Mechanism of Eucommia ulmoides for pulmonary hypertension based on animal experiments and network pharmacology combined with molecular docking
Objective To determine the molecular mechanism of Eucommia ulmoides for pulmonary hypertension(PH)based on animal experiments and network pharmacology combined with molecular docking.Methods(1)The experimental PH rat model was established with a single intraperitoneal injection monocrotaline(MCT)and then the effect of extract from Eucommia ulmoides on PH rat was observed.Hematoxylin-eosin staining and enzyme linked immunosorbent assay were used to analyze the morphology of lung tissues and to detect HMGB1,TNF-α,IL-1β,and IL-6 level in the plasma.(2)Based on TCMSP,GeneCards,OMIM,TTD,and DrugBank,the target genes of Eucommia ulmoides and PH were screened and standardized.The Eucommia ulmoides ingredients-PH-target network was established with Cytoscape software.The key targets obtained were uploaded to the STRING database for protein-protein interaction(PPI)network analysis.The core targets of Eucommia ulmoides in the treatment of PH were identified by topological analysis.Additionally,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed.Molecular docking was used to verify the binding activity between the active ingredients of Eucommia ulmoides and the core target for PH.(3)Immunohistochemistry staining was used to detect the expression level of potential key targets.Results(1)Animal experiments show that the extract from Eucommia ulmoides relieved MCT-induced PH.Haemodynamic parameters including RVSP,RV/(LV+S),RV/(Tibial length)and the thickness of pulmonary arterial wall were greatly reduced in the PH rats.Meanwhile,the level of HMGB1,IL-1β,IL-6,and TNF-α in PH rats were reduced as well.(2)Network pharmacology showed 25 active components of Eucommia ulmoides,195 related targets,and 6416 PH-related targets.PPI network topology analysis uncovered 19 potential core targets of Eucommia ulmoides for PH treatment.A total of 2789 items were obtained via GO functional enrichment analysis,including 2477 biological processes,101 cellular components,and 211 molecular functions.Molecular docking showed that the active ingredients of Eucommia ulmoides had a strong binding affinity with the target protein.(3)Immunohistochemistry staining showed that Eucommia ulmoides obviously decreased the expression of AKT1,ESR1,STAT3 and IL-6,whereas increased the expression of TP53 in PH rats.Conclusion Eucommia ulmoides potentially contributes to the treatment of PH through multi-component,multi-target,and multi-pathway,providing a basis for its clinical application and further development.
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