Perillyle alcohol and Quercetin ameliorate monocrotaline-induced pulmonary artery hypertension in rats through PARP1-mediated miR-204 down-regulation and its downstream pathway

Figures

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  Fig. 1. Dose-response curve for determining the optimum dose of PA (a) and QS (b) to reduce pulmonary arteriole wall thickness (n = 5 in each group). For PA, dose of 50 mg/kg and for QS, dose of 30 mg/kg were selected for the main experiments. Values are mean ± SD. CTL, control; MCT, monocrotaline; Veh, vehicle; PA, perillyle alcohol; QS, quercetin

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  Fig. 2. Representative section of the lung of a normal rat (a), MCT-induced PAH (b), MCT + vehicle treatment (c), MCT + PA treatment (d) and MCT + QS treatment (e). f: relative quantitative analyses (mean ± SEM) of groups A-E. H&E Staining; magnification X 100. (n = 6 in each group). PA, Perillyle alcohol and QS, Quercetin. Scale bars are 20 μm. *** p < 0.001 vs control, ### p < 0.001 vs MCT + Veh

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  Fig. 3. Representative section of the lung of a normal rat (a), MCT induced PAH (b), MCT + vehicle treatment (c), MCT + PA treatment (d), and MCT + QS treatment (e), and (f) relative quantitative analyses (mean ± SEM) in groups A-E. Normal wall thickness is observed in A, and severe thickening is obvious in B and C. The lung of rats treated with PA and QS show mild thickening of arteriole wall. Here arteriole wall thickness in the CTL group is 14.3 μm, 44 μm in MCT, 43 μm in vehicle, and 18.2 μm and 15.3 μm in PA and QS, respectively. H&E staining; Magnification × 400. n = 6 in each group. Scale bars are 40 μm. *** p < 0.001 vs control, ### p < 0.001 vs MCT + Veh. CTL, control; MCT, monocrotaline; VE, vehicle; PA, perillyle alcohol; QS, quercetin

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  Fig. 4. Relative expression (Mean ± SEM) of α-SMA mRNA in the lungs of PAH rat models, and the effect of treatment with PA and QS. n = 6 in each group. *** and ### = P < 0.001, significantly different with both control and MCT + VE. CTL, control; MCT, monocrotaline; VE, vehicle; PA, perillyle alcohol; QS, quercetin; α-SMA, α-smooth muscle actin

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  Fig. 5. The expression of inflammatory cytokines IL-1β (a) and IL-8 (b) in monocrotaline-induced PAH lungs and the effect of treatment with PA and QS. n = 6 in each group. *** p < 0.001 vs Control, # p < 0.05, ### p < 0.001 vs MCT + Veh. CTL, control; MCT, monocrotaline; Veh, vehicle; PA, perillyle alcohol; QS, quercetin

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  Fig. 6. Relative expression (Mean ± SEM) of miR-204 (a), PARP1 mRNA (b) and PARP1 protein (c) in the lungs of studied groups. n = 6 in each group. The internal controls for PARP1 mRNA and miR-204 mRNA were 18 s rRNA and RNA U6 (RNU6), respectively. ** p < 0.01, *** p < 0.001 vs Control, ## p < 0.01, ### p < 0.001 vs MCT + Veh. CTL, control; MCT, monocrotaline; Veh, vehicle; PA, perillyle alcohol; QS, quercetin; PARP1, Poly ADP-ribose polymerase-1

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  Fig. 7. Relative expression (Mean ± SEM) of HIF1α and NFATc2 mRNA (a, c) and protein (b, d) in the lungs of PAH rat models, and the effect of treatment with perillyle alcohol and quercetin. n = 6 in each group. Internal control for HIF1α and NFATc2 mRNA was 18 s rRNA. *and # mean significantly different with control and MCT + Veh respectively. * p < 0.05, ** and ## p < 0.01, *** and ### p < 0.001. CTL, control; MCT, monocrotaline; Veh, vehicle; PA, perillyle alcohol; QS, quercetin; HIF1α, hypoxia-inducible factor 1-alpha; NFATc2, nuclear factor of activated T-cells, cytoplasmic 2