Network pharmacology and molecular docking to explore the potential mechanism of chlorogenic acid in septic acute liver injury and experimental validation of TLR4/NF-κB pathway in vivo

Abstract

The objective of this study was to investigate the biological activities and mechanisms of chlorogenic acid (CGA) in the treatment of septic acute liver injury (SALI) based on the network pharmacology, molecular docking, in vivo studies, and other techniques. Chlorogenic acid and potential related targets of septic acute liver injury were searched from the public databases. Then, the protein-protein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted. Subsequently, molecular docking was performed to predict the binding of the active compound to the core target. Finally, in vivo experiments were carried out for further validation. A total of 60 common targets were identified between acute septic liver injury and chlorogenic acid, among which 9 common core targets (EGFR, ESR1, GSK3B, PTGS2, TLR4, PPARA, HSP90AA1, ACE, and MMP9) were screened with Cytoscape. Molecular docking indicated that these core targets had good binding activity to chlorogenic acid (- 7.2, - 6.8, - 7.7, - 8.7, - 6.1, - 6.8, - 7.3, - 8.4, and - 8.6 kcal/mol respectively). In the SALI mouse model, chlorogenic acid can improve pathological damage to the liver and apoptosis of liver cells, and anti-inflammatory properties significantly by the TLR4/NF-κB pathway (all P < 0.05). The biological activity and regulatory network of CGA on SALI were revealed, and the anti-inflammatory effect of CGA was verified, which could be associated with the TLR4/NF-κB pathway.

Keywords: Chlorogenic acid; Molecular docking; Network pharmacology; Septic acute liver injury; TLR4.

Fig. 1

Venn diagram of CGA and septic acute liver injury targets

Fig. 2

Network pharmacology diagram of CGA treatment for septic acute liver injury. A Targets intersection PPI network diagram between CGA and septic acute liver injury. B Topological analysis diagram of the targets intersection. C GO enrichment analysis diagram for targets intersection. D Analyzed targets intersection KEGG pathway map

Fig. 3

CGA docking with receptor protein molecules. CGA binds to A EGFR, B ESR1, C GSK3B, D PTGS2, E MMP9, F TLR4, G PPAR, H HSP90A1, I ACE, respectively

Fig. 4

Experimental validation of CGA treatment in a mouse model of SALI. A General liver of mice in each group. B HE staining of liver tissues in each group (Original magnification, 400 × . Scale bar = 40 μm); C pathological score of liver tissue in each group. D TUNNEL of liver cells in SALI mice. E The proportion of apoptotic cells n = 10, $\stackrel{-}{\mathrm{x}}$ ± s. ^###P < 0.001, ^***P < 0.001, ^###P < 0.001 vs the sham group; ^***P < 0.001 vs the CLP group

Fig. 5

Effect of CGA on inflammatory factors and TLR4/NF-κB pathway in SALI. A The expression of TNF-α in liver tissue of each group. B The expression of IL-1β in tissue of each group. C The Western blot strip of TLR4 and NF-κB (A represents sham group, B represents SALI group, C represents SALI + 20CGA group, D represents SALI + 100CGA group); D gray level analysis of TLR4 relative to αtubulin; E gray level analysis of NF-κB relative to αtubulin. n = 10. ###P < 0.001 vs the sham group; *P < 0.05, ***P < 0.001 vs the CGA group. ^#P < 0.05 vs the sham group; *P < 0.05 vs the CLP group. sham: sham group; CLP: cecum ligation and puncture