lung sections of experimental rats are shown in e (Scale bars, 100 μm). f The lung sections of experimental rats from each group were immunostained with SMA and Ki67 antibodies and DAPI to visualize the proliferating PASMC (Scale bars, 50 μm) and g the proliferation rates of PASMCs were quantified in each group. h The rat lung sections were co-immunostained with p-ACC and SMA antibodies and DAPI to visualize the AMPK activation in PASMC. The white arrows indicate the PASMC showing high p-ACC signal (Scale bars, 50 μm). i The rat lung sections were co-immunostained with p-PRAS40 and SMA antibodies and DAPI to visualize the Akt activation in PASMC. The white arrows indicate the PASMC showing high p-PRAS40 signal (Scale bars, 50 μm). j A schematic diagram depicts the role of the ENO1-AMPK-Akt axis in PH. ENO1 activates the AMPKα-Akt signaling cascade, therefore promotes PASMC proliferation, de-differentiation, apoptotic resistance, and metabolic shift, leading to PH. Inhibition of ENO1 represents a viable therapeutic strategy for PAH. Data represent the mean ± SEM. Student t test and one-way ANOVA were used to compare two and multiple groups. Bonferroni post-tests were carried out after ANOVA
