Qiangli Wuhu mixture alleviates LPS-induced pneumonia by inhibiting the TLR4/NF-κB/NLRP3 pathway: a study based on network pharmacology

Abstract

Context: Qiangli Wuhu (QLWH) mixture is a concoction approved and registered by Ningxia Medical Products Administration. It has therapeutic effects on various types of pneumonia.

Objective: To clarify the mechanisms of QLWH in treating pneumonia.

Materials and methods: The potential targets of QLWH in the treatment of pneumonia were predicted by network pharmacology. Male, Institute of Cancer Research (ICR) mice were randomly divided into five groups of 12 mice, control, vehicle, QLWH (10 and 20 mg/kg) and dexamethasone (DXM), and orally treated twice daily with normal saline, QLWH or DXM. The pneumonia model was established by tracheal instillation of lipopolysaccharide (LPS). After treatment five days, ELISA, H&E staining and Western blot were used to investigate protective effects of QLWH.

Results: Nine hundred and ninety-four active ingredients were found through network pharmacology, corresponding to 135 targets for the treatment of pneumonia; compared to the vehicle group, QLWH (10 and 20 mg/kg) significantly decreased the levels of TNF-α (14.3% and 28.8%), IL-1β (23.9% and 42.8%) and IL-6 (13.2% and 16.1%), increased the levels of IL-10 (134.3% and 172.9%); in terms of mechanism, QLWH down-regulated TLR4/NF-κB/NLRP3 axis related proteins in lung tissue of pneumonia model mice (p < 0.05).

Discussion and conclusions: This study combined network pharmacology and animal experiments, providing effective evidence for the clinical promotion of QLWH. Meanwhile, it is of significance for further development.

Keywords: TLR signal pathway; Traditional Chinese medicine mixture; interaction network; pneumonia.

Figure 1.

Venn diagrams of therapeutic and pathogenic targets.

Figure 2.

Drug-component-target-disease interaction network diagram. Purple represents drugs, green represents 171 active components in Qiangli Wuhu mixture, blue represents 135 common targets and red represents diseases.

Figure 3.

Protein–protein interaction network. The colour and its shades represent the size of the degree value.

Figure 4.

Ranking of core targets based on topology analysis of the protein–protein interaction network.

Figure 5.

Gene Ontology enrichment analysis of Qiangli Wuhu mixture used to treat pneumonia. BP: biological processes; CC: cell component; MF: molecular function.

Figure 6.

Kyoto Encyclopaedia of Genes and Genomes enrichment analysis of Qiangli Wuhu mixture in the treatment of pneumonia. P adjust represents the significance of enrichment; the redder the colour, the higher the significance.

Figure 7.

Effects of Qiangli Wuhu mixture on serum levels of inflammatory cytokines TNF-α, IL-1β, IL-6 and anti-inflammatory cytokines IL-10 in mice with LPS-induced pneumonia. ^#p< 0.05 compared with the control group, *p< 0.05 compared with the vehicle group.

Figure 8.

Effect of Qiangli Wuhu mixture on lung tissue injury in mice with LPS-induced pneumonia. (A) Control group; (B) vehicle group; (C) low dose Qiangli Wuhu mixture (10 g/kg) group; (D) high dose Qiangli Wuhu mixture (20 g/kg) group; (E) DXM (10 mg/kg) group.

Figure 9.

Effect of Qiangli Wuhu mixture on NLRP3/TLR4/MyD88 pathway in mice with LPS-induced pneumonia. (A) Protein levels of NLRP3, MyD88 and TLR4 were detected by Western blot. (B–D) Protein levels of NF-kappa B, p-NF-kappa B, IκB-α, p-IκB-α, IKK-β and p-IKK-β were detected by Western blot. ^#p< 0.05 compared with the control group, *p< 0.05 compared with the vehicle group.