Effect of Apples on Hemolysis-Induced Hyperbilirubinemia: Evaluation by Network Pharmacology and Molecular Docking and Experimental Validation

Abstract

A network pharmacological approach was used to predict active ingredients in apples that may have therapeutic effects on hemolysis-induced hyperbilirubinemia and their potential targets, enrichment pathways and action process and to construct adisease-pathway-active component-target network for apples. Autodock software was used to perform molecular docking of key active ingredients and potential targets to verify the reliability of the network analysis results. Furthermore, the effect of apple alcohol extract (AE) on hemolysis-induced hyperbilirubinemia in rats was explored. The results showed that 17 active ingredients of apples with high gastrointestinal absorption and excellent drug-like properties were selected, among which, quercetin, 2α-hydroxyl-ursolic acid, β-sitosterol, phloretin and epicatechin could make important contribution to the treatment of hemolysis-induced hyperbilirubinemia. The top 5 targets in terms of moderate values in the network, including the serine/threonine protein kinase (Akt1), vascular endothelial growth factor A (VEGFA), epidermal growth factor receptor (EGFR), the tyrosine kinase (SRC), and signal transducer and activator of transcription 3 (STAT3) could exert therapeutic effects through the bile secretion pathway, the hypoxia inducible factor (HIF)-1 signaling pathway, the tumor necrosis factor (TNF) signaling pathway and the nuclear factor-kappa B (NF-κB) signaling pathway. Molecular docking was performed on the major active ingredients and the core targets selected by the network, and the results showed a strong affinity between the active ingredients and the potential targets with reliable results from the network analysis. The animal experimental results indicated that compared with the blank control group, the model group had a significant decrease in hemoglobin (HB) levels in the blood and a significant increase in the levels of total bilirubin (TB), indirect bilirubin (IB), alanine transaminase (ALT), aspartate transaminase (AST), TNF-α, and interleukin-6 (IL-6) (P < 0.01). Compared with the model group, HB levels increased and while the levels of TB, IB, ALT, AST, TNF-α and IL-6 significantly decreased in the AE intervention and positive control groups (P < 0.05 or P < 0.01). In summary, apples reduce hemolysis-induced hyperbilirubinemia, which will provide a basis for research on non-invasive therapy for neonatal jaundice.

Keywords

• apples; hemolysis-induced hyperbilirubinemia; network pharmacology; mechanism