Comprehensive chemical analysis of polyphenols in the ethyl acetate extract from the roots of Ephedra sinica Stapf and evaluation of its therapeutic effects on SU5416/hypoxia-induced pulmonary arterial hypertension rats

Figures

• 

  Fig. 1. Schematic diagram of experimental design

• 

  Fig. 2. Tentative identification of proanthocyanidin monomers, dimers and trimers in the ethyl acetate extract of Ephedra roots

• 

  Fig. 3. Effects of ERE on pulmonary hemodynamics in SuHx rats. RVSP (A) and RVHI (B) in different groups of rats; C pulmonary artery acceleration time in different groups of rats; D right ventricular free wall thickness in different groups of rats; E representative images of right ventricular pressure waveforms and ultrasound results (F) in rats of each group. All the data were expressed as mean ± SD (n = 6). ###p < 0.001 compared with Control group; *p < 0.05, **p < 0.01, ***p < 0.001 compared with SuHx group. NS represented no statistical significance

• 

  Fig. 4. Effects of ERE on pulmonary vascular remodeling in SuHx rats. Representative images of H&E staining (A) and Masson staining (B) in the lung tissues of rats; The related indexes of pulmonary vascular remodeling, WT% (C) and WA% (D), were measured; E The percentage area of collagen within artery; F Representative images of immunohistochemical staining in the lung tissues of rats and integral optical density of α-SMA (G) in different groups; Levels of SOD (H) and MDA (I) in rat lung tissue. All the data were expressed as mean ± SD (n = 6). ###p < 0.001 compared with Control group; *p < 0.05, **p < 0.01, ***p < 0.001 compared with SuHx group. NS represented no statistical significance

• 

  Fig. 5. Effects of SuHx modeling and ERE intervention on serum metabolites in rats. A, B The metabolic profile for partial least squares discriminant analysis (PLS-DA) score scatter plots of serum samples (A: ESI ( +): R2X = 0.747, R2Y = 0.957, Q2 = 0.528; B: ESI (-): R2X = 0.596, R2Y = 0.832, Q2 = 0.528); C–F OPLS-DA models (Control vs. SuHx; SuHx vs. ERE-HD) for pairwise comparison

• 

  Fig. 6. The relative concentration levels of differential metabolites that related to the therapeutic effects of ERE on SU5416/hypoxia-induced PH rats. #p < 0.05, ##p < 0.01 compared with Control group; *p < 0.05, **p < 0.01, compared with SuHx group

• 

  Fig. 7. Effect of ERE on α and β diversity of intestinal microbiota in SuHx rats. The Chao 1 (A), Good’s Coverage (B), Shannon (C) and Simpson (D) indices of all samples (n = 5); Combined plot integrated with the PCA (E), PCoA (F), NMDS (G) and heatmaps (H) of the bacterial communities corresponding to beta diversity. All data are presented as the mean ± SD (n = 5)

• 

  Fig. 8. The effect of ERE on the gut microbiota of SU5416/hypoxia-induced PH rats. A Bacterial taxonomic characteristics of the intestinal microbiota at the phylum level; B The distribution map of intestinal microbiota with LDA scores exceeding 3 in the three groups of rats; C The relative abundance of bacterial communities in different groups at the family level; D The relative abundance of bacterial communities in different groups at the genus level

• 

  Fig. 9. A The relative abundance of differential intestinal microbial family of Muribaculaceae, Latobacillaceae, Peptostreptococcaceae, Desulfovibrionaceae, Clostridia_UCG-014 and Deferribacteraceae; B The relative abundance of differential intestinal microbial genus including Lactobacillus, Romboutsia, Alloprevotella and Colidextribacter in three groups; The p values were defined as follows: #P < 0.05, ##P < 0.01, ###P < 0.001(“#” represents a significant difference comparing with the Control group); *P < 0.05, **P < 0.01, ***P < 0.001 (“*” represents a significant difference comparing with the SuHx group)