Integrating pharmacokinetics and network analysis to investigate the mechanism of Moutan Cortex in blood-heat and blood stasis syndrome

Figures

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  Fig. 1. Flow chart of integrating pharmacokinetics and network analysis to investigate the mechanism of Moutan Cortex in Blood-Heat and blood Stasis Syndrome

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  Fig. 2. Chemical structure of the analytes and IS

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  Fig. 3. UHPLC chromatograms of rats serum samples: A blank serum; B blank serum spiked with the nine analytes and IS; C serum sample of N-RMC at 1 h; D serum sample of BHS-RMC at 1 h; Standards; Standards: (1) 5-HMF; (2) Gallic acid; (3) Oxypaeoniflora; (4) Paeoniflorin; (5) 3, 8-Dihydroxy-2-methylchromone; (6) Benzoic acid; (7) Methyl paraben; (8) Paeonol; (9) Quercetin; (IS) 3-Hydroxy-4-methoxyacetophenone

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  Fig. 4.. 6-Keto-PGFlα (A), The TXB2 (B) and TXB2/6-Keto-PGFlα (C) value of rats in each group at the same time, “*” means there is a significant difference between the BHS model group and the Normal group (p < 0.05); “◆” means the BHS group is significantly different from the BHS-RMC group (p < 0.05); “▲” means BHS-RMC and Normal Significant difference between groups (p < 0.05)

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  Fig. 5. The mean plasma concentration time profiles of Gallic acid, 5-HMF, Oxypaeoniflorin, 3, 8-Dihydroxy-2-methylchromone, Paeoniflorin, Benzoic acid, Methylparaben, Quercetin and Paeonol after oral administration of RMC Extract (n = 6, mean ± SD)

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  Fig. 6. Network analysis of the nine compounds absorbed into the blood in treating BHS syndrome. A Targets attribution among the RMC and BHS syndrome. B Protein–protein interaction network of hub targets obtained from STRING database and constructed by Cytoscape, target gene indicated by green circle. C Top 20 KEGG pathway enrichment entries (p < 0.05). D Ingredients—target—pathway network. Labels Red diamonds, ingredients; green circles, protein targets; Blue V, Pathway; gray lines, compound–target pathway interactions. E Top 20 GO enrichment entries (p < 0.05)

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  Fig. 7. Binding compounds of RMC interacting with target proteins. A The scoring of binding force between 6 compounds and 8 proteins. B–G Binding studies of compounds–targets interactions. Molecules are represented by a yellow stick model, the hydrogen bonds are represented by a red dotted line. quercetin (B) paeoniflorin (C) within the F2 (3tu7) kinase active site respectively. paeonol (D), quercetin (E), Paeoniflorin (F), oxypaeoniflora (G) within the MAPK14 (ID = 1a9u) kinase active site respectively

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  Fig. 8. Production of F2 (A), F7 (B) and F10 (C) was determined by ELISA. Protein expression of F2 (D), F7 (E), F10 (F), AKT (G), MAPK14 (H) and MAPK10 (I) was determined by RT-qPCR. *p < 0.05, ** p < 0.01, *** p < 0.001, **** P < 0.0001 versus Model group