An integrated global chemomics and system biology approach to analyze the mechanisms of the traditional Chinese medicinal preparation Eriobotrya japonica - Fritillaria usuriensis dropping pills for pulmonary diseases

Abstract

Background: Traditional Chinese medicine (TCM) herbal formulae provide valuable therapeutic strategies. However, the active ingredients and mechanisms of action remain unclear for most of these formulae. Therefore, the identification of complex mechanisms is a major challenge in TCM research.

Methods: This study used a network pharmacology approach to clarify the anti-inflammatory and cough suppressing mechanisms of the Chinese medicinal preparation Eriobotrya japonica - Fritillaria usuriensis dropping pills (ChuanbeiPipa dropping pills, CBPP). The chemical constituents of CBPP were identified by high-quality ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS), and anti-inflammatory ingredients were selected and analyzed using the PharmMapper and Kyoto Encyclopedia of Genes and Genomes (KEGG) bioinformatics websites to predict the target proteins and related pathways, respectively. Then, an RNA-sequencing (RNA-Seq) analysis was carried out to investigate the different expression of genes in the lung tissue of rats with chronic bronchitis.

Results: Six main constituents affected 19 predicted pathways, including ursolic acid and oleanolic acid from Eriobotrya japonica (Thunb.) Lindl. (Eri), peiminine from Fritillaria usuriensis Maxim. (Fri), platycodigenin and polygalacic acid from Platycodon grandiflorum (Jacq.) A. DC. (Pla) and guanosine from Pinellia ternata (Thunb.) Makino. (Pin). Expression of 34 genes was significantly decreased after CBPP treatment, affecting four therapeutic functions: immunoregulation, anti-inflammation, collagen formation and muscle contraction.

Conclusion: The active components acted on the mitogen activated protein kinase (MAPK) pathway, transforming growth factor (TGF)-beta pathway, focal adhesion, tight junctions and the action cytoskeleton to exert anti-inflammatory effects, resolve phlegm, and relieve cough. This novel approach of global chemomics-integrated systems biology represents an effective and accurate strategy for the study of TCM with multiple components and multiple target mechanisms.

Fig. 1

UPLC/Q-TOF-MS analysis of CBPP extracts. a UPLC/UV chromatograms of the extracts; b, c TIC chromatograms in positive ESI mode and negative ESI mode, respectively

Fig. 2

Chemical structures of the CBPP extract components with anti-inflammatory activity

Fig. 3

CBPP alleviated LPS-induced chronic bronchitis in rats. a H&E staining images of rat lungs; b Leukocyte numbers in BALF and the ELISA of the inflammatory cytokines TNF-α and IL-8 in BALF and serum. ^* p < 0.05 and ^** p < 0.01 compared to the Con. ^# p < 0.05 and ^## p < 0.01, compared to the Mod (n = 10)

Fig. 4

Pathway prediction and RNA-Seq analysis of CBPP. a Structures of five representative active compounds that were identified by UPLC/Q-TOF-MS; b Main targets and pathways as analyzed by PharmMapper and KEGG, respectively (network analysis); c RNA-Seq-based transcriptome analysis by clustering and the functional classification of 34 up-regulated genes

Fig. 5

Network pharmacology analysis through the protein interaction of differentially expressed genes and protein targets and signaling pathway of representative ingredients of CBPP. The solid line represents an interaction directly confirmed through experiments or the literature, and the dotted line represents interactions indirectly speculated through docking or the literature